Background Major depressive disorder(MDD),characterised by persistent anhedonia and elevated suicide risk,represents a global mental health challenge.Recent studies suggest a link between gut-brain axis dysfunction an...Background Major depressive disorder(MDD),characterised by persistent anhedonia and elevated suicide risk,represents a global mental health challenge.Recent studies suggest a link between gut-brain axis dysfunction and depression.The natural compound paeoniflorin demonstrates clinically relevant antidepressant effects,yet its underlying neurobiological mechanisms remain elusive.Aims This study aims to examine how paeoniflorin alleviates depression-like behaviours in rats subjected to chronic unpredictable mild stress(CUMS)by modulating the function of gut-brain axis,and explore the connections between gut microbiota,metabolites and MDD.Methods Depression-like behaviours in rats were induced by CUMS,and the antidepressant effect of paeoniflorin was assessed using behavioural tests.The composition and function of the intestinal microbiota were analysed using 16S rRNA sequencing,and metabolomic analysis was performed on serum,hippocampus,jejunum and faecal samples.Enzyme-linked immunosorbent assay and hematoxylin and eosin staining were used to detect the levels of inflammatory factors and cortisol,as well as the infiltration of inflammatory cells in the jejunum of rats after cohousing.Long-term potentiation assays and Golgi staining were used to detect dendritic spine density and synaptic plasticity,respectively.Results Paeoniflorin significantly alleviated depression-like behaviours and cognitive deficits in CUMS rats.16S rRNA sequencing revealed that paeoniflorin improved the abundance and diversity of the gut microbiota in CUMS rats.Enrichment of differential metabolites in the brain,intestine,faeces and serum revealed a primary accumulation in the amino acid metabolism pathway.We further observed a correlation between the relative abundance of microbial communities and metabolites.Cohousing experiments verified that microbial metabolites of paeoniflorin can reduce neuroinflammation and improve synaptic plasticity.Conclusions Disruptions in gut microbiota and its metabolites impair gut-brain interactions.Paeoniflorin’s neuroprotective and antidepressant effects are mediated through the modulation of the function of the gut-brain axis.展开更多
To evaluate the effect of components in Guanxin Ⅱ prescription on the pharmacokinetic profiles of paeoniflorin. Plasma concentration of Paeoniflorin in rats after intravenous injection of Paronia Pall Extract （PPE）...To evaluate the effect of components in Guanxin Ⅱ prescription on the pharmacokinetic profiles of paeoniflorin. Plasma concentration of Paeoniflorin in rats after intravenous injection of Paronia Pall Extract （PPE） and oral administration of PPE and three types of decoctions in Guanxin Ⅱ prescription, respectively, were determined by HPLC analyses. NONMEM （nonlinear mixed-effect modeling） method was used to analyze full set of pharmacokinetic data directly. A two-compartment model with first-order degradation in absorption compartment was employed for the data analysis. The mean of population parameters, CL1, V1, CL2, V2, Ka0, and Kal, were measured to be 0.509 L/h, 0.104 L, 0.113 L/h, 0.123 L, 0.135/h, and 0.0135/h, respectively. Inter-individual variabilities were estimated and dose formulation （DF） was identified as a significant covariate of Ka 1, Ka0, and V1. It is concluded that the pharmacokinetic behaviors of paeoniflorin in rats can alter with different dose formulations.展开更多
A simple, sensitive and specific high performance liquid chromatography-tandem mass spectrometry （HPLC-MS/MS）method combined with solid phase extraction has been developed and validated for the simultaneous quantifi...A simple, sensitive and specific high performance liquid chromatography-tandem mass spectrometry （HPLC-MS/MS）method combined with solid phase extraction has been developed and validated for the simultaneous quantification of paeoniflorinand albiflorin in rats plasma and tissue homogenate after administration of total glycosides of paeony （TGP）. Chromatographicseparation was achieved on a C15 column （150 mmx4.6 mm, 5 pro） with mobile phase consisted of acetonitrile-0.05% formicacid aqueous （20： 80, v/v） at a flow rate of 0.5 mL/min. The analytes were detected with triple-quadrupole mass spectrometer usingturbo ion spray with negative ionization in the multiple reaction-monitoring （MRM） modes. The results of method validationconformed to the requirements for the determination of biological samples. This method was successfully applied to thepharmacokinetics and tissue distribution study of TGP in rats. The results showed that paeonifiorin and albiflorin wereabsorbed and reached peak value quickly, and had the similar pharmacokinetic characteristics. Both of them underwent a rapid andwide distribution in the tissues throughout the whole body, among which stomach was the main distribution tissue. Moreover, theyhad the ability to cross the blood-brain barrier.展开更多
OBJECTIVE To establish a rapid,simple,precise and sensitive ultra performance liquid chromatography(UPLC) method for determination of paeoniflorin-6'-O-benzene sulfonate(acylated derivative of Pae,CP-25) and its p...OBJECTIVE To establish a rapid,simple,precise and sensitive ultra performance liquid chromatography(UPLC) method for determination of paeoniflorin-6'-O-benzene sulfonate(acylated derivative of Pae,CP-25) and its primary metabolite(Pae,M1) in rat plasma,and to investigate the effects of gender,food and disease status on the pharmacokinetics after oral administration in rats.METHODS Plasma samples and calibrators were extracted with methanol after addition of the internal standard solution.After evaporation of the methanol layer,the residue was reconstituted in mobile phase,and a 2 μL of the sample was injected into a Waters ACQUITY UPLC BEH C18(2.1 mm × 50 mm,1.7 μm) column for separation at a flow rate of 0.5 mL·min^(-1) at 40℃.The mobile phase was composed of 0.1% formic acid in water and methanol(68∶32,V/V).RESULTS The developed UPLC-MS/MS method was linear in the concentration range of 2-800 mg·L^(-1).Validation of the method proved that the method′s precision,selectivity and stability were all within the acceptable limits.Pharmacokinetics study showed that CP-25 could have more extensive distribution in females than that in males but no differences in M1.Food intake could also increase the extent of absorption and decrease the rate of clearance of CP-25 and M1 after oral administration in rats.The disease status would decrease the absorption of CP-25 in rats.Comparing single-and multiple-dosing in adjuvant arthritis(AA) model,absorption of CP-25 and M1 was improved with increasing dosage.CONCLUSION The developed UPLC-MS/MS method is sensitive,specific and was successfully applied to the pharmacokinetics of CP-25 and M1 in rats.And a significant gender,food intake and disease status differences for CP-25 and M1 were observed in this study.展开更多
Aim The present study aimed to investigate anti-inflammatory activity of 6-AP on murine model of aller- gic contact dermatitis （ACD）. Methods 6＇-acetylpaeoniflorin （6-AP） was synthesized from paeoniflorin （Pae）...Aim The present study aimed to investigate anti-inflammatory activity of 6-AP on murine model of aller- gic contact dermatitis （ACD）. Methods 6＇-acetylpaeoniflorin （6-AP） was synthesized from paeoniflorin （Pae） via acetylation and the structure was characterized by 1H-NMR, 13C-NMR and EI-MS. ACD model was established by repeated application of dinitrochlorobenzene （DNCB） to induce skin immune inflammation. The mice were oral- ly administered 6-AP （35, 70, 140 mg. kg-1 ·d^-l）, Pae （70 rag. kg-1·d^-1） and prednisone （Pre, 5 rag. kg^- 1· d^-1 ） from day 1 to day 7 after Cutaneous inflammation was evaluated by ear swelling and histological exami- nation. Splenocyte proliferation was assayed by the 3-（4, 5-dimethylthiazol-2-yl）-2, 5-diphenyl-2H tetrazolium bromide assay. The cytokine production in the splenocytes was measured by enzyme-linked immunosorbent assay. Results Topical application of DNCB to the skin provoked obvious swelling and inflammatory cell infiltration. 6- AP significantly inhibited ear swelling, decreased inflammatory cell infiltration and epidermal keratinization. Ad- dtionally, 6-AP obviously alleviated the hyperplasia of red pulp and germinal center （GC） appearance, decreased spleen index, decreased spleen index, and inhibited splenocyte proliferation in ACD model, compared to that of Pae. Further, the study indicated that 6-AP treatment could increase IL-10 level, while reduce IL-17 level in splenocytes simultaneously. The correlation analysis displayed significantly positive correlations between IL-17 level and the severity of skin inflammation, while negative correlations between IL-10 level and skin inflammation. Con- clusion 6-AP has a significantly higher anti-inflammatory effect than Pae, and it may be a useful treatment for ACD.展开更多
Aim Paeoniflorin-6＇O-benzene sulfonate （CP-25） was synthesized to improve oral absorption. This study was performed to investigate absorptive behavior and mechanism of CP-25 in intestine. Methods The effects of dru...Aim Paeoniflorin-6＇O-benzene sulfonate （CP-25） was synthesized to improve oral absorption. This study was performed to investigate absorptive behavior and mechanism of CP-25 in intestine. Methods The effects of drug concentration, intestinal segments, gender as well as ATP binding cassette （ABC） transporter inhibitors on absorption of CP-25 were studied in a situ single-pass intestinal perfusion rat model. Meanwhile, Paeoniflorin （Pae） was tested and served as control group. The concentration of tested drugs was measured by HPLC. Results The results showed intestinal absorption of CP-25 was neither segmental dependent changes nor gender difference. Transepithelial transportation would not change with increasing concentrations of CP-25, which suggest a passive transport was the main pattern of CP-25. Additionally, absorption of CP-25 was much better than that of Pae in small intestine. When compared with Pae, CP-25 gave a 1.82-fold permeability rate. Finally, the results indicated Pae was substrate of P-glycoprotein （P-gp） , but was not the substrate of breast cancer resistance protein and muhi- drug resistance associated protein 2. Among the used ABC inhibitors, the absorption rate of Pae could only be in- creased by？ P-gp inhibitor Verapamil and GF120918, while CP-25 had no remarkable alteration. Conclusion CP-25 has better absorptive features than that of Pae, which may be attributed to its lipophicity enhancement and be unaffected by P-gp efflux.展开更多
Alternatively activated macrophages are more frequently involved in tumor growth, angiogenesis, and immunosuppression. A previous study showed that paeoniflorin, the major active constituent of Paeonia lactiflora Pall...Alternatively activated macrophages are more frequently involved in tumor growth, angiogenesis, and immunosuppression. A previous study showed that paeoniflorin, the major active constituent of Paeonia lactiflora Pallas, can inhibit tumor growth and lung metastases of Lewis lung tumor-bearing mice. This study tried to investigate whether paeoniflorin inhibited lung cancer metastasis by inhibiting the alternative activation of macrophages(M2 macrophage). Using a viability assay, the cytotoxicity of paeoniflorin on Lewis lung cancer cells and peritoneal macrophages were investigated. In vitro scratch wound and in vivo lung metastasis experiments were used to test the ability to inhibit the migration of paeoniflorin and the function of M2 macrophages. Flow cytometry was performed to test the cell cycle of Lewis lung cancer cells, and to test the M2 macrophages in peritoneal macrophages and subcutaneous transplantable tumor. It was found that paeoniflorin showed no inhibitory effect on the growth of Lewis lung cancer cells and peritoneal macrophages of mouse in vitro. Paeoniflorin could attenuate the migration of LLC stimulated by alternatively activated macrophages(stimulated for 24 h and 48 h, paeoniflorin 1, 3, 10, 30, 100 μmol·L-1, P < 0.01 or P < 0.05 vs control group). Paeoniflorin could decrease the cell populations at S phases(paeoniflorin 10, 30, 100 μmol·L-1, P < 0.05 vs control group) and increase the cell populations at G0-G1 phases of Lewis lung cancer cells(paeoniflorin 100 μmol·L-1, P < 0.05 vs control group) and reduce the numbers of M2 macrophages in peritoneal macrophages induced by IL-4(paeoniflorin 1, 3, 10, 30, 100 μmol·L-1, P < 0.01 vs Control group). Paeoniflorin could reduce lung metastasis of Lewis lung cancer cells xenograft and decrease the numbers of M2 macrophages in subcutaneous xenograft tumour in vivo(paeoniflorin 20, 40 mg·kg-1, P < 0.01 vs control group). These results suggest that paeoniflorin could reduce lung metastasis of Lewis lung cancer cells xenograft partly through inhibiting the alternative activation of macrophages.展开更多
Paeonia lactiflora root(baishao in Chinese) is a commonly used herb in traditional Chinese medicines(TCM). Two isomers, paeoniflorin(PF) and albiflorin(AF), are isolated from P. lactiflora. The present study aimed to ...Paeonia lactiflora root(baishao in Chinese) is a commonly used herb in traditional Chinese medicines(TCM). Two isomers, paeoniflorin(PF) and albiflorin(AF), are isolated from P. lactiflora. The present study aimed to investigate the protective effects of PF and AF on myelosuppression induced by chemotherapy in mice and to explore the underlying mechanisms. The mouse myelosuppression model was established by intraperitoneal(i.p.) injection of cyclophosphamide(CP, 200 mg×kg^(–1)). The blood cell counts were performed. The thymus index and spleen index were also determined and bone morrow histological examination was performed. The levels of tumor necrosis factor-α(TNF-α) in serum and colony-stimulating factor(G-CSF) in plasma were measured by Enzyme-Linked Immunosorbent Assays(ELISA) and the serum levels of interleukin-3(IL-3), granulocyte-macrophagecolony- stimulatingfactor(GM-CSF), and interleukin-6(IL-6) were measured by radioimmunoassay(RIA). The levels of m RNA expression protein of IL-3, GM-CSF and G-CSF in spleen and bone marrow cells were determined respectively. PF and AF significantly increased the white blood cell(WBC) counts and reversed the atrophy of thymus. They also increased the serum levels of GM-CSF and IL-3 and the plasma level of G-CSF and reduced the level of TNF-α in serum.. PF enhanced the m RNA level of IL-3 and AF enhanced the m RNA levels of GM-CSF and G-CSF in the spleen. PF and AF both increased the protein levels of GM-CSF and G-CSF in bone marrow cells. In conclusion, our results demonstrated that PF and AF promoted the recovery of bone marrow hemopoietic function in the mouse myelosuppression model.展开更多
AIM: To examine the potential anti-tumor activity of paeoniflorin in the human gastric carcinoma cell line MGC-803.METHODS: Cell viability and cytotoxic effects in MGC-803 cells were analyzed using a 3-(4,5-dimethylth...AIM: To examine the potential anti-tumor activity of paeoniflorin in the human gastric carcinoma cell line MGC-803.METHODS: Cell viability and cytotoxic effects in MGC-803 cells were analyzed using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and lactate dehydrogenase assay, respectively. Cell apoptosis of MGC-803 cells was measured using flow cytometry,DAPI staining assay and caspase-3 activity assay.Quantitative reverse transcription-polymerase chain reaction(RT-PCR) was used to measure the expression of microRNA-124(miR-124) in response to paeoniflorin.The expression of phosphatidylinositol 3-kinase(PI3K),protein kinase B(Akt), phospho-Akt(p-Akt) and phospho-signal transducer and activator of transcription3(p-STAT3) were also measured by quantitative RTPCR and Western blot analysis in normal, miR-124 and anti-miR-124 over-expressing MGC-803 cells, treated with paeoniflorin.RESULTS: Paeoniflorin was found to inhibit MGC-803 cell viability in a dose-dependent manner. Paeoniflorin treatment was associated with the induction of apoptosis and caspase-3 activity in MGC-803 cells. Paeoniflorin treatment significantly increased miR-124 levels and inhibited the expression of PI3 K, Akt, p-Akt and p-STAT3 in MGC-803 cells. Interestingly, the over-expression of miR-124 inhibits PI3K/Akt and phospho-STAT3 expressions in MGC-803 cells. PI3 K agonist(IGF-1, 1μg/10 μL) or over-expression of STAT3 reversed the effect of paeoniflorin on the proliferation of MGC-803 cells. Over-expression of anti-miR-124 in MGC-803 cells reversed paeoniflorin-induced up-regulation.CONCLUSION: In summary, the in vitro data suggest that paeoniflorin is a potential novel therapeutic agent against gastric carcinoma, which inhibits cell viability and induces apoptosis through the up-regulation of miR-124 and suppression of PI3K/Akt and STAT3 signaling.展开更多
Paeoniflorin(PA) is an anti-Parkinson Chinese medicine with inferior bioavailability and difficulty in delivery to the brain. This research is to develop an efficacious PA nanocrystal formulation(PA-NCs) that is suita...Paeoniflorin(PA) is an anti-Parkinson Chinese medicine with inferior bioavailability and difficulty in delivery to the brain. This research is to develop an efficacious PA nanocrystal formulation(PA-NCs) that is suitable for intranasal administration to treat Parkinson’s disease(PD). PA-NCs were fabricated through an antisolvent precipitation method using TPGS as the stabilizer. The rod-shaped PA-NCs had particle size of 139.6 ± 1.3 nm and zeta potential of-23.2 ± 0.529 mV. A molecular dynamics simulation indicated that van der Waals forces are the primary drivers of interactions between PA and TPGS. In the ex vivo nasal mucosa permeation assay, the cumulative drug release at 24 h was 87.14% ± 5.34%,which was significantly higher than that of free PA. PA-NCs exhibited substantially improved cellular uptake as well as permeability on Calu-3 cells as compared to PA alone. FRET imaging analysis demonstrated that intact NCs could be internalized into Calu-3 cells.Moreover, PA-NCs conferred desirable protective effect against MPP+-induced SH-SY5Y cellular damage. Pharmacokinetic studies revealed a higher PA concentration in the brain following intranasal delivery of PA-NCs. In summary, the intranasal administration of PANCs is a promising treatment strategy for PD.展开更多
Depression ranks among the most common neuropsychiatric disorders globally.Current studies examining the roles of inflammation and mitochondrial autophagy in the antidepressant efficacy of paeoniflorin(PF)are sparse.T...Depression ranks among the most common neuropsychiatric disorders globally.Current studies examining the roles of inflammation and mitochondrial autophagy in the antidepressant efficacy of paeoniflorin(PF)are sparse.This study aimed to elucidate PF’s antidepressant mechanism by promoting autophagy and inhibiting NLRP3 inflammasome activation using chronic unpredictable mild stimulation(CUMS)-induced C57BL/6 mouse models in vivo and corticosterone(CORT)-induced HT22 cell models in vitro.Results demonstrated that PF enhanced the viability of HT22 cells following CORT exposure,restored mitochondrial membrane poten-tial(MMP),reduced reactive oxygen species accumulation,increased LC3 fluorescence intensity,and suppressed inflammatory cy-tokine secretion and inflammation activation.Additionally,PF ameliorated depressive behaviors induced by CUMS and improved damage in hippocampal neurons.It also reduced the expression of NLRP3,ASC,Caspase-1,IL-1β,and the assembly of the NLRP3 in-flammasome.Moreover,PF upregulated the expression of autophagy-related proteins in the hippocampus,facilitating the clearance of damaged mitochondria and enhancing autophagy.The role of autophagy in PF’s antidepressant effects was further confirmed through the use of the autophagy inhibitor 3-methyladenine(3-MA),which reduced the efficacy of PF.In conclusion,PF effectively improved depressive behaviors in CUMS-induced mice and reduced NLRP3-mediated inflammation both in vivo and in vitro,likely via the in-duction of autophagy.展开更多
OBJECTIVE: To investigate the sedative and hypnotic activity of paeoniflorin and freeze-dried Sini San powder on mice and provide a reliable method for determining the pharmacodynamic material basis of Sini San.METHOD...OBJECTIVE: To investigate the sedative and hypnotic activity of paeoniflorin and freeze-dried Sini San powder on mice and provide a reliable method for determining the pharmacodynamic material basis of Sini San.METHODS: Male adult mice weighing 20-22 g were used in this study. Three experiments were carried out. Synergism with pentobarbital was used as an index for hypnotic effect. Loss of the righting reflex was used to determine the start of sleep.Sleep latency and sleeping time were recorded in each experiment.RESULTS: The coefficient of variation of the suprathreshold dose(55 mg/kg) was significantly lower than that of the threshold dose. The sleep latency of mice was significantly decreased, and the sleeping time of mice was significantly prolonged. The effects of paeoniflorin and Sini San on prolonging the sleeping time of mice induced by pentobarbital sodium were significantly stronger than those in the control group.CONCLUSION: Paeoniflorin produces significant sedative and hypnotic effects, and there is an obvious dose-effect relationship.展开更多
Paeoniflorin (PF) is one of the main bioactive components of total glucosides of paeony (TGP) extracted from the root of Paeonia lactiflora Pall. TGP exhibit various biological activities such as improvement in memory...Paeoniflorin (PF) is one of the main bioactive components of total glucosides of paeony (TGP) extracted from the root of Paeonia lactiflora Pall. TGP exhibit various biological activities such as improvement in memory, hepatoprotection, antimutagenic properties and platelet aggregation inhibition. The aim of this paper is to review the pharmacokinetics (PK) of PF as a pure compound and in single or multiple herb(s) of traditional Chinese medicine (TCM) prescriptions. The distribution of PF or PF in TCM fitted one or two compartmental model after oral administration or intravenous injection, respectively. However, PF has a low bioavailability (BA) in rabbit (7.24%) and rat (3.24%) after oral administration. The PK profiles and BA of PF were remarkably improved when co-administered with sinomenine or glycyrrhizin acid. The PK profiles and BA of PF in Radix Paeonia Rubra (RP-R) and Jing-zhi guan-xin were improved, but in co-administration of RP-R with Radix Angelicae Sinensis, the BA was significantly reduced. PK profiles and BA of PF in Shan yao gan-cao tang or Danggui-Shaoyao-San was either remarkably improved or not. However, neither the PK profiles nor the BA of PF in Radix paeonia alba, Huangqin-tang Si ni san or Tang-Min-Ling-Wan was improved. Metabolism in the liver did not play any role in the low oral BA of PF. The low BA was thus attributed to poor permeation due to low lipophilicity, P-glycoprotein mediated efflux, intestinal bacteria and hydrolytic degradation in the intestine by the intestinal brush border lactase phlorizin hydrolase (LPH) and certain esterases. These findings show the in vivo course of PF and provide information on the maximum biological actions of PF that may help traditional Chinese herbal medicinal practitioners.展开更多
Objective:In this study,the influence of puerarin,paeoniflorin,and menthol on the structure and barrier function of tight junctions(TJs)in MadineDarby canine kidney epithelial(MDCK)and MDCK-multi-drug resistance 1(MDR...Objective:In this study,the influence of puerarin,paeoniflorin,and menthol on the structure and barrier function of tight junctions(TJs)in MadineDarby canine kidney epithelial(MDCK)and MDCK-multi-drug resistance 1(MDR1)cells was evaluated to determine the mechanisms by which the drugs cross the bloodebrain barrier(BBB).Method:Cells were treated with puerarin,paeoniflorin,and menthol followed by immunohistochemical staining with occludin,claudin-1,and F-actin.The cells were then observed using laser-scanning confocal microscopy.Average optical density(AOD)of the immunofluorescence images of the proteins were analyzed using ImageJ software while Transepithelial electrical resistance(TEER)was measured using an epithelial voltohmmeter.Results:Confocal microscopy revealed that puerarin-and paeoniflorin-treated tight junction proteins were conspicuous while menthol suppressed their expression.Correspondingly,AOD values of cells treated with puerarin or paeoniflorin,or both showed no difference compared to the control group(P>.05)while the menthol group value was downregulated.In 3 h,TEER of cells not treated with menthol were similar to the control group,while treatment with menthol significantly decreased TEER value(P<.05).In addition,application of menthol decreased TEER in MDCK cells earlier than in MDCK-MDR1 cells.Conclusion:Menthol but not puerarin and paeoniflorin may enhance paracellular transport and improve drug penetration of the BBB by disrupting the structure and,thereby,weakening the barrier function of TJs.展开更多
This study was to investigate the protective effect of paeoniflorin(PF)on hydrogen peroxide-induced injury.Firstly,“SMILES”of PF was searched in Pubchem and further was used for reverse molecular docking in Swiss Ta...This study was to investigate the protective effect of paeoniflorin(PF)on hydrogen peroxide-induced injury.Firstly,“SMILES”of PF was searched in Pubchem and further was used for reverse molecular docking in Swiss Target Prediction database to obtain potential targets.Injury-related molecules were obtained from GeenCards database,and the predicted targets of PF for injury treatment were selected by Wayne diagram.For mechanism analysis,the protein-protein interactions were constructed by String,and the KEGG analysis was conducted in Webgestalt.Then,cell viability and cytotoxicity assay were established by CCK8 assay.Also,the experimental cells were allocated to control,model(200μmol·L^(−1) H_(2)O_(2)),SB20358010μmol·L^(−1)(200μmol·L^(−1) H_(2)O_(2)+SB20358010μmol·L^(−1)),PF 50μmol·L^(−1)(200μmol·L^(−1) H_(2)O_(2)+PF 50μmol·L^(−1)),and PF 100μmol·L^(−1)(200μmol·L^(−1) H_(2)O_(2)+PF 100μmol·L^(−1))groups.We measured the intracellular ROS,Hoechst 33258 staining,cell apoptosis,the levels of Bcl-xl,Bcl-2,Caspase-3,Cleaved-caspase3,Cleaved-caspase7,TRPA1,TRPV1,and the phosphorylation expression of p38MAPK.There are 96 potential targets that may be associated with PF for injury treatment.Then,we chose the“Inflammatory mediator regulation of TRP channels”pathway for the experimental verification from the first 10 KEGG pathway.In experimental verification,H_(2)O_(2) decreased the cell viability moderately(P<0.05),and 100μmol·L^(−1) PF increased the cell viability significantly(P<0.05).Depending on the difference of intracellular ROS fluorescence intensity,PF inhibited H_(2)O_(2)-induced reactive oxygen species production in Schwann cells.In Hoechst 33258 staining,PF reversed the condensed chromatin and apoptotic nuclei following H_(2)O_(2) treatment.Moreover,Flow cytometry results showed that PF could substantially inhibit H_(2)O_(2) induced apoptosis(P<0.05).Pretreatment with PF obviously reduced the levels of Caspase3,Cleaved-caspase3,Cleaved-caspase7,TRPA1,TRPV1,and the phosphorylation expression of p38MAPK after H_(2)O_(2) treatment(P<0.05),increased the levels of Bcl-2,and Bcl-xl(P<0.05).PF inhibited Schwann cell injury and apoptosis induced by hydrogen peroxide,which mechanism was linked to the inhibition of phosphorylation of p38MAPK.展开更多
In order to evaluate the efficacy of traditional paeonia extract paeoniflorin against optic nerve crush, 16 Brown Norway rats were divided into two groups at random, with 8 rats in each group. In paeoniaflorin-treated...In order to evaluate the efficacy of traditional paeonia extract paeoniflorin against optic nerve crush, 16 Brown Norway rats were divided into two groups at random, with 8 rats in each group. In paeoniaflorin-treated group, 2 mg paeoniaflorin （total volum： 1 mL） was injected into rat＇s peritoneum one time a day for a period of 8 days. Rats in untreated group were given a single dose of vehicle. The optic nerve was crushed by a special forceps for 30 s in the left eye and a sham procedure was performed in the right eye on the 2nd day after the first injection. The retrograde fluorogold labeling of ganglion cells was conducted 5 days after optic nerve crush. The whole retina was flat-mounted thereafter. The ganglion cells that survived the crush were counted under fluorescent microscope by using an automatic counting software. As compared with the contralateral eye, the survival rate of ganglion cells in the left eye increased from 40.22% to 64.53% with a significant difference found between them （t=2.55, P=0.023）. The results suggested that the paeonia extract paeoniflorin possessed a protective effect against optic nerve crush.展开更多
Inhibiting the death receptor 3(DR3)signaling pathway in group 3 innate lymphoid cells(ILC3s)presents a promising approach for promoting mucosal repair in individuals with ulcerative colitis(UC).Paeoniflorin,a promine...Inhibiting the death receptor 3(DR3)signaling pathway in group 3 innate lymphoid cells(ILC3s)presents a promising approach for promoting mucosal repair in individuals with ulcerative colitis(UC).Paeoniflorin,a prominent component of Paeonia lactiflora Pall.,has demonstrated the ability to restore barrier function in UC mice,but the precise mechanism remains unclear.In this study,we aimed to delve into whether paeoniflorin may promote intestinal mucosal repair in chronic colitis by inhibiting DR3 signaling in ILC3s.C57BL/6 mice were subjected to random allocation into 7 distinct groups,namely the control group,the 2%dextran sodium sulfate(DSS)group,the paeoniflorin groups(25,50,and 100 mg/kg),the anti-tumor necrosis factor-like ligand 1A(anti-TL1A)antibody group,and the IgG group.We detected the expression of DR3 signaling pathway proteins and the proportion of ILC3s in the mouse colon using Western blot and flow cytometry,respectively.Meanwhile,DR3-overexpressing MNK-3 cells and 2%DSS-induced Rag1^(-/-)mice were used for verification.The results showed that paeoniflorin alleviated DSS-induced chronic colitis and repaired the intestinal mucosal barrier.Simultaneously,paeoniflorin inhibited the DR3 signaling pathway in ILC3s and regulated the content of cytokines(interleukin-17A,granulocyte-macrophage colony stimulating factor,and interleukin-22).Alternatively,paeoniflorin directly inhibited the DR3 signaling pathway in ILC3s to repair mucosal damage independently of the adaptive immune system.We additionally confirmed that paeoniflorin-conditioned medium(CM)restored the expression of tight junctions in Caco-2 cells via coculture.In conclusion,paeoniflorin ameliorates chronic colitis by enhancing the intestinal barrier in an ILC3-dependent manner,and its mechanism is associated with the inhibition of the DR3 signaling pathway.展开更多
A high sensitive method based on liquid chromatography tandem mass spectrometry（LC-MS/MS） was developed and validated for the study of permeability of danshensu（DS） and paeoniflorin（PF） in Caco-2 intestinal abso...A high sensitive method based on liquid chromatography tandem mass spectrometry（LC-MS/MS） was developed and validated for the study of permeability of danshensu（DS） and paeoniflorin（PF） in Caco-2 intestinal absorption model. The DS and PF were extracted from cell culture by vacuum-lyophilizing and then separated on a Zorbax Stable Bond C18 column with 0.1% acetic acid aqueous solution and methanol as mobile phase. Detection was carried out by negative electrospray ionization（ESI ） with selected reaction monitoring（SRM） mode. The apparent permeability coefficients（Papp） of DS and PF in Caco-2 cell medium were calculated and the effects of verapamil on the coefficients Papp of the two test compounds were also illustrated. The permeability of PF was much better than that of DS when the two compounds were administrated individually. Co-administration of DS and PF led to the decrease of the transport from apical side to basolateral side for both the compounds. However, the transport in the contrary direction were accelerated. It was also observed that verapamil could accelerate the transport of the test compounds from apical side to basolateral side. However, the absorption-enhanced effect of verapamil was attenuated when DS and PF were co-administrated. These observations suggest that both passive diffusion and active efflux involved in P-gp would effect the passage of DS and PF across Caco-2 cell monolayer. At the same time, the co-administration of DS and PF to an alteration of transport behavior, which suggests that the interaction must be taken into account when ‘n-in-one＇ samples were used in Caco-2 intestinal model.展开更多
In order to facilitate the preparation of paeoniflorin(PF)and albiflorin(AF),two chief bioactive constituents in Paeonia lactiflora Pall(PL),induction and culture of callus from PL were studied.With a modified woody p...In order to facilitate the preparation of paeoniflorin(PF)and albiflorin(AF),two chief bioactive constituents in Paeonia lactiflora Pall(PL),induction and culture of callus from PL were studied.With a modified woody plant medium supplemented with 0.5 mg·L-16-benzylaminopurine,1.0 mg·L-1naphthylacetic acid,0.1 mg·L-1thidiazuron and 30 g·L-1sucrose,callus was induced from four kinds of explants:leaf,stems,petiole,and root.The potency to form callus varies between different explants and leaf explants exhibits the highest capacity(100%).On the other hand,root-derived callus(R-callus)produces the highest level of total amount of PF and AF,31.8 mg·g-1dry mass,which is higher than the corresponding level in the root of field cultivated PL.Furthermore,the time needed is only 40 days,remarkably shorter than the cultivation time of PL,about 4–5 years.Higher accumulation levels of PF and AF with shorter production time indicate that callus culture of PL is a promising powerful tool for production of PF and AF in the future.展开更多
Insulin-like growth factor-1(IGF-1)is considered as a pathogenic factor contributing to sebaceous gland dysfunction,which leads to acne vulgaris.Paeoniflorin(Pae),a bioactive monomer derived from total glycosides of p...Insulin-like growth factor-1(IGF-1)is considered as a pathogenic factor contributing to sebaceous gland dysfunction,which leads to acne vulgaris.Paeoniflorin(Pae),a bioactive monomer derived from total glycosides of paeony,has shown potential in treating various diseases.However,its anti-acne effects on human sebocytes are not well understood.In this study,we investigated the effects of Pae on acne development induced by IGF-1 in SZ95 sebocytes.Following IGF-1 stimulation,SZ95 sebocytes were exposed to Pae and then determined for proliferation,cell cycle,apoptosis,lipogenesis and pro-inflammatory cytokine secretion.We also analyzed the expression of proteins involved in the PI3K/Akt/FoxO1 and JAK2/STAT3 pathways.In vitro experiments demonstrated that Pae significantly inhibited colony formation,induced G1/S cell cycle arrest,promoted apoptosis,inhibited lipogenesis and cytokine synthesis in IGF-1-treated SZ95 sebocytes.Furthermore,Pae suppressed the phosphorylation of Akt,FoxO1,JAK2,and STAT3.Importantly,the sebo-suppressive and anti-inflammatory effects of Pae were enhanced by blocking PI3K and JAK2.In summary,our findings suggest that Pae has potent anti-proliferative and pro-apoptotic effects in SZ95 sebocytes.Additionally,Pae effectively protects against IGF-1-induced lipogenesis and inflammation by targeting the PI3K/Akt/FoxO1 and JAK2/STAT3 signaling pathways.展开更多
基金supported by the National Natural Science Foundation of China(No 82305144)the Jiangsu Provincial Natural Science Foundation Project for Universities(No 23KJB360004)the National Natural Science Foundation Supporting Project of Nanjing University of Chinese Medicine(No XPT82305144).
文摘Background Major depressive disorder(MDD),characterised by persistent anhedonia and elevated suicide risk,represents a global mental health challenge.Recent studies suggest a link between gut-brain axis dysfunction and depression.The natural compound paeoniflorin demonstrates clinically relevant antidepressant effects,yet its underlying neurobiological mechanisms remain elusive.Aims This study aims to examine how paeoniflorin alleviates depression-like behaviours in rats subjected to chronic unpredictable mild stress(CUMS)by modulating the function of gut-brain axis,and explore the connections between gut microbiota,metabolites and MDD.Methods Depression-like behaviours in rats were induced by CUMS,and the antidepressant effect of paeoniflorin was assessed using behavioural tests.The composition and function of the intestinal microbiota were analysed using 16S rRNA sequencing,and metabolomic analysis was performed on serum,hippocampus,jejunum and faecal samples.Enzyme-linked immunosorbent assay and hematoxylin and eosin staining were used to detect the levels of inflammatory factors and cortisol,as well as the infiltration of inflammatory cells in the jejunum of rats after cohousing.Long-term potentiation assays and Golgi staining were used to detect dendritic spine density and synaptic plasticity,respectively.Results Paeoniflorin significantly alleviated depression-like behaviours and cognitive deficits in CUMS rats.16S rRNA sequencing revealed that paeoniflorin improved the abundance and diversity of the gut microbiota in CUMS rats.Enrichment of differential metabolites in the brain,intestine,faeces and serum revealed a primary accumulation in the amino acid metabolism pathway.We further observed a correlation between the relative abundance of microbial communities and metabolites.Cohousing experiments verified that microbial metabolites of paeoniflorin can reduce neuroinflammation and improve synaptic plasticity.Conclusions Disruptions in gut microbiota and its metabolites impair gut-brain interactions.Paeoniflorin’s neuroprotective and antidepressant effects are mediated through the modulation of the function of the gut-brain axis.
基金National Natural Science Foundation (Grant No. 30472165)
文摘To evaluate the effect of components in Guanxin Ⅱ prescription on the pharmacokinetic profiles of paeoniflorin. Plasma concentration of Paeoniflorin in rats after intravenous injection of Paronia Pall Extract （PPE） and oral administration of PPE and three types of decoctions in Guanxin Ⅱ prescription, respectively, were determined by HPLC analyses. NONMEM （nonlinear mixed-effect modeling） method was used to analyze full set of pharmacokinetic data directly. A two-compartment model with first-order degradation in absorption compartment was employed for the data analysis. The mean of population parameters, CL1, V1, CL2, V2, Ka0, and Kal, were measured to be 0.509 L/h, 0.104 L, 0.113 L/h, 0.123 L, 0.135/h, and 0.0135/h, respectively. Inter-individual variabilities were estimated and dose formulation （DF） was identified as a significant covariate of Ka 1, Ka0, and V1. It is concluded that the pharmacokinetic behaviors of paeoniflorin in rats can alter with different dose formulations.
基金Natural Science Foundation of Hebei Province of China(Grant No.C2011206158,08B031)Hundreds of Innovative Talents Project of Hebei Education Department of China
文摘A simple, sensitive and specific high performance liquid chromatography-tandem mass spectrometry （HPLC-MS/MS）method combined with solid phase extraction has been developed and validated for the simultaneous quantification of paeoniflorinand albiflorin in rats plasma and tissue homogenate after administration of total glycosides of paeony （TGP）. Chromatographicseparation was achieved on a C15 column （150 mmx4.6 mm, 5 pro） with mobile phase consisted of acetonitrile-0.05% formicacid aqueous （20： 80, v/v） at a flow rate of 0.5 mL/min. The analytes were detected with triple-quadrupole mass spectrometer usingturbo ion spray with negative ionization in the multiple reaction-monitoring （MRM） modes. The results of method validationconformed to the requirements for the determination of biological samples. This method was successfully applied to thepharmacokinetics and tissue distribution study of TGP in rats. The results showed that paeonifiorin and albiflorin wereabsorbed and reached peak value quickly, and had the similar pharmacokinetic characteristics. Both of them underwent a rapid andwide distribution in the tissues throughout the whole body, among which stomach was the main distribution tissue. Moreover, theyhad the ability to cross the blood-brain barrier.
基金supported by National Natural Science Foundation of China(81330081 and 81302845)
文摘OBJECTIVE To establish a rapid,simple,precise and sensitive ultra performance liquid chromatography(UPLC) method for determination of paeoniflorin-6'-O-benzene sulfonate(acylated derivative of Pae,CP-25) and its primary metabolite(Pae,M1) in rat plasma,and to investigate the effects of gender,food and disease status on the pharmacokinetics after oral administration in rats.METHODS Plasma samples and calibrators were extracted with methanol after addition of the internal standard solution.After evaporation of the methanol layer,the residue was reconstituted in mobile phase,and a 2 μL of the sample was injected into a Waters ACQUITY UPLC BEH C18(2.1 mm × 50 mm,1.7 μm) column for separation at a flow rate of 0.5 mL·min^(-1) at 40℃.The mobile phase was composed of 0.1% formic acid in water and methanol(68∶32,V/V).RESULTS The developed UPLC-MS/MS method was linear in the concentration range of 2-800 mg·L^(-1).Validation of the method proved that the method′s precision,selectivity and stability were all within the acceptable limits.Pharmacokinetics study showed that CP-25 could have more extensive distribution in females than that in males but no differences in M1.Food intake could also increase the extent of absorption and decrease the rate of clearance of CP-25 and M1 after oral administration in rats.The disease status would decrease the absorption of CP-25 in rats.Comparing single-and multiple-dosing in adjuvant arthritis(AA) model,absorption of CP-25 and M1 was improved with increasing dosage.CONCLUSION The developed UPLC-MS/MS method is sensitive,specific and was successfully applied to the pharmacokinetics of CP-25 and M1 in rats.And a significant gender,food intake and disease status differences for CP-25 and M1 were observed in this study.
文摘Aim The present study aimed to investigate anti-inflammatory activity of 6-AP on murine model of aller- gic contact dermatitis （ACD）. Methods 6＇-acetylpaeoniflorin （6-AP） was synthesized from paeoniflorin （Pae） via acetylation and the structure was characterized by 1H-NMR, 13C-NMR and EI-MS. ACD model was established by repeated application of dinitrochlorobenzene （DNCB） to induce skin immune inflammation. The mice were oral- ly administered 6-AP （35, 70, 140 mg. kg-1 ·d^-l）, Pae （70 rag. kg-1·d^-1） and prednisone （Pre, 5 rag. kg^- 1· d^-1 ） from day 1 to day 7 after Cutaneous inflammation was evaluated by ear swelling and histological exami- nation. Splenocyte proliferation was assayed by the 3-（4, 5-dimethylthiazol-2-yl）-2, 5-diphenyl-2H tetrazolium bromide assay. The cytokine production in the splenocytes was measured by enzyme-linked immunosorbent assay. Results Topical application of DNCB to the skin provoked obvious swelling and inflammatory cell infiltration. 6- AP significantly inhibited ear swelling, decreased inflammatory cell infiltration and epidermal keratinization. Ad- dtionally, 6-AP obviously alleviated the hyperplasia of red pulp and germinal center （GC） appearance, decreased spleen index, decreased spleen index, and inhibited splenocyte proliferation in ACD model, compared to that of Pae. Further, the study indicated that 6-AP treatment could increase IL-10 level, while reduce IL-17 level in splenocytes simultaneously. The correlation analysis displayed significantly positive correlations between IL-17 level and the severity of skin inflammation, while negative correlations between IL-10 level and skin inflammation. Con- clusion 6-AP has a significantly higher anti-inflammatory effect than Pae, and it may be a useful treatment for ACD.
文摘Aim Paeoniflorin-6＇O-benzene sulfonate （CP-25） was synthesized to improve oral absorption. This study was performed to investigate absorptive behavior and mechanism of CP-25 in intestine. Methods The effects of drug concentration, intestinal segments, gender as well as ATP binding cassette （ABC） transporter inhibitors on absorption of CP-25 were studied in a situ single-pass intestinal perfusion rat model. Meanwhile, Paeoniflorin （Pae） was tested and served as control group. The concentration of tested drugs was measured by HPLC. Results The results showed intestinal absorption of CP-25 was neither segmental dependent changes nor gender difference. Transepithelial transportation would not change with increasing concentrations of CP-25, which suggest a passive transport was the main pattern of CP-25. Additionally, absorption of CP-25 was much better than that of Pae in small intestine. When compared with Pae, CP-25 gave a 1.82-fold permeability rate. Finally, the results indicated Pae was substrate of P-glycoprotein （P-gp） , but was not the substrate of breast cancer resistance protein and muhi- drug resistance associated protein 2. Among the used ABC inhibitors, the absorption rate of Pae could only be in- creased by？ P-gp inhibitor Verapamil and GF120918, while CP-25 had no remarkable alteration. Conclusion CP-25 has better absorptive features than that of Pae, which may be attributed to its lipophicity enhancement and be unaffected by P-gp efflux.
基金supported by National Natural Science Foundation of China(No.81503284)Fundamental Research Funds for the Central Universities(No.2015PY016)Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD)
文摘Alternatively activated macrophages are more frequently involved in tumor growth, angiogenesis, and immunosuppression. A previous study showed that paeoniflorin, the major active constituent of Paeonia lactiflora Pallas, can inhibit tumor growth and lung metastases of Lewis lung tumor-bearing mice. This study tried to investigate whether paeoniflorin inhibited lung cancer metastasis by inhibiting the alternative activation of macrophages(M2 macrophage). Using a viability assay, the cytotoxicity of paeoniflorin on Lewis lung cancer cells and peritoneal macrophages were investigated. In vitro scratch wound and in vivo lung metastasis experiments were used to test the ability to inhibit the migration of paeoniflorin and the function of M2 macrophages. Flow cytometry was performed to test the cell cycle of Lewis lung cancer cells, and to test the M2 macrophages in peritoneal macrophages and subcutaneous transplantable tumor. It was found that paeoniflorin showed no inhibitory effect on the growth of Lewis lung cancer cells and peritoneal macrophages of mouse in vitro. Paeoniflorin could attenuate the migration of LLC stimulated by alternatively activated macrophages(stimulated for 24 h and 48 h, paeoniflorin 1, 3, 10, 30, 100 μmol·L-1, P < 0.01 or P < 0.05 vs control group). Paeoniflorin could decrease the cell populations at S phases(paeoniflorin 10, 30, 100 μmol·L-1, P < 0.05 vs control group) and increase the cell populations at G0-G1 phases of Lewis lung cancer cells(paeoniflorin 100 μmol·L-1, P < 0.05 vs control group) and reduce the numbers of M2 macrophages in peritoneal macrophages induced by IL-4(paeoniflorin 1, 3, 10, 30, 100 μmol·L-1, P < 0.01 vs Control group). Paeoniflorin could reduce lung metastasis of Lewis lung cancer cells xenograft and decrease the numbers of M2 macrophages in subcutaneous xenograft tumour in vivo(paeoniflorin 20, 40 mg·kg-1, P < 0.01 vs control group). These results suggest that paeoniflorin could reduce lung metastasis of Lewis lung cancer cells xenograft partly through inhibiting the alternative activation of macrophages.
基金supported by the National Nature Science Foundation of China(Nos.81473370,81173569)
文摘Paeonia lactiflora root(baishao in Chinese) is a commonly used herb in traditional Chinese medicines(TCM). Two isomers, paeoniflorin(PF) and albiflorin(AF), are isolated from P. lactiflora. The present study aimed to investigate the protective effects of PF and AF on myelosuppression induced by chemotherapy in mice and to explore the underlying mechanisms. The mouse myelosuppression model was established by intraperitoneal(i.p.) injection of cyclophosphamide(CP, 200 mg×kg^(–1)). The blood cell counts were performed. The thymus index and spleen index were also determined and bone morrow histological examination was performed. The levels of tumor necrosis factor-α(TNF-α) in serum and colony-stimulating factor(G-CSF) in plasma were measured by Enzyme-Linked Immunosorbent Assays(ELISA) and the serum levels of interleukin-3(IL-3), granulocyte-macrophagecolony- stimulatingfactor(GM-CSF), and interleukin-6(IL-6) were measured by radioimmunoassay(RIA). The levels of m RNA expression protein of IL-3, GM-CSF and G-CSF in spleen and bone marrow cells were determined respectively. PF and AF significantly increased the white blood cell(WBC) counts and reversed the atrophy of thymus. They also increased the serum levels of GM-CSF and IL-3 and the plasma level of G-CSF and reduced the level of TNF-α in serum.. PF enhanced the m RNA level of IL-3 and AF enhanced the m RNA levels of GM-CSF and G-CSF in the spleen. PF and AF both increased the protein levels of GM-CSF and G-CSF in bone marrow cells. In conclusion, our results demonstrated that PF and AF promoted the recovery of bone marrow hemopoietic function in the mouse myelosuppression model.
基金Supported by National Natural Science Foundation of China,No.81372553
文摘AIM: To examine the potential anti-tumor activity of paeoniflorin in the human gastric carcinoma cell line MGC-803.METHODS: Cell viability and cytotoxic effects in MGC-803 cells were analyzed using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and lactate dehydrogenase assay, respectively. Cell apoptosis of MGC-803 cells was measured using flow cytometry,DAPI staining assay and caspase-3 activity assay.Quantitative reverse transcription-polymerase chain reaction(RT-PCR) was used to measure the expression of microRNA-124(miR-124) in response to paeoniflorin.The expression of phosphatidylinositol 3-kinase(PI3K),protein kinase B(Akt), phospho-Akt(p-Akt) and phospho-signal transducer and activator of transcription3(p-STAT3) were also measured by quantitative RTPCR and Western blot analysis in normal, miR-124 and anti-miR-124 over-expressing MGC-803 cells, treated with paeoniflorin.RESULTS: Paeoniflorin was found to inhibit MGC-803 cell viability in a dose-dependent manner. Paeoniflorin treatment was associated with the induction of apoptosis and caspase-3 activity in MGC-803 cells. Paeoniflorin treatment significantly increased miR-124 levels and inhibited the expression of PI3 K, Akt, p-Akt and p-STAT3 in MGC-803 cells. Interestingly, the over-expression of miR-124 inhibits PI3K/Akt and phospho-STAT3 expressions in MGC-803 cells. PI3 K agonist(IGF-1, 1μg/10 μL) or over-expression of STAT3 reversed the effect of paeoniflorin on the proliferation of MGC-803 cells. Over-expression of anti-miR-124 in MGC-803 cells reversed paeoniflorin-induced up-regulation.CONCLUSION: In summary, the in vitro data suggest that paeoniflorin is a potential novel therapeutic agent against gastric carcinoma, which inhibits cell viability and induces apoptosis through the up-regulation of miR-124 and suppression of PI3K/Akt and STAT3 signaling.
基金the Guangdong Provincial Natural Science Foundation of China(2018A030310623)the Guangdong Provincial Medical Scientific Research Foundation of China(A2019027)the Guangzhou Science Technology and Innovation Commission Technology Research Projects(201805010005)。
文摘Paeoniflorin(PA) is an anti-Parkinson Chinese medicine with inferior bioavailability and difficulty in delivery to the brain. This research is to develop an efficacious PA nanocrystal formulation(PA-NCs) that is suitable for intranasal administration to treat Parkinson’s disease(PD). PA-NCs were fabricated through an antisolvent precipitation method using TPGS as the stabilizer. The rod-shaped PA-NCs had particle size of 139.6 ± 1.3 nm and zeta potential of-23.2 ± 0.529 mV. A molecular dynamics simulation indicated that van der Waals forces are the primary drivers of interactions between PA and TPGS. In the ex vivo nasal mucosa permeation assay, the cumulative drug release at 24 h was 87.14% ± 5.34%,which was significantly higher than that of free PA. PA-NCs exhibited substantially improved cellular uptake as well as permeability on Calu-3 cells as compared to PA alone. FRET imaging analysis demonstrated that intact NCs could be internalized into Calu-3 cells.Moreover, PA-NCs conferred desirable protective effect against MPP+-induced SH-SY5Y cellular damage. Pharmacokinetic studies revealed a higher PA concentration in the brain following intranasal delivery of PA-NCs. In summary, the intranasal administration of PANCs is a promising treatment strategy for PD.
基金supported by the Jilin Science&Technology Development Plan(Nos.20210204001YY,20200404090YY)Jilin Province aid Project for Xinjiang Uygur Autonomous Region(No.0207-202020043).
文摘Depression ranks among the most common neuropsychiatric disorders globally.Current studies examining the roles of inflammation and mitochondrial autophagy in the antidepressant efficacy of paeoniflorin(PF)are sparse.This study aimed to elucidate PF’s antidepressant mechanism by promoting autophagy and inhibiting NLRP3 inflammasome activation using chronic unpredictable mild stimulation(CUMS)-induced C57BL/6 mouse models in vivo and corticosterone(CORT)-induced HT22 cell models in vitro.Results demonstrated that PF enhanced the viability of HT22 cells following CORT exposure,restored mitochondrial membrane poten-tial(MMP),reduced reactive oxygen species accumulation,increased LC3 fluorescence intensity,and suppressed inflammatory cy-tokine secretion and inflammation activation.Additionally,PF ameliorated depressive behaviors induced by CUMS and improved damage in hippocampal neurons.It also reduced the expression of NLRP3,ASC,Caspase-1,IL-1β,and the assembly of the NLRP3 in-flammasome.Moreover,PF upregulated the expression of autophagy-related proteins in the hippocampus,facilitating the clearance of damaged mitochondria and enhancing autophagy.The role of autophagy in PF’s antidepressant effects was further confirmed through the use of the autophagy inhibitor 3-methyladenine(3-MA),which reduced the efficacy of PF.In conclusion,PF effectively improved depressive behaviors in CUMS-induced mice and reduced NLRP3-mediated inflammation both in vivo and in vitro,likely via the in-duction of autophagy.
基金Supported by Gansu Province Finance colleges Fundamental Research Foundation (No.2013-2)Ministry of Education,Science and Technology Key Project (No.212186)and Gansu Province Natural Science Foundation (No.1010RJZA212)
文摘OBJECTIVE: To investigate the sedative and hypnotic activity of paeoniflorin and freeze-dried Sini San powder on mice and provide a reliable method for determining the pharmacodynamic material basis of Sini San.METHODS: Male adult mice weighing 20-22 g were used in this study. Three experiments were carried out. Synergism with pentobarbital was used as an index for hypnotic effect. Loss of the righting reflex was used to determine the start of sleep.Sleep latency and sleeping time were recorded in each experiment.RESULTS: The coefficient of variation of the suprathreshold dose(55 mg/kg) was significantly lower than that of the threshold dose. The sleep latency of mice was significantly decreased, and the sleeping time of mice was significantly prolonged. The effects of paeoniflorin and Sini San on prolonging the sleeping time of mice induced by pentobarbital sodium were significantly stronger than those in the control group.CONCLUSION: Paeoniflorin produces significant sedative and hypnotic effects, and there is an obvious dose-effect relationship.
文摘Paeoniflorin (PF) is one of the main bioactive components of total glucosides of paeony (TGP) extracted from the root of Paeonia lactiflora Pall. TGP exhibit various biological activities such as improvement in memory, hepatoprotection, antimutagenic properties and platelet aggregation inhibition. The aim of this paper is to review the pharmacokinetics (PK) of PF as a pure compound and in single or multiple herb(s) of traditional Chinese medicine (TCM) prescriptions. The distribution of PF or PF in TCM fitted one or two compartmental model after oral administration or intravenous injection, respectively. However, PF has a low bioavailability (BA) in rabbit (7.24%) and rat (3.24%) after oral administration. The PK profiles and BA of PF were remarkably improved when co-administered with sinomenine or glycyrrhizin acid. The PK profiles and BA of PF in Radix Paeonia Rubra (RP-R) and Jing-zhi guan-xin were improved, but in co-administration of RP-R with Radix Angelicae Sinensis, the BA was significantly reduced. PK profiles and BA of PF in Shan yao gan-cao tang or Danggui-Shaoyao-San was either remarkably improved or not. However, neither the PK profiles nor the BA of PF in Radix paeonia alba, Huangqin-tang Si ni san or Tang-Min-Ling-Wan was improved. Metabolism in the liver did not play any role in the low oral BA of PF. The low BA was thus attributed to poor permeation due to low lipophilicity, P-glycoprotein mediated efflux, intestinal bacteria and hydrolytic degradation in the intestine by the intestinal brush border lactase phlorizin hydrolase (LPH) and certain esterases. These findings show the in vivo course of PF and provide information on the maximum biological actions of PF that may help traditional Chinese herbal medicinal practitioners.
文摘Objective:In this study,the influence of puerarin,paeoniflorin,and menthol on the structure and barrier function of tight junctions(TJs)in MadineDarby canine kidney epithelial(MDCK)and MDCK-multi-drug resistance 1(MDR1)cells was evaluated to determine the mechanisms by which the drugs cross the bloodebrain barrier(BBB).Method:Cells were treated with puerarin,paeoniflorin,and menthol followed by immunohistochemical staining with occludin,claudin-1,and F-actin.The cells were then observed using laser-scanning confocal microscopy.Average optical density(AOD)of the immunofluorescence images of the proteins were analyzed using ImageJ software while Transepithelial electrical resistance(TEER)was measured using an epithelial voltohmmeter.Results:Confocal microscopy revealed that puerarin-and paeoniflorin-treated tight junction proteins were conspicuous while menthol suppressed their expression.Correspondingly,AOD values of cells treated with puerarin or paeoniflorin,or both showed no difference compared to the control group(P>.05)while the menthol group value was downregulated.In 3 h,TEER of cells not treated with menthol were similar to the control group,while treatment with menthol significantly decreased TEER value(P<.05).In addition,application of menthol decreased TEER in MDCK cells earlier than in MDCK-MDR1 cells.Conclusion:Menthol but not puerarin and paeoniflorin may enhance paracellular transport and improve drug penetration of the BBB by disrupting the structure and,thereby,weakening the barrier function of TJs.
基金the National Natural Science Foundation of China(No.81874404).
文摘This study was to investigate the protective effect of paeoniflorin(PF)on hydrogen peroxide-induced injury.Firstly,“SMILES”of PF was searched in Pubchem and further was used for reverse molecular docking in Swiss Target Prediction database to obtain potential targets.Injury-related molecules were obtained from GeenCards database,and the predicted targets of PF for injury treatment were selected by Wayne diagram.For mechanism analysis,the protein-protein interactions were constructed by String,and the KEGG analysis was conducted in Webgestalt.Then,cell viability and cytotoxicity assay were established by CCK8 assay.Also,the experimental cells were allocated to control,model(200μmol·L^(−1) H_(2)O_(2)),SB20358010μmol·L^(−1)(200μmol·L^(−1) H_(2)O_(2)+SB20358010μmol·L^(−1)),PF 50μmol·L^(−1)(200μmol·L^(−1) H_(2)O_(2)+PF 50μmol·L^(−1)),and PF 100μmol·L^(−1)(200μmol·L^(−1) H_(2)O_(2)+PF 100μmol·L^(−1))groups.We measured the intracellular ROS,Hoechst 33258 staining,cell apoptosis,the levels of Bcl-xl,Bcl-2,Caspase-3,Cleaved-caspase3,Cleaved-caspase7,TRPA1,TRPV1,and the phosphorylation expression of p38MAPK.There are 96 potential targets that may be associated with PF for injury treatment.Then,we chose the“Inflammatory mediator regulation of TRP channels”pathway for the experimental verification from the first 10 KEGG pathway.In experimental verification,H_(2)O_(2) decreased the cell viability moderately(P<0.05),and 100μmol·L^(−1) PF increased the cell viability significantly(P<0.05).Depending on the difference of intracellular ROS fluorescence intensity,PF inhibited H_(2)O_(2)-induced reactive oxygen species production in Schwann cells.In Hoechst 33258 staining,PF reversed the condensed chromatin and apoptotic nuclei following H_(2)O_(2) treatment.Moreover,Flow cytometry results showed that PF could substantially inhibit H_(2)O_(2) induced apoptosis(P<0.05).Pretreatment with PF obviously reduced the levels of Caspase3,Cleaved-caspase3,Cleaved-caspase7,TRPA1,TRPV1,and the phosphorylation expression of p38MAPK after H_(2)O_(2) treatment(P<0.05),increased the levels of Bcl-2,and Bcl-xl(P<0.05).PF inhibited Schwann cell injury and apoptosis induced by hydrogen peroxide,which mechanism was linked to the inhibition of phosphorylation of p38MAPK.
基金the Natural Sciences Foun-dation of Hubei Province (No. 2007ABA135).
文摘In order to evaluate the efficacy of traditional paeonia extract paeoniflorin against optic nerve crush, 16 Brown Norway rats were divided into two groups at random, with 8 rats in each group. In paeoniaflorin-treated group, 2 mg paeoniaflorin （total volum： 1 mL） was injected into rat＇s peritoneum one time a day for a period of 8 days. Rats in untreated group were given a single dose of vehicle. The optic nerve was crushed by a special forceps for 30 s in the left eye and a sham procedure was performed in the right eye on the 2nd day after the first injection. The retrograde fluorogold labeling of ganglion cells was conducted 5 days after optic nerve crush. The whole retina was flat-mounted thereafter. The ganglion cells that survived the crush were counted under fluorescent microscope by using an automatic counting software. As compared with the contralateral eye, the survival rate of ganglion cells in the left eye increased from 40.22% to 64.53% with a significant difference found between them （t=2.55, P=0.023）. The results suggested that the paeonia extract paeoniflorin possessed a protective effect against optic nerve crush.
基金supported by the National Natural Science Foundation of China(Grant No.:82074092),Natural Science Foundation of Guangdong Province,China(Grant No.:2021A1515012219)Guangzhou University of Chinese Medicine“Double First-Class”and High-level University Discipline Collaborative Innovation Team Project,China(Grant No.:2021xk81) and Graduate Research Innovation Project of Guangzhou University of Chinese Medicine,China.
文摘Inhibiting the death receptor 3(DR3)signaling pathway in group 3 innate lymphoid cells(ILC3s)presents a promising approach for promoting mucosal repair in individuals with ulcerative colitis(UC).Paeoniflorin,a prominent component of Paeonia lactiflora Pall.,has demonstrated the ability to restore barrier function in UC mice,but the precise mechanism remains unclear.In this study,we aimed to delve into whether paeoniflorin may promote intestinal mucosal repair in chronic colitis by inhibiting DR3 signaling in ILC3s.C57BL/6 mice were subjected to random allocation into 7 distinct groups,namely the control group,the 2%dextran sodium sulfate(DSS)group,the paeoniflorin groups(25,50,and 100 mg/kg),the anti-tumor necrosis factor-like ligand 1A(anti-TL1A)antibody group,and the IgG group.We detected the expression of DR3 signaling pathway proteins and the proportion of ILC3s in the mouse colon using Western blot and flow cytometry,respectively.Meanwhile,DR3-overexpressing MNK-3 cells and 2%DSS-induced Rag1^(-/-)mice were used for verification.The results showed that paeoniflorin alleviated DSS-induced chronic colitis and repaired the intestinal mucosal barrier.Simultaneously,paeoniflorin inhibited the DR3 signaling pathway in ILC3s and regulated the content of cytokines(interleukin-17A,granulocyte-macrophage colony stimulating factor,and interleukin-22).Alternatively,paeoniflorin directly inhibited the DR3 signaling pathway in ILC3s to repair mucosal damage independently of the adaptive immune system.We additionally confirmed that paeoniflorin-conditioned medium(CM)restored the expression of tight junctions in Caco-2 cells via coculture.In conclusion,paeoniflorin ameliorates chronic colitis by enhancing the intestinal barrier in an ILC3-dependent manner,and its mechanism is associated with the inhibition of the DR3 signaling pathway.
基金the National Basic Research Priorities Program of China(No. 2005CB523402)the Natural Science Foundation of Zhejiang Province,China(No.Y204418)the Program for New Century Excellent Talents in University of China (No.NCET-06-0515)
文摘A high sensitive method based on liquid chromatography tandem mass spectrometry（LC-MS/MS） was developed and validated for the study of permeability of danshensu（DS） and paeoniflorin（PF） in Caco-2 intestinal absorption model. The DS and PF were extracted from cell culture by vacuum-lyophilizing and then separated on a Zorbax Stable Bond C18 column with 0.1% acetic acid aqueous solution and methanol as mobile phase. Detection was carried out by negative electrospray ionization（ESI ） with selected reaction monitoring（SRM） mode. The apparent permeability coefficients（Papp） of DS and PF in Caco-2 cell medium were calculated and the effects of verapamil on the coefficients Papp of the two test compounds were also illustrated. The permeability of PF was much better than that of DS when the two compounds were administrated individually. Co-administration of DS and PF led to the decrease of the transport from apical side to basolateral side for both the compounds. However, the transport in the contrary direction were accelerated. It was also observed that verapamil could accelerate the transport of the test compounds from apical side to basolateral side. However, the absorption-enhanced effect of verapamil was attenuated when DS and PF were co-administrated. These observations suggest that both passive diffusion and active efflux involved in P-gp would effect the passage of DS and PF across Caco-2 cell monolayer. At the same time, the co-administration of DS and PF to an alteration of transport behavior, which suggests that the interaction must be taken into account when ‘n-in-one＇ samples were used in Caco-2 intestinal model.
基金Supported by the National Natural Science Foundation of China(21176220,31240054)Key Technology Research and Development Project of Ningbo(2011C11023)Zhejiang Provincial Natural Science Foundation of China(Z13B060008)
文摘In order to facilitate the preparation of paeoniflorin(PF)and albiflorin(AF),two chief bioactive constituents in Paeonia lactiflora Pall(PL),induction and culture of callus from PL were studied.With a modified woody plant medium supplemented with 0.5 mg·L-16-benzylaminopurine,1.0 mg·L-1naphthylacetic acid,0.1 mg·L-1thidiazuron and 30 g·L-1sucrose,callus was induced from four kinds of explants:leaf,stems,petiole,and root.The potency to form callus varies between different explants and leaf explants exhibits the highest capacity(100%).On the other hand,root-derived callus(R-callus)produces the highest level of total amount of PF and AF,31.8 mg·g-1dry mass,which is higher than the corresponding level in the root of field cultivated PL.Furthermore,the time needed is only 40 days,remarkably shorter than the cultivation time of PL,about 4–5 years.Higher accumulation levels of PF and AF with shorter production time indicate that callus culture of PL is a promising powerful tool for production of PF and AF in the future.
基金supported by the National Natural Science Foundation of China(Project No.81773345 and 82104856)Basic and Applied Basic Research Foundation of Guangdong Province(No.SL2023A04J01298).
文摘Insulin-like growth factor-1(IGF-1)is considered as a pathogenic factor contributing to sebaceous gland dysfunction,which leads to acne vulgaris.Paeoniflorin(Pae),a bioactive monomer derived from total glycosides of paeony,has shown potential in treating various diseases.However,its anti-acne effects on human sebocytes are not well understood.In this study,we investigated the effects of Pae on acne development induced by IGF-1 in SZ95 sebocytes.Following IGF-1 stimulation,SZ95 sebocytes were exposed to Pae and then determined for proliferation,cell cycle,apoptosis,lipogenesis and pro-inflammatory cytokine secretion.We also analyzed the expression of proteins involved in the PI3K/Akt/FoxO1 and JAK2/STAT3 pathways.In vitro experiments demonstrated that Pae significantly inhibited colony formation,induced G1/S cell cycle arrest,promoted apoptosis,inhibited lipogenesis and cytokine synthesis in IGF-1-treated SZ95 sebocytes.Furthermore,Pae suppressed the phosphorylation of Akt,FoxO1,JAK2,and STAT3.Importantly,the sebo-suppressive and anti-inflammatory effects of Pae were enhanced by blocking PI3K and JAK2.In summary,our findings suggest that Pae has potent anti-proliferative and pro-apoptotic effects in SZ95 sebocytes.Additionally,Pae effectively protects against IGF-1-induced lipogenesis and inflammation by targeting the PI3K/Akt/FoxO1 and JAK2/STAT3 signaling pathways.
