[Objectives]To explore the effects of Radix Salviae Miltiorrhizae water extract and Semen Ziziphi Spinosae compound on theβ-secretase activity in mice exposed to aluminum maltolate.[Methods]A total of 60 healthy,clea...[Objectives]To explore the effects of Radix Salviae Miltiorrhizae water extract and Semen Ziziphi Spinosae compound on theβ-secretase activity in mice exposed to aluminum maltolate.[Methods]A total of 60 healthy,clean-grade SPF mice were randomly assigned to four groups based on their body weight,with each group consisting of 15 mice.The groups included a control group,a model group,treatment group 1,and treatment group 2.The control group received an equivalent dose of normal saline,while the model group and treatment groups 1 and 2 were intraperitoneally injected with 0.3 mg/kg of aluminum maltolate solution for 60 d.Additionally,treatment groups 1 and 2 were injected with 0.3 mg/kg of Radix Salviae Miltiorrhizae water extract and 0.3 mg/kg of Semen Ziziphi Spinosae compound,respectively,starting from the 31 st day for a total of 30 d.The cognitive functions of mice,specifically their learning and memory capabilities,were assessed using the Y-shaped water maze test at three distinct time points:prior to,during,and following the experimental procedure.Serum samples were collected for the analysis of various biochemical markers,including hemoglobin(Hb),total cholesterol(TC),triglycerides(TG),total protein(TP),alanine aminotransferase(ALT),and blood urea nitrogen(BUN).Additionally,brain tissues were harvested to evaluate the levels of glutathione peroxidase(GSH-PX)and acetylcholinesterase(AChE)in both serum and brain samples.The expression levels ofα-secretase,β-secretase,andγ-secretase in mouse serum were quantified using enzyme-linked immunosorbent assay(ELISA).[Results]According to the final results of the Y-shaped water maze test,the administration of therapeutic drugs to mice resulted in a gradual reduction in both the swimming time and the distance traveled to reach the platform in treatment groups 1 and 2.Additionally,the number of errors made by these treatment groups was significantly greater than that observed in the control group,with statistically significant differences(P<0.01).Among the three groups subjected to subchronic aluminum exposure,statistically significant differences were observed in the levels of Hb,TC,TG,TP,ALT,BUN,brain GSH-PX,and brain AChE(P<0.05).Furthermore,with the increasing duration of therapeutic drug administration,the levels ofβ-secretase in the brains of mice in both the treatment groups and the model group exhibited a significant decrease,while the levels ofα-secretase showed a significant increase.Additionally,the differences were statistically significant when compared to the control group(P<0.05).[Conclusions]Radix Salviae Miltiorrhizae water extract and Semen Ziziphi Spinosae compound will decrease the expression level ofβ-secretase activity in mice exposed to aluminum maltolate.展开更多
Lily-Ziziphi Spinosae Semen decoction(LZ)is known for its blood nourishing,mind calming,body sedation,and sleep promoting effects in traditional Chinese medicine.However,its material basis and underlying mechanisms ha...Lily-Ziziphi Spinosae Semen decoction(LZ)is known for its blood nourishing,mind calming,body sedation,and sleep promoting effects in traditional Chinese medicine.However,its material basis and underlying mechanisms have not yet been clearly defined.This study applies liquid chromatography-mass spectrometry,network pharmacology,and animal studies to reveal the material basis and sleep-improving mechanisms of LZ.The mixed decoction(LZ-ME)and single decoction(LZ-SE)were prepared to study their chemical components.Network pharmacology was used to predict the sleep-improving targets and signaling pathways of LZ.ICR mice were intragastrically administered saline(NC),melatonin(positive control group,0.50 mg/kg),low(12.90 g/kg),medium(25.70 g/kg),and high(38.60 g/kg)dose of LZ-ME and LZ-SH for 30 days.The results showed that LZ-ME could prolong the sleep duration and shorten the sleep latency in sodium barbiturate induced mice model.The results of chemical composition showed that total polysaccharides,total flavonoids,total saponins,and total alkaloids in LZ-ME were significantly higher than those in LZ-SE(177.20%,82.34%,30.58%,and 11.66%,respectively).A total of 58 chemical components were identified by ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-QTOF-MS),and 8 representative difference components of LZ-ME and LZ-SE were found.LZ-ME significantly increased tumor necrosis factor-α(TNF-α)in mice serum and neurotransmitterγ-aminobutyric acid(GABA)levels in mice hippocampus,decreased dopamine(DA)and glutamate(Glu)levels in mice hippocampus(P<0.05).Furthermore,LZ-ME up-regulated the abundance of the beneficial bacteria Lactobacillus and Eubacterium_R,down-regulated the abundance of the harmful bacteria Lachnospiraceae etc.The polysaccharides,flavonoids(spinosin and 6‴-feruloylspinosin)and saponins(jujuboside A and jujuboside B)were the main material bases for the sleep-promoting effects of LZ.These compounds may directly enhance levels of the GABA,reduced levels of Glu and DA and improve TNF-αlevels.And they also may indirectly regulate GABA levels by influencing the relative abundance of Lactobacillus and Eubacterium.展开更多
OBJECTIVE:To elucidate the potential mechanisms and identify bioactive components of Suanzaoren(Ziziphi Spinosae Semen)(ZSS)and it's processed products for insomnia treatment.METHODS:The principal components of ZS...OBJECTIVE:To elucidate the potential mechanisms and identify bioactive components of Suanzaoren(Ziziphi Spinosae Semen)(ZSS)and it's processed products for insomnia treatment.METHODS:The principal components of ZSS and its processed products were analyzed using UltraPerformance liquid chromatography quadrupole time-offlight mass spectrometry(UPLC-Q-TOF-MS).The effects of ZSS on insomnia were assessed in parachlorophenylalanine(PCPA)-induced rats.Additionally,to investigate the mechanisms of action in insomnia,serum and hippocampal non-targeted metabolomics were performed alongside network pharmacology and molecular docking.RESULTS:UPLC-Q-TOF-MS was used to analyze the main components of ZSS and its processed products.In the insomnia rat model induced by PCPA,both ZSS and its processed products demonstrated therapeutic effects,with the processed products showing particularly significant effects,significantly increasing the levels of the Gamma-Aminobutyric Acid and melatonin.Non-targeted metabolomics identified 23 and 10 potential differential metabolites in serum and hippocampus,respectively,involving pathways such as lipid metabolism and amino acid metabolism.Network pharmacology identified three core targets,mainly associated with pathways such as Transient Receptor Potential channel regulation,Advanced Glycation End-products-Receptor for Advanced Glycation End-products signaling pathway,and neuroactive ligand-receptor interaction.Molecular docking confirmed that the main active components of ZSS can stably bind to these core targets,thereby exerting an effect in improving insomnia.CONCLUSIONS:Combining in vivo experiments,metabolomics,and network pharmacology,we have identified potential pharmacological mechanisms by which ZSS and its derivatives mitigate insomnia.These findings provide a theoretical foundation for the further development and utilization of ZSS,supporting the innovation of new therapeutic agents for insomnia treatment.展开更多
Ziziphi Spinosae Semen(ZSS) has been used for treatment of insomnia in China for centuries. To reveal the influence of insomnia on the levels of the neurotransmitters including serotonin(5-HT), glutamic acid(Glu),γ-a...Ziziphi Spinosae Semen(ZSS) has been used for treatment of insomnia in China for centuries. To reveal the influence of insomnia on the levels of the neurotransmitters including serotonin(5-HT), glutamic acid(Glu),γ-aminobutyric acid(GABA),noradrenaline(NE) and dopamine(DA), and to study the role of ZSS aqueous extract in the treatment of insomnia, an UPLC-ESIMS/MS method was developed and validated for simultaneous determination of five neurotransmitters in the rat brain. The brain samples were pretreated by one-step direct protein precipitation with acetonitrile. The analytes were detected in positive mode with multiple reaction monitoring(MRM) and the procedure was completed in less than 10 min. The method showed a good linearity(R^2 >0.9967) with the other validation parameters were within acceptance range. The results indicated that the concentration of 5-HT, GABA and DA is significantly lower(P < 0.01) in para-chlorophenylalanine(PCPA)-induced insomnia rat model group, while Glu and NE significantly higher than those in control group(P < 0.01). Treatment with ZSS aqueous extract(4 or 8 g·kg^1·d^-1 for seven days) could ameliorate the symptoms of insomnia by significantly changing the levels of the neurotransmitter parameters mentioned above. The data obtained in this study demonstrate that ZSS aqueous extract could ameliorate the symptoms of insomnia by modulating the levels of monoamines and amino acid neurotransmitters in the brain.展开更多
Ziziphi Spinosae Semen(ZSS),a traditional Chinese medicine,is used in clinics for the treatment of insomnia in China and other Asian countries.Herein,we described for the first time a comparative pharmacokinetics stud...Ziziphi Spinosae Semen(ZSS),a traditional Chinese medicine,is used in clinics for the treatment of insomnia in China and other Asian countries.Herein,we described for the first time a comparative pharmacokinetics study of the six major compounds of ZSS in normal control(NC)and para-chlorophenylalanine(PCPA)-induced insomnia model(IM)rats that were orally administered the aqueous extract of ZSS.An ultra-high-performance liquid chromatography coupled with quadrupole orbitrap mass(UHPLC-Q-Orbitrap-MS)method was developed and validated for the simultaneous determination of coclaurine,magnoflorine,spinosin,6000-feruloylspinosin,jujuboside A(JuA),and jujuboside B(JuB)in ZSS in rat plasma.The established approach was successfully applied to a comparative pharmacokinetic study.The systemic exposures of spinosin and 6000-feruloylspinosin were decreased in the IM group compared to the NC group,while plasma clearance(CL)was significantly increased.The Tmax values of JuA and JuB in IM rats were significantly lower than those in NC rats.The T1/2 of JuA in the IM group was significantly accelerated.The pharmacokinetic parameters of coclaurine and magnoflorine were not evidently affected between the two groups.These results indicate that the pathological state of insomnia altered the plasma pharmacokinetics of spinosin,6000-feruloylspinosin,JuA,and JuB in the ZSS aqueous extract,providing an experimental basis for the role of ZSS in insomnia treatment.The comparative pharmacokinetics-based UHPLC-Q-Orbitrap-MS using full-scan mode can therefore provide a reliable and suitable means for the screening of potentially effective substances applied as quality markers of ZSS.展开更多
A sensitive,fast and comprehensive method for the quality assessment of Semen Ziziphi Spinosae(SZS)standard decoction with characterization of its chemical components was developed and validated.UPLC-Q/TOF-MS/MS syste...A sensitive,fast and comprehensive method for the quality assessment of Semen Ziziphi Spinosae(SZS)standard decoction with characterization of its chemical components was developed and validated.UPLC-Q/TOF-MS/MS system was used to identify thirty-six chemical components of SZS standard decoction which included nucleosides,phenolic acids,alkaloids,and flavonoids.Furthermore,a UPLC-PDA method was validated to simultaneously determine adenosine,protocatechuic acid,magnoflorine,catechin,protocatechin,vicenin II,spinosin,kaempferol-3-rutinoside,and 6'''-feruloylspinosin which represent four species of characteristic compounds.The qualitative method had been validated according to Chinese Pharmacopoeia(2015 edition)in terms of lineary,repeatability,recovery and stability for all analytes,with the results showing good precision,accuracy and stability.In conclusion,the method using UPLC combined with MS and PDA provided a novel way for the standardization and identification of SZS standard decoction,and also offered a basis for qualitative analysis and quality assessment of the preparations for SZS standard decoction.展开更多
Due to the scarcity of resources of Ziziphi spinosae semen （ZSS）, many inferior goods and even adulterants are generally found in medicine markets. To strengthen the quality control, HPLC fingerprint common pattern ...Due to the scarcity of resources of Ziziphi spinosae semen （ZSS）, many inferior goods and even adulterants are generally found in medicine markets. To strengthen the quality control, HPLC fingerprint common pattern established in this paper showed three main bioactive compounds in one chromatogram simultaneously. Principal component analysis based on DAD signals could discriminate adulterants and inferiorities. Principal component analysis indicated that all samples could be mainly regrouped into two main clusters according to the first principal component （PC1, redefined as Vicenin II） and the second principal component （PC2, redefined as zizyphusine）. PC1 and PC2 could explain 91.42%of the variance. Content of zizyphusine fluctuated more greatly than that of spinosin, and this result was also confirmed by the HPTLC result. Samples with low content of jujubosides and two common adulterants could not be used equivalently with authenticated ones in clinic, while one reference standard extract could substitute the crude drug in pharmaceutical production. Giving special consideration to the well-known bioactive saponins but with low response by end absorption, a fast and cheap HPTLC method for quality control of ZSS was developed and the result obtained was commensurate well with that of HPLC analysis. Samples having similar fingerprints to HPTLC common pattern targeting at saponins could be regarded as authenticated ones. This work provided a faster and cheaper way for quality control of ZSS and laid foundation for establishing a more effective quality control method for ZSS.展开更多
A simple, reproducible and reliable HPLC fingerprint analysis method of Ziziphi Spinosae Semen(ZSS) and Ziziphi Mauritianae Semen(ZMS) was established, and then it was applied to analyze samples collected from differe...A simple, reproducible and reliable HPLC fingerprint analysis method of Ziziphi Spinosae Semen(ZSS) and Ziziphi Mauritianae Semen(ZMS) was established, and then it was applied to analyze samples collected from different locations. A total of 12 common fingerprint peaks were designated in ZSS and ZMS, respectively, and five of which were definitely identified. Of these common peaks, 10 peaks were presented in both ZSS and ZMS, whereas two were characteristic for ZSS and ZMS, respectively. Furthermore, the two characteristic peaks of ZMS were definitely assigned as 6′′′-sinapoylspinosin and frangufoline by HPLC guided purification and structural elucidation. The HPLC fingerprint analysis was proved to be useful in identifying and discriminating ZSS and ZMS, which was beneficial for quality control of ZSS and its products.展开更多
Conventional quantity determination of Chinese herbal medicines by High-Performance Thin-Layer Chromatography (HPTLC) must be carried out on a TLC scanner for measuring the marker component on the HPTLC plate. General...Conventional quantity determination of Chinese herbal medicines by High-Performance Thin-Layer Chromatography (HPTLC) must be carried out on a TLC scanner for measuring the marker component on the HPTLC plate. Generally the HPTLC image of the herbal drug always takes the photograph as the permanent visual document. Considering the photo of the HPTLC image preserving all the visual quality-related parameters, there might be an alternative cost/effective approach worth studying. It would be presumably applicable to use the exclusively developed software as a virtual scanner instead of the TLC Scanner instrument to do digital scanning, integrating and calculating target components in the photo of the HPTLC image. It means digitally scanning the photo of the HPTLC image solely with software does the same thing as the instrumental scanner does. Based on this protocol, we primarily conducted the assay of the contents of the major saponins, jujuboside a and jujuboside b, in Ziziphi Spinosae Semen (Suan-Zao-Ren). Using home-developed software “digiscan” for acquiring the raw data and integrating the peaks area of the target components with “Origin Pro”, the contents of jujuboside a and jujuboside b can then be calculated respectively. The primary investigated experiment results were well compatible with official HPLC-ELSD method in Chinese Pharmacopoeia. The methodology validation and comparative analysis with HPLC confirmed that the unprecedented method is acceptable. The advantage of the recommended approach is far more simple, flexible and cost/effective than the conventional method, and it can be done at anytime and anywhere once only if the high quality HPTLC image photo is available. There would be a bright perspective for application to other herbal medicines. And the one-stop software for merging digital scanning, integration and calculation together is in progress.展开更多
Objective The objective of the study was to explore whether Suanzaoren(Semen Ziziphi Spinosae,SZS)extract could improve insomnia by inhibiting the p38 mitogenactivated protein kinase(p38MAPK)/nuclear factor-κB(NF-κB...Objective The objective of the study was to explore whether Suanzaoren(Semen Ziziphi Spinosae,SZS)extract could improve insomnia by inhibiting the p38 mitogenactivated protein kinase(p38MAPK)/nuclear factor-κB(NF-κB)signaling pathway.Methods Forty SPF-grade Sprague-Dawley(SD)rats were included in the study,with 10 randomly selected rats serving as the control group.The remaining rats were injected intraperitoneally with p-chlorophenylalanine(PCPA)for 6 days to establish an insomnia model.After successful modeling,the rats were divided into the model group,SZS extract group(3.0 g/kg),and zopiclone group(1.25 g/kg).The rats in the SZS extract and zopiclone groups were administered with the corresponding drugs via gavage for 7 days,while the rats in the control and model groups received distilled water.Sleep latency and sleep duration were recorded,and behavioral changes were observed through elevated plusmaze and open field tests.The levels of oxidative stress markers and serum inflammatory factors were measured by enzyme-linked immunosorbent assay(ELISA).The expression levels of p38 MAPK,p-p38MAPK,p-NF-κBp65,and NF-κBp65 protein in the cerebral cortex were detected by Western blot.Neuronal structures in the cerebral cortex were observed under a transmission electron microscope.Results Compared with the control group,the model group exhibited abnormal appearances,significant body mass loss(p<0.001),prolonged sleep latency and shortened sleep duration(p<0.001).The SZS extract and zopiclone groups showed significant improvements in these parameters compared with the model group.Compared with the control group,the model group showed significant reduction in total movement distance(p<0.001),fewer entries into the central zone(p<0.01),and significant decrease in rearing frequency(p<0.001);the levels of glutathione peroxidase(GSH-Px)and catalase(CAT)in the hippocampus were significantly reduced(p<0.001);the serum levels of interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),and the expression levels of p-p38MAPK and p-NF-κBp65 in the cerebral cortex were significantly increased(p<0.05).Compared with the model group,the SZS extract group showed significant increase in movement distance(p<0.01)and rearing frequency(p<0.001),significantly increased the GSH-Px and CAT levels(p<0.001),and decreased the IL-1βand TNF-αlevels(p<0.01);furthermore,the SZS extract group showed a significantly reduced p-p38MAPK and p-NF-κBp65 levels(p<0.05).The SZS extract group showed significant improvement in the neuronal structure compared with the model group.Conclusion SZS extract can inhibit the p38MAPK/NF-κB signaling pathway to improve insomnia.展开更多
Background:To facilitate the preparation of traditional Chinese medicines they are pre-mashed,i.e.,mashed in advance.However,storage conditions for pre-mashed traditional Chinese medicines are based on subjective judg...Background:To facilitate the preparation of traditional Chinese medicines they are pre-mashed,i.e.,mashed in advance.However,storage conditions for pre-mashed traditional Chinese medicines are based on subjective judgments of pharmacists,and the best storage conditions have not generally been determined.Semen Zizyphi Spinosae is a commonly used traditional Chinese medicine,and it is usually used after it is fried.The medicine needs to be mashed in a timely manner to ensure its effectiveness.The Chinese Pharmacopoeia provides a limit for its aflatoxin content.Methods:The orthogonal experimental design method optimizes the best plan for pre-mashed fried Semen Zizyphi Spinosae.Experimental conditions were defined using the L^(9)(3^(4))orthogonal design table.Four factors and three levels were used to study storage conditions.The four factors and three levels are as follows:storage temperature(10°C,20°C,and 30°C),storage humidity(45%,60%,and 75%),storage times(10,20 and 30 days),and particle sizes for the powder(coarsest,coarse and medium powders).The contents of jujuboside A,spinosin,aflatoxin B1,total aflatoxins(aflatoxins B1,B2,G1,and G2),moisture,total ash,acid value,and saponification values were measured.Results:The results demonstrated that the highest jujuboside A and spinosin contents were obtained using a storage temperature of 20°C,a storage humidity of 75%,a storage time of 10 days,and with a coarse powder particle size.Aflatoxin B1 and total aflatoxins(aflatoxins B1,B2,G1 and G2)were not detected under these conditions.Conclusion:There is no requirement for traditional Chinese medicines to be pre-mashed.This study evaluated various storage conditions for pre-mashed fried Semen Zizyphi Spinosae,and considered the influence of four factors on the contents of jujuboside A,spinolin and aflatoxin for quality control to provide a reference for other pre-mashed traditional Chinese medicines.展开更多
The integration and modularity of leaf morphological traits are fundamental to plant adaptations, yet their responses to diverse environmental pressures remain unclear. In this study, we investigate the roles of leaf ...The integration and modularity of leaf morphological traits are fundamental to plant adaptations, yet their responses to diverse environmental pressures remain unclear. In this study, we investigate the roles of leaf trait integration and modularity and how they interact with environmental factors. We analyzed geometric, traditional, and functional leaf traits across 908 individuals from 72 populations of two alpine evergreen oaks, Quercus aquifolioides Rehder & E.H. Wilson and Quercus spinosa David ex Franch., distributed throughout the Himalayan-Hengduan Mountains(HHM), employing genetic assignment as a priori. Multivariate and redundancy analyses revealed that Q. aquifolioides, which inhabits harsher environments, exhibits lower trait integration and greater morphological flexibility, allowing for dynamic adaptation to fluctuating conditions. In contrast, Q. spinosa, thriving in milder environments, demonstrates stronger integration and stability in leaf morphology, facilitating resource optimization and providing a competitive advantage. Notable differences in modularity between the two species were observed, particularly in specific leaf traits, as revealed by structural equation modeling(SEM) analysis. These results underscore the adaptive significance of leaf trait integration and modularity in extreme environments and highlight the critical role of leaf morphology in enhancing species resilience.展开更多
目的：建立刺山柑种子中没食子酸和芦丁的含量测定方法。方法采用Phnomenex色谱柱（250 mm ＆#215;4．6 mm ，5μm），以乙腈-2 mL · L -1磷酸溶液为流动相，流速为1．0 mL · min-1，检测波长为254 nm ，梯度洗脱。结果没食...目的：建立刺山柑种子中没食子酸和芦丁的含量测定方法。方法采用Phnomenex色谱柱（250 mm ＆#215;4．6 mm ，5μm），以乙腈-2 mL · L -1磷酸溶液为流动相，流速为1．0 mL · min-1，检测波长为254 nm ，梯度洗脱。结果没食子酸在1.4～14．0μg、芦丁在1.4～28．0μg范围内呈良好的线性关系，没食子酸平均回收率为99．9％，RSD为2．0％；芦丁平均回收率为100．6％， RSD为1．3％。结论 HPLC法测定没食子酸和芦丁的含量，方法简便可行，重复性和分离效果好，可为刺山柑的进一步开发利用及质量控制提供依据。展开更多
基金Supported by Undergraduate Innovation and Entrepreneurship Training Program(202410599042).
文摘[Objectives]To explore the effects of Radix Salviae Miltiorrhizae water extract and Semen Ziziphi Spinosae compound on theβ-secretase activity in mice exposed to aluminum maltolate.[Methods]A total of 60 healthy,clean-grade SPF mice were randomly assigned to four groups based on their body weight,with each group consisting of 15 mice.The groups included a control group,a model group,treatment group 1,and treatment group 2.The control group received an equivalent dose of normal saline,while the model group and treatment groups 1 and 2 were intraperitoneally injected with 0.3 mg/kg of aluminum maltolate solution for 60 d.Additionally,treatment groups 1 and 2 were injected with 0.3 mg/kg of Radix Salviae Miltiorrhizae water extract and 0.3 mg/kg of Semen Ziziphi Spinosae compound,respectively,starting from the 31 st day for a total of 30 d.The cognitive functions of mice,specifically their learning and memory capabilities,were assessed using the Y-shaped water maze test at three distinct time points:prior to,during,and following the experimental procedure.Serum samples were collected for the analysis of various biochemical markers,including hemoglobin(Hb),total cholesterol(TC),triglycerides(TG),total protein(TP),alanine aminotransferase(ALT),and blood urea nitrogen(BUN).Additionally,brain tissues were harvested to evaluate the levels of glutathione peroxidase(GSH-PX)and acetylcholinesterase(AChE)in both serum and brain samples.The expression levels ofα-secretase,β-secretase,andγ-secretase in mouse serum were quantified using enzyme-linked immunosorbent assay(ELISA).[Results]According to the final results of the Y-shaped water maze test,the administration of therapeutic drugs to mice resulted in a gradual reduction in both the swimming time and the distance traveled to reach the platform in treatment groups 1 and 2.Additionally,the number of errors made by these treatment groups was significantly greater than that observed in the control group,with statistically significant differences(P<0.01).Among the three groups subjected to subchronic aluminum exposure,statistically significant differences were observed in the levels of Hb,TC,TG,TP,ALT,BUN,brain GSH-PX,and brain AChE(P<0.05).Furthermore,with the increasing duration of therapeutic drug administration,the levels ofβ-secretase in the brains of mice in both the treatment groups and the model group exhibited a significant decrease,while the levels ofα-secretase showed a significant increase.Additionally,the differences were statistically significant when compared to the control group(P<0.05).[Conclusions]Radix Salviae Miltiorrhizae water extract and Semen Ziziphi Spinosae compound will decrease the expression level ofβ-secretase activity in mice exposed to aluminum maltolate.
基金supported by the Province Natural Science Foundation of Hunan,China(2022JJ50048)Hunan Agriculture Research System(HARS-11).
文摘Lily-Ziziphi Spinosae Semen decoction(LZ)is known for its blood nourishing,mind calming,body sedation,and sleep promoting effects in traditional Chinese medicine.However,its material basis and underlying mechanisms have not yet been clearly defined.This study applies liquid chromatography-mass spectrometry,network pharmacology,and animal studies to reveal the material basis and sleep-improving mechanisms of LZ.The mixed decoction(LZ-ME)and single decoction(LZ-SE)were prepared to study their chemical components.Network pharmacology was used to predict the sleep-improving targets and signaling pathways of LZ.ICR mice were intragastrically administered saline(NC),melatonin(positive control group,0.50 mg/kg),low(12.90 g/kg),medium(25.70 g/kg),and high(38.60 g/kg)dose of LZ-ME and LZ-SH for 30 days.The results showed that LZ-ME could prolong the sleep duration and shorten the sleep latency in sodium barbiturate induced mice model.The results of chemical composition showed that total polysaccharides,total flavonoids,total saponins,and total alkaloids in LZ-ME were significantly higher than those in LZ-SE(177.20%,82.34%,30.58%,and 11.66%,respectively).A total of 58 chemical components were identified by ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-QTOF-MS),and 8 representative difference components of LZ-ME and LZ-SE were found.LZ-ME significantly increased tumor necrosis factor-α(TNF-α)in mice serum and neurotransmitterγ-aminobutyric acid(GABA)levels in mice hippocampus,decreased dopamine(DA)and glutamate(Glu)levels in mice hippocampus(P<0.05).Furthermore,LZ-ME up-regulated the abundance of the beneficial bacteria Lactobacillus and Eubacterium_R,down-regulated the abundance of the harmful bacteria Lachnospiraceae etc.The polysaccharides,flavonoids(spinosin and 6‴-feruloylspinosin)and saponins(jujuboside A and jujuboside B)were the main material bases for the sleep-promoting effects of LZ.These compounds may directly enhance levels of the GABA,reduced levels of Glu and DA and improve TNF-αlevels.And they also may indirectly regulate GABA levels by influencing the relative abundance of Lactobacillus and Eubacterium.
文摘OBJECTIVE:To elucidate the potential mechanisms and identify bioactive components of Suanzaoren(Ziziphi Spinosae Semen)(ZSS)and it's processed products for insomnia treatment.METHODS:The principal components of ZSS and its processed products were analyzed using UltraPerformance liquid chromatography quadrupole time-offlight mass spectrometry(UPLC-Q-TOF-MS).The effects of ZSS on insomnia were assessed in parachlorophenylalanine(PCPA)-induced rats.Additionally,to investigate the mechanisms of action in insomnia,serum and hippocampal non-targeted metabolomics were performed alongside network pharmacology and molecular docking.RESULTS:UPLC-Q-TOF-MS was used to analyze the main components of ZSS and its processed products.In the insomnia rat model induced by PCPA,both ZSS and its processed products demonstrated therapeutic effects,with the processed products showing particularly significant effects,significantly increasing the levels of the Gamma-Aminobutyric Acid and melatonin.Non-targeted metabolomics identified 23 and 10 potential differential metabolites in serum and hippocampus,respectively,involving pathways such as lipid metabolism and amino acid metabolism.Network pharmacology identified three core targets,mainly associated with pathways such as Transient Receptor Potential channel regulation,Advanced Glycation End-products-Receptor for Advanced Glycation End-products signaling pathway,and neuroactive ligand-receptor interaction.Molecular docking confirmed that the main active components of ZSS can stably bind to these core targets,thereby exerting an effect in improving insomnia.CONCLUSIONS:Combining in vivo experiments,metabolomics,and network pharmacology,we have identified potential pharmacological mechanisms by which ZSS and its derivatives mitigate insomnia.These findings provide a theoretical foundation for the further development and utilization of ZSS,supporting the innovation of new therapeutic agents for insomnia treatment.
基金supported by the Natural Science Foundation of State(Nos.81603289 and 81603251)the Base Program of the Joint Training Talents of Shanxi Province(No.2017JD01)the Key Laboratory of Effective Substances Research and Utilization in TCM of Shanxi province(No.201605D111004)
文摘Ziziphi Spinosae Semen(ZSS) has been used for treatment of insomnia in China for centuries. To reveal the influence of insomnia on the levels of the neurotransmitters including serotonin(5-HT), glutamic acid(Glu),γ-aminobutyric acid(GABA),noradrenaline(NE) and dopamine(DA), and to study the role of ZSS aqueous extract in the treatment of insomnia, an UPLC-ESIMS/MS method was developed and validated for simultaneous determination of five neurotransmitters in the rat brain. The brain samples were pretreated by one-step direct protein precipitation with acetonitrile. The analytes were detected in positive mode with multiple reaction monitoring(MRM) and the procedure was completed in less than 10 min. The method showed a good linearity(R^2 >0.9967) with the other validation parameters were within acceptance range. The results indicated that the concentration of 5-HT, GABA and DA is significantly lower(P < 0.01) in para-chlorophenylalanine(PCPA)-induced insomnia rat model group, while Glu and NE significantly higher than those in control group(P < 0.01). Treatment with ZSS aqueous extract(4 or 8 g·kg^1·d^-1 for seven days) could ameliorate the symptoms of insomnia by significantly changing the levels of the neurotransmitter parameters mentioned above. The data obtained in this study demonstrate that ZSS aqueous extract could ameliorate the symptoms of insomnia by modulating the levels of monoamines and amino acid neurotransmitters in the brain.
基金This work was supported by grants from the National Natural Science Foundation of China(No:81603289,81603251)Key Laboratory of Effective Substances Research and Utilization in TCM of Shanxi province(No:201605D111004)and Key Technology Research Zhen Dong Special Project from Shanxi Science and Technology Department(No:2016ZD0105).
文摘Ziziphi Spinosae Semen(ZSS),a traditional Chinese medicine,is used in clinics for the treatment of insomnia in China and other Asian countries.Herein,we described for the first time a comparative pharmacokinetics study of the six major compounds of ZSS in normal control(NC)and para-chlorophenylalanine(PCPA)-induced insomnia model(IM)rats that were orally administered the aqueous extract of ZSS.An ultra-high-performance liquid chromatography coupled with quadrupole orbitrap mass(UHPLC-Q-Orbitrap-MS)method was developed and validated for the simultaneous determination of coclaurine,magnoflorine,spinosin,6000-feruloylspinosin,jujuboside A(JuA),and jujuboside B(JuB)in ZSS in rat plasma.The established approach was successfully applied to a comparative pharmacokinetic study.The systemic exposures of spinosin and 6000-feruloylspinosin were decreased in the IM group compared to the NC group,while plasma clearance(CL)was significantly increased.The Tmax values of JuA and JuB in IM rats were significantly lower than those in NC rats.The T1/2 of JuA in the IM group was significantly accelerated.The pharmacokinetic parameters of coclaurine and magnoflorine were not evidently affected between the two groups.These results indicate that the pathological state of insomnia altered the plasma pharmacokinetics of spinosin,6000-feruloylspinosin,JuA,and JuB in the ZSS aqueous extract,providing an experimental basis for the role of ZSS in insomnia treatment.The comparative pharmacokinetics-based UHPLC-Q-Orbitrap-MS using full-scan mode can therefore provide a reliable and suitable means for the screening of potentially effective substances applied as quality markers of ZSS.
基金financially supported by the National Key Scientific Project for China New Drug Discovery and Twelfth Five-Year Plan-preclinical study on Suanzaoren granules(No.2014ZX09304306)the Key Technologies of Common Quality Evaluation of New Drugs(Grant no.2015010201)
文摘A sensitive,fast and comprehensive method for the quality assessment of Semen Ziziphi Spinosae(SZS)standard decoction with characterization of its chemical components was developed and validated.UPLC-Q/TOF-MS/MS system was used to identify thirty-six chemical components of SZS standard decoction which included nucleosides,phenolic acids,alkaloids,and flavonoids.Furthermore,a UPLC-PDA method was validated to simultaneously determine adenosine,protocatechuic acid,magnoflorine,catechin,protocatechin,vicenin II,spinosin,kaempferol-3-rutinoside,and 6'''-feruloylspinosin which represent four species of characteristic compounds.The qualitative method had been validated according to Chinese Pharmacopoeia(2015 edition)in terms of lineary,repeatability,recovery and stability for all analytes,with the results showing good precision,accuracy and stability.In conclusion,the method using UPLC combined with MS and PDA provided a novel way for the standardization and identification of SZS standard decoction,and also offered a basis for qualitative analysis and quality assessment of the preparations for SZS standard decoction.
文摘Due to the scarcity of resources of Ziziphi spinosae semen （ZSS）, many inferior goods and even adulterants are generally found in medicine markets. To strengthen the quality control, HPLC fingerprint common pattern established in this paper showed three main bioactive compounds in one chromatogram simultaneously. Principal component analysis based on DAD signals could discriminate adulterants and inferiorities. Principal component analysis indicated that all samples could be mainly regrouped into two main clusters according to the first principal component （PC1, redefined as Vicenin II） and the second principal component （PC2, redefined as zizyphusine）. PC1 and PC2 could explain 91.42%of the variance. Content of zizyphusine fluctuated more greatly than that of spinosin, and this result was also confirmed by the HPTLC result. Samples with low content of jujubosides and two common adulterants could not be used equivalently with authenticated ones in clinic, while one reference standard extract could substitute the crude drug in pharmaceutical production. Giving special consideration to the well-known bioactive saponins but with low response by end absorption, a fast and cheap HPTLC method for quality control of ZSS was developed and the result obtained was commensurate well with that of HPLC analysis. Samples having similar fingerprints to HPTLC common pattern targeting at saponins could be regarded as authenticated ones. This work provided a faster and cheaper way for quality control of ZSS and laid foundation for establishing a more effective quality control method for ZSS.
基金National Project for Standardization of Chinese Material Medica(Grant No.ZYBZH-Y-JIN-34).
文摘A simple, reproducible and reliable HPLC fingerprint analysis method of Ziziphi Spinosae Semen(ZSS) and Ziziphi Mauritianae Semen(ZMS) was established, and then it was applied to analyze samples collected from different locations. A total of 12 common fingerprint peaks were designated in ZSS and ZMS, respectively, and five of which were definitely identified. Of these common peaks, 10 peaks were presented in both ZSS and ZMS, whereas two were characteristic for ZSS and ZMS, respectively. Furthermore, the two characteristic peaks of ZMS were definitely assigned as 6′′′-sinapoylspinosin and frangufoline by HPLC guided purification and structural elucidation. The HPLC fingerprint analysis was proved to be useful in identifying and discriminating ZSS and ZMS, which was beneficial for quality control of ZSS and its products.
文摘Conventional quantity determination of Chinese herbal medicines by High-Performance Thin-Layer Chromatography (HPTLC) must be carried out on a TLC scanner for measuring the marker component on the HPTLC plate. Generally the HPTLC image of the herbal drug always takes the photograph as the permanent visual document. Considering the photo of the HPTLC image preserving all the visual quality-related parameters, there might be an alternative cost/effective approach worth studying. It would be presumably applicable to use the exclusively developed software as a virtual scanner instead of the TLC Scanner instrument to do digital scanning, integrating and calculating target components in the photo of the HPTLC image. It means digitally scanning the photo of the HPTLC image solely with software does the same thing as the instrumental scanner does. Based on this protocol, we primarily conducted the assay of the contents of the major saponins, jujuboside a and jujuboside b, in Ziziphi Spinosae Semen (Suan-Zao-Ren). Using home-developed software “digiscan” for acquiring the raw data and integrating the peaks area of the target components with “Origin Pro”, the contents of jujuboside a and jujuboside b can then be calculated respectively. The primary investigated experiment results were well compatible with official HPLC-ELSD method in Chinese Pharmacopoeia. The methodology validation and comparative analysis with HPLC confirmed that the unprecedented method is acceptable. The advantage of the recommended approach is far more simple, flexible and cost/effective than the conventional method, and it can be done at anytime and anywhere once only if the high quality HPTLC image photo is available. There would be a bright perspective for application to other herbal medicines. And the one-stop software for merging digital scanning, integration and calculation together is in progress.
基金funded by the Research on the Mechanism of Suan Zao Ren’s(Semen Ziziphi Spinosae)Hypnotic Effect Based on TXNIP/NLRP3 Signaling Pathway(2023-ZXFZJJ-JW-071).
文摘Objective The objective of the study was to explore whether Suanzaoren(Semen Ziziphi Spinosae,SZS)extract could improve insomnia by inhibiting the p38 mitogenactivated protein kinase(p38MAPK)/nuclear factor-κB(NF-κB)signaling pathway.Methods Forty SPF-grade Sprague-Dawley(SD)rats were included in the study,with 10 randomly selected rats serving as the control group.The remaining rats were injected intraperitoneally with p-chlorophenylalanine(PCPA)for 6 days to establish an insomnia model.After successful modeling,the rats were divided into the model group,SZS extract group(3.0 g/kg),and zopiclone group(1.25 g/kg).The rats in the SZS extract and zopiclone groups were administered with the corresponding drugs via gavage for 7 days,while the rats in the control and model groups received distilled water.Sleep latency and sleep duration were recorded,and behavioral changes were observed through elevated plusmaze and open field tests.The levels of oxidative stress markers and serum inflammatory factors were measured by enzyme-linked immunosorbent assay(ELISA).The expression levels of p38 MAPK,p-p38MAPK,p-NF-κBp65,and NF-κBp65 protein in the cerebral cortex were detected by Western blot.Neuronal structures in the cerebral cortex were observed under a transmission electron microscope.Results Compared with the control group,the model group exhibited abnormal appearances,significant body mass loss(p<0.001),prolonged sleep latency and shortened sleep duration(p<0.001).The SZS extract and zopiclone groups showed significant improvements in these parameters compared with the model group.Compared with the control group,the model group showed significant reduction in total movement distance(p<0.001),fewer entries into the central zone(p<0.01),and significant decrease in rearing frequency(p<0.001);the levels of glutathione peroxidase(GSH-Px)and catalase(CAT)in the hippocampus were significantly reduced(p<0.001);the serum levels of interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),and the expression levels of p-p38MAPK and p-NF-κBp65 in the cerebral cortex were significantly increased(p<0.05).Compared with the model group,the SZS extract group showed significant increase in movement distance(p<0.01)and rearing frequency(p<0.001),significantly increased the GSH-Px and CAT levels(p<0.001),and decreased the IL-1βand TNF-αlevels(p<0.01);furthermore,the SZS extract group showed a significantly reduced p-p38MAPK and p-NF-κBp65 levels(p<0.05).The SZS extract group showed significant improvement in the neuronal structure compared with the model group.Conclusion SZS extract can inhibit the p38MAPK/NF-κB signaling pathway to improve insomnia.
基金This work was supported by the Shanghai Municipal Commission of Health and Family Planning(No.20174Y0034)the Third batch of specialized subject construction of traditional Chinese medicine in Jiading District-Traditional Chinese Medicine(No.2020-JDZYYZDXK-01).
文摘Background:To facilitate the preparation of traditional Chinese medicines they are pre-mashed,i.e.,mashed in advance.However,storage conditions for pre-mashed traditional Chinese medicines are based on subjective judgments of pharmacists,and the best storage conditions have not generally been determined.Semen Zizyphi Spinosae is a commonly used traditional Chinese medicine,and it is usually used after it is fried.The medicine needs to be mashed in a timely manner to ensure its effectiveness.The Chinese Pharmacopoeia provides a limit for its aflatoxin content.Methods:The orthogonal experimental design method optimizes the best plan for pre-mashed fried Semen Zizyphi Spinosae.Experimental conditions were defined using the L^(9)(3^(4))orthogonal design table.Four factors and three levels were used to study storage conditions.The four factors and three levels are as follows:storage temperature(10°C,20°C,and 30°C),storage humidity(45%,60%,and 75%),storage times(10,20 and 30 days),and particle sizes for the powder(coarsest,coarse and medium powders).The contents of jujuboside A,spinosin,aflatoxin B1,total aflatoxins(aflatoxins B1,B2,G1,and G2),moisture,total ash,acid value,and saponification values were measured.Results:The results demonstrated that the highest jujuboside A and spinosin contents were obtained using a storage temperature of 20°C,a storage humidity of 75%,a storage time of 10 days,and with a coarse powder particle size.Aflatoxin B1 and total aflatoxins(aflatoxins B1,B2,G1 and G2)were not detected under these conditions.Conclusion:There is no requirement for traditional Chinese medicines to be pre-mashed.This study evaluated various storage conditions for pre-mashed fried Semen Zizyphi Spinosae,and considered the influence of four factors on the contents of jujuboside A,spinolin and aflatoxin for quality control to provide a reference for other pre-mashed traditional Chinese medicines.
基金supported by the National Science Foundation of China(No.42071060)to Fang Du.
文摘The integration and modularity of leaf morphological traits are fundamental to plant adaptations, yet their responses to diverse environmental pressures remain unclear. In this study, we investigate the roles of leaf trait integration and modularity and how they interact with environmental factors. We analyzed geometric, traditional, and functional leaf traits across 908 individuals from 72 populations of two alpine evergreen oaks, Quercus aquifolioides Rehder & E.H. Wilson and Quercus spinosa David ex Franch., distributed throughout the Himalayan-Hengduan Mountains(HHM), employing genetic assignment as a priori. Multivariate and redundancy analyses revealed that Q. aquifolioides, which inhabits harsher environments, exhibits lower trait integration and greater morphological flexibility, allowing for dynamic adaptation to fluctuating conditions. In contrast, Q. spinosa, thriving in milder environments, demonstrates stronger integration and stability in leaf morphology, facilitating resource optimization and providing a competitive advantage. Notable differences in modularity between the two species were observed, particularly in specific leaf traits, as revealed by structural equation modeling(SEM) analysis. These results underscore the adaptive significance of leaf trait integration and modularity in extreme environments and highlight the critical role of leaf morphology in enhancing species resilience.
文摘目的：建立刺山柑种子中没食子酸和芦丁的含量测定方法。方法采用Phnomenex色谱柱（250 mm ＆#215;4．6 mm ，5μm），以乙腈-2 mL · L -1磷酸溶液为流动相，流速为1．0 mL · min-1，检测波长为254 nm ，梯度洗脱。结果没食子酸在1.4～14．0μg、芦丁在1.4～28．0μg范围内呈良好的线性关系，没食子酸平均回收率为99．9％，RSD为2．0％；芦丁平均回收率为100．6％， RSD为1．3％。结论 HPLC法测定没食子酸和芦丁的含量，方法简便可行，重复性和分离效果好，可为刺山柑的进一步开发利用及质量控制提供依据。
