The alleviation of chemotherapy-induced myelosuppression is an integral part of sustained and effective cancer therapy.Although the role of the hematopoietic microenvironment in the regulation of hematopoietic stem/pr...The alleviation of chemotherapy-induced myelosuppression is an integral part of sustained and effective cancer therapy.Although the role of the hematopoietic microenvironment in the regulation of hematopoietic stem/progenitor cells(HSPCs)has been widely studied,no drugs that improve hematopoiesis by targeting and modulating the hematopoietic microenvironment have been used clinically.Here,we show that the active small molecule icaritin(ICT)from the Chinese herb Epimedium brevicornum Maxim effectively alleviates chemotherapy-induced hemocytopenia in both mouse and zebrafish models.We demonstrated that ICT enhanced the number and hematopoietic function of HSPCs and that the beneficial effects of ICT occurred indirectly.Single-cell sequencing analysis confirmed that the target cells of ICT in the bone marrow microenvironment were mesenchymal stromal cells(MSCs).In addition,peroxiredoxin 1(PRDX1)was identified as a direct target of ICT.Furthermore,ICT stimulated MSCs to express the effector molecule C-X-C motif chemokine ligand 12(CXCL12)through the PRDX1-reactive oxygen species(ROS)-mitogen-activated protein kinase(MAPK)signaling axis,thereby increasing the number and function of HSPCs.These results suggest that ICT is a promising compound for achieving targeted modulation of the hematopoietic microenvironment to restore hematopoiesis after chemotherapy.展开更多
Icaritin, a prenylflavonoid derivative from Epimedium Genus, has been shown to exhibit many pharmacological and biological activities. However, the function and the underlying mechanisms of icaritin in human non-small...Icaritin, a prenylflavonoid derivative from Epimedium Genus, has been shown to exhibit many pharmacological and biological activities. However, the function and the underlying mechanisms of icaritin in human non-small cell lung cancer have not been fully elucidated. The purpose of this study was to investigate the anticancer effects of icaritin on A549 cells and explore the underlying molecular mechanism. The cell viability after icaritin treatment was tested by MTT assay. The cell cycle distribution, apoptosis and reactive oxygen species（ROS） levels were analyzed by flow cytometry. The mRNA and protein expression levels of the genes involved in proliferation and apoptosis were respectively detected by RT-PCR and Western blotting. The results demonstrated that icaritin induced cell cycle arrest at S phase, and down-regulated the expression levels of S regulatory proteins such as Cyclin A and CDK2. Icaritin also induced cell apoptosis characterized by positive Hoechst 33258 staining, accumulation of the Annexin V-positive cells, increased ROS level and alteration in Bcl-2 family proteins expression. Moreover, icaritin induced sustained phosphorylation of ERK and p38 MAPK. These findings suggested that icaritin might be a new potent inhibitor by inducing S phase arrest and apoptosis in human lung carcinoma A549 cells.展开更多
Multidrug resistance(MDR) is one of the major obstacles in cancer chemotherapy. Our previous study has shown that icariin could reverse MDR in MG-63 doxorubicin-resistant(MG-63/DOX) cells. It is reported that icariin ...Multidrug resistance(MDR) is one of the major obstacles in cancer chemotherapy. Our previous study has shown that icariin could reverse MDR in MG-63 doxorubicin-resistant(MG-63/DOX) cells. It is reported that icariin is usually metabolized to icariside II and icaritin. Herein, we investigated the effects of icariin, icariside Ⅱ, and icaritin(ICT) on reversing MDR in MG-63/DOX cells. Among these compounds, ICT exhibited strongest effect and showed no obvious cytotoxicity effect on both MG-63 and MG-63/DOX cells ranging from 1 to 10 μmol·L^(-1). Furthermore, ICT increased accumulation of rhodamine 123 and 6-carboxyfluorescein diacetate and enhanced DOX-induced apoptosis in MG-63/DOX cells in a dose-dependent manner. Further studies demonstrated that ICT decreased the m RNA and protein levels of multidrug resistance protein 1(MDR1) and multidrug resistance-associated protein 1(MRP1). We also verified that blockade of STAT3 phosphorylation was involved in the reversal effect of multidrug resistance in MG-63/DOX cells. Taken together, these results indicated that ICT may be a potential candidate in chemotherapy for osteosarcoma.展开更多
Traditional treatments for advanced hepatocellular carcinoma(HCC),such as surgical resection,transplantation,radiofrequency ablation,and chemotherapy are unsatisfactory,and therefore the exploration of powerful therap...Traditional treatments for advanced hepatocellular carcinoma(HCC),such as surgical resection,transplantation,radiofrequency ablation,and chemotherapy are unsatisfactory,and therefore the exploration of powerful therapeutic strategies is urgently needed.Immunotherapy has emerged as a promising strategy for advanced HCC treatment due to its minimal side effects and long-lasting therapeutic memory effects.Recent studies have demonstrated that icaritin could serve as an immunomodulator for effective immunotherapy of advanced HCC.Encouragingly,in 2022,icaritin soft capsules were approved by the National Medical Products Administration(NMPA)of China for the immunotherapy of advanced HCC.However,the therapeutic efficacy of icaritin in clinical practice is impaired by its poor bioavailability and unfavorable in vivo delivery efficiency.Recently,functionalized drug delivery systems including stimuli-responsive nanocarriers,cell membrane-coated nanocarriers,and living cell-nanocarrier systems have been designed to overcome the shortcomings of drugs,including the low bioavailability and limited delivery efficiency as well as side effects.Taken together,the development of icaritin-based nanomedicines is expected to further improve the immunotherapy of advanced HCC.Herein,we compared the different preparation methods for icaritin,interpreted the HCC immune microenvironment and the mechanisms underlying icaritin for treatment of advanced HCC,and discussed both the design of icaritin-based nanomedicines with high icaritin loading and the latest progress in icaritinbased nanomedicines for advanced HCC immunotherapy.Finally,the prospects to promote further clinical translation of icaritin-based nanomedicines for the immunotherapy of advanced HCC were proposed.展开更多
OBJECTIVE Skeletal muscle undergoes rapid and profound atrophy in response to decreased mechanical loading,e.g.,limb immobilization and bed rest.Phosphatidylinositol 3 kinase(PI3K)/Akt signaling pathway is critical fo...OBJECTIVE Skeletal muscle undergoes rapid and profound atrophy in response to decreased mechanical loading,e.g.,limb immobilization and bed rest.Phosphatidylinositol 3 kinase(PI3K)/Akt signaling pathway is critical for regulating the balance between protein synthesis and degradation during disuse/inactivity-induced muscle atrophy.The present study aimed to investigate whether natural product Icaritin(ICT)required PI3K/Akt signaling to exert counteractive effect on skeletal muscle atrophy following mechanical unloading.METHODS Two oral dosages of ICT(80and 120mg·kg-1·d-1)were administrated daily to adult male rats with or without daily injection of PI3K/Akt signaling inhibitor wortmannin(15μg·kg-1·d-1)during 28-d hindlimb suspension(HS).Ex vivo muscle functional testing,histological and immunohistochemical analysis were performed to determine the changes of soleus muscle function,mean muscle fiber cross-sectional area(CSA)and fiber type distribution.Western blot and real-time PCR analysis were also performed to evaluate the protein or mRNA expression of the markers involved in PI3K/Akt signaling pathway.RESULTS After 28-d HS,soleus muscle underwent profound muscle atrophy(-52.7% muscle mass vs.pre-HS baseline).The high dose ICT treatment significantly attenuated the decreases in soleus muscle mass(+22.6% vs.HS),muscle fiber CSA(+52.8% vs.HS),as well as the muscle functional testing parameters during the unloading.Molecularly,the high dose ICT treatment significantly attenuated the decreases in protein synthesis markers at protein levels(phosphorylation of Akt and its downstream proteins)during the unloading,whereas the increases in protein degradation markers at mRNA(atrogin-1and MuRF-1)and protein(nuclear FOXO1 and FOXO3a)levels during the unloading were significantly attenuated by the high dose ICT treatment.The low dose ICT treatment moderately attenuated the above changes induced by the unloading.Mechanistically,Wortmannin could abolish the above effects of ICT.CONCLUSION ICT requires participation of PI3K/Akt signaling to counteract skeletal muscle atrophy following mechanical unloading in a dose-dependent manner.展开更多
Sorafenib remains the standard systemic treatment for advanced human hepatocellular carcinoma(HCC).However,the low response rate,high recurrence,and high progression limit the therapeutic efficacy.Therefore,a combinat...Sorafenib remains the standard systemic treatment for advanced human hepatocellular carcinoma(HCC).However,the low response rate,high recurrence,and high progression limit the therapeutic efficacy.Therefore,a combination therapy strategy was advanced to strengthen the antitumor effects of sorafenib.In the present study,we aimed to evaluate whether icaritin could enhance the inhibitory effects of sorafenib on HCC cells and clarify the underlying mechanism.The cell viability was evaluated via MTT assay,and the synergistic inhibitory effects of sorafenib and icaritin were verified by calculating the combination index(CI).Their combined effects on cell proliferation or apoptosis were investigated using colony formation assay and flow cytometry.Mitochondrial membrane potential(MMP)was detected by flow cytometric assay.The protein expressions associated with the apoptotic pathway were determined by Western blotting analysis.The data demonstrated that sorafenib and icaritin exerted synergistic inhibitory effects on cell viability(CI<1).Icaritin enhanced the inhibitory effect of sorafenib on colony formation and sorafenib-induced apoptosis of HCC cells.We discovered a reduced level of antiapoptotic Bcl-2 and an elevated level of proapoptotic protein Bax when the cells were exposed to the combination.The effect of cleaved and activated PARP was also enhanced.Cleaved caspase-9 and cleaved caspase-3 were increased markedly in the combination group.Furthermore,the combination of icaritin and sorafenib significantly increased the loss of MMP compared with the single treatment group and induced the release of cytochrome c from the mitochondria to the cytosol.These findings indicated that icaritin could enhance sorafenib-induced cytotoxicity and trigger sorafenib-induced apoptosis through a mitochondria-dependent pathway.展开更多
The authors would like to apologize that,due to their neglect,some mistakes in the"2.Materials and methods"section need to be corrected.(1)The unit of DMAPP and flavonoid substrates in the“2.3.Enzymatic ass...The authors would like to apologize that,due to their neglect,some mistakes in the"2.Materials and methods"section need to be corrected.(1)The unit of DMAPP and flavonoid substrates in the“2.3.Enzymatic assays of prenyltransferases"section was not correctly written.The original description“200 mmol/L dimethylallyl pyrophosphate(DMAPP),200 mmol/L naringenin,kaempferol,or kaempferide as substrate"should be corrected to“200μmol/L dimethylallyl pyrophosphate(DMAPP),200μmol/L naringenin,kaempferol,or kaempferide as substrate"(2)The concentration of acetate acid in“2.8.Chemical analysis"was not correctly written.The original description“0.01%acetate acid(A)"should be corrected to“0.1%acetate acid(A)".The authors declare that these corrections do not change the results or conclusions of this paper.The authors sincerely apologize for this omission.展开更多
Background and Aims:Lipid accumulation is the major characteristic of non-alcoholic fatty liver disease,the prevalence of which continues to rise.We aimed to investigate the effects and mechanisms of icaritin on lipid...Background and Aims:Lipid accumulation is the major characteristic of non-alcoholic fatty liver disease,the prevalence of which continues to rise.We aimed to investigate the effects and mechanisms of icaritin on lipid accumulation.Methods:Cells were treated with icaritin at 0.7,2.2,6.7,or 20μM for 24 h.The effects on lipid accumulation in L02 and Huh-7 cells were detected by Bodipy and oil red O staining,respectively.Mitochondria biogenesis of L02 cells was detected by MitoTracker Orange staining.Glucose uptake and adenosine triphosphate content of 3T3-L1 adipocytes and C2C12 myotubes were detected.The expression levels of proteins in the adenosine 5′-monophosphate-activated protein kinase(AMPK)signaling pathway,biomarkers of autophagy,and mitochondria biogenesis were measured by western blotting.LC3 puncta were detected by immunofluorescence.Results:Icaritin significantly attenuated lipid accumulation in L02 and Huh-7 cells and boosted the mitochondria biogenesis of L02 cells.Icaritin enhanced glucose uptake,decreased adenosine triphosphate content,and activated the AMPK signaling pathway in 3T3-L1 adipocytes and C2C12 myotubes.Icaritin boosted autophagy and also enhanced the initiation of autophagic flux in 3T3-L1 preadipocytes and C2C12 myoblasts.However,icaritin decreased autophagy and promoted mitochondria biogenesis in 3T3-L1 adipocytes and C2C12 myotubes.Conclusions:Icaritin attenuates lipid accumulation by increasing energy expenditure and regulating autophagy by activating the AMPK pathway.展开更多
This study focuses on the preparation and enzymic hydrolysis of an icariin/bcyclodextrin inclusion complex to efficiently generate icaritin.The physical characteristics of the inclusion complex were evaluated by diffe...This study focuses on the preparation and enzymic hydrolysis of an icariin/bcyclodextrin inclusion complex to efficiently generate icaritin.The physical characteristics of the inclusion complex were evaluated by differential scanning calorimetry(DSC).Enzymatic hydrolysis was optimized for the conversion of icariin/b-cyclodextrin complex to icaritin by Box–Behnken statistical design.The inclusion complex formulation increased the solubility of icariin approximately 17-fold,from 29.2 to 513.5 mg/mL at 60℃.The optimum conditions were predicted by Box–Behnken statistical design as follows:60℃,pH 7.0,the ratio of enzyme/substrate(1:1.1)and reaction time 7 h.Under the optimal conditions the conversion of icariin was 97.91%and the reaction time was decreased by 68%compared with that without b-CD inclusion.Product analysis by melting point,ESI-MS,UV,IR,1H NMR and 13C NMR confirmed the authenticity of icaritin with a purity of 99.3%and a yield of 473 mg of icaritin from 1.1 g icariin.展开更多
Objective： To investigate the protective effects of icaritin （ICT）, one of the active ingredients in Epimedii Folium, on mouse model of cerebral ischemia-reperfusion （I/R） in vivo. Methods： ICR mice were subject...Objective： To investigate the protective effects of icaritin （ICT）, one of the active ingredients in Epimedii Folium, on mouse model of cerebral ischemia-reperfusion （I/R） in vivo. Methods： ICR mice were subjected to an I h transient middle cerebral artery occlusion （MCAO） and fol- lowed by 24 h of reperfusion. Neurological deficits, infarct volume, brain edema and survive rate were measured, respectively. The levels of brain IL-1β, TNF-a, ROS and DNA-binding activity of NF-KB p65 were measured by ELISA kits. The levels of malondialdehyde （MDA） and activities of superoxide dismu- tase （SOD） were detected by spectrophotometry, and the release of nitric oxide （NO） were detected by Griess kit. Results： ICT markedly reduced the neurological deficit scores, brain edema, infarct volume and increased the survival rate of the cerebral I/R mice. The expression of IL-Iβ, TNF-α, NO, MDA and DNA-binding activity of NF-KB p65 were significantly inhibited by ICT, while the activity of SOD were up-regulated at the same time. Conclusion： ICT possessed significant neuroprotective effects in cerebral I/R mice, which might be related to prevent neuroinflammatory and oxidative damage.展开更多
Objective Icaritin is the main aglycone and also active intestinal metabolite of prenylflavonoids from the Chinese materia medica Epimedii Herba. Modern pharmacological studies have demonstrated that icaritin has a wi...Objective Icaritin is the main aglycone and also active intestinal metabolite of prenylflavonoids from the Chinese materia medica Epimedii Herba. Modern pharmacological studies have demonstrated that icaritin has a wide range of biological activities. However, its metabolites and biotransformation pathways have not yet been comprehensively investigated. The present study aims to identify icaritin metabolites in rats by using a sensitive and effective LC-MS/MS method. Methods The plasma and urine samples of rats were collected before （blank） and after oral administration of icaritin, and subjected to liquid-liquid extraction with ethyl acetate. The full-scan LC-MS chromatograms of the plasma and urine samples were compared with those of blank samples to detect the possible metabolites, which were later detected by their product ion spectra. Results A total of 23 metabolites were identified, and conjugated icaritins produced by glucuronidation, glycosylation, and sulfation were its major metabolites. Minor demethylation, hydrogenation, and oxidation metabolites were also found. Conclusion Phase II metabolism is the main metabolic pathway of icaritin.展开更多
Tumor relapse and metastasis are the major causes of mortality associated with urothelial cancer.In the tumor microenvironment,negative regulatory molecules and various immune cell subtypes suppress antitumor immunity...Tumor relapse and metastasis are the major causes of mortality associated with urothelial cancer.In the tumor microenvironment,negative regulatory molecules and various immune cell subtypes suppress antitumor immunity.The inflammatory microenvironment,associated with neutrophils and neutrophil extracellular traps(NETs),promotes tumor metastasis.However,no drugs are currently available to specifically inhibit neutrophils and NETs.In this study,we first demonstrated that icaritin(ICT),a Chinese herbal remedy that is a first-line treatment for advanced and incurable hepatocellular carcinoma,reduces NETs caused by suicidal NETosis and prevents neutrophil infiltration in the tumor microenvironment.Mechanistically,ICT binds to and inhibits the expression of PADI2 in neutrophils,thereby suppressing PADI2-mediated histone citrullination.Moreover,ICT inhibits ROS generation,suppresses the MAPK signaling pathway,and inhibits NET-induced tumor metastasis.Simultaneously,ICT inhibits tumoral PADI2-mediated histone citrullination,which consequently suppresses the transcription of neutrophil-recruiting genes such as GM-CSF and IL-6.The downregulation of IL-6 expression,in turn,forms a regulatory feedback loop through the JAK2/STAT3/IL-6 axis.Through a retrospective study of clinical samples,we found a correlation between neutrophils,NETs,UCa prognosis,and immune evasion.Combining ICT with immune checkpoint inhibitors may have synergistic effects.In summary,our study demonstrated that ICT could be a novel inhibitor of NETs and a novel UCa treatment.展开更多
Objective: To identify the in vivo metabolites of icaritin and speculate its metabolic profiling in rats.Methods: The plasma, bile, urine, and feces of rats were collected after orally administration of icaritin at a ...Objective: To identify the in vivo metabolites of icaritin and speculate its metabolic profiling in rats.Methods: The plasma, bile, urine, and feces of rats were collected after orally administration of icaritin at a dose of 100 mg/kg and detected by an ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC/Q-TOF-MS/MS) in both positive and negative modes.The data of treated and control groups were compared and analyzed with the aid of Metabolynx XS software.Results: A total of 25 metabolites were identified in the biosamples, and 14 of them were reported for the first time to our knowledge.Conclusion: The main metabolite types of icaritin in rats were glucuronide conjugation, methylation, hydroxylation, reduction, and acetylation.展开更多
Bone tissue engineering provides a promising strategy for the treatment of bone defects.Nonetheless,the clinical utilization of biomaterial-based scaffolds is constrained by their inadequate mechanical strength and ab...Bone tissue engineering provides a promising strategy for the treatment of bone defects.Nonetheless,the clinical utilization of biomaterial-based scaffolds is constrained by their inadequate mechanical strength and absence of osteo-inductive properties.Here,we proposed to endow nano-scaffold(NS)constructed by coaxial electrospinning technique with enhanced osteogenic bioactivities and mechanical properties by incorporating biocompatible magnetic iron oxide nanoparticles(IONPs)and icaritin(ICA).Four types of nano-scaffolds(NS,ICA@NS,NS-IONPs and ICA@NS-IONPs)were prepared.The incorporation of ICA and IONPs minimally impact their surface morphological and chemical properties.IONPs enhanced the mechanical properties of NS scaffolds,including hardness,tensile strength,and elastic modulus.In vitro assessments demonstrated that ICA@NS-IONPs exhibited enhanced osteogenic bioactivities towards mouse calvarial pre-osteoblast cell line MC3T3-E1 as evidenced by detecting the alkaline phosphatase(ALP)activity level,expressions of osteogenesis-related genes and proteins as well as mineralized nodule formation.Mechanistic investigations revealed that MEK/ERK(MAP kinase-ERK kinase(MEK)/extracellularsignal-regulated kinase(ERK))signaling pathway could offer a plausible explanation for the osteogenic differentiation of MC3T3-E1 cells induced by ICA@NS-IONPs.Furthermore,the implantation of nano-scaffolds in rat skull defects exhibited a substantial improvement in in vivo bone regeneration.Therefore,IONPs and ICA incorporated coaxial electrospinning nano-scaffolds present a novel strategy for the optimization of scaffolds for bone tissue engineering.展开更多
中枢神经系统疾病(central nervous system diseases,CNSD)已成为日益严峻的全球性健康难题。中医药在CNSD方面展现独特的疗效与丰富的临床实践积累。淫羊藿Epimedii Folium作为一味传统中药,通过多靶点调控、多通路干预及多途径作用机...中枢神经系统疾病(central nervous system diseases,CNSD)已成为日益严峻的全球性健康难题。中医药在CNSD方面展现独特的疗效与丰富的临床实践积累。淫羊藿Epimedii Folium作为一味传统中药,通过多靶点调控、多通路干预及多途径作用机制在神经系统疾病的治疗中发挥重要作用。随着淫羊藿药理作用的不断深入研究,发现其活性成分淫羊藿苷、淫羊藿素、淫羊藿次苷Ⅱ等是治疗CNSD的主要物质基础。淫羊藿活性成分可通过抑制炎症反应与氧化应激、抑制β-淀粉样蛋白沉积与微管相关蛋白异常磷酸化、抑制细胞凋亡、调节细胞自噬、改善神经元功能、调节神经递质等途径在阿尔茨海默病、帕金森病、癫痫、多发性硬化、脑缺血再灌注损伤、焦虑障碍、抑郁症、精神分裂症等CNSD中发挥作用。通过总结淫羊藿活性成分及其复方干预CNSD的作用机制,为进一步开发药物与临床运用奠定理论基础,为淫羊藿在CNSD研究中提供新的治疗方案和思路。展开更多
基金funded by the National Natural Science Foundation of China(82141203,82374086,and 82274172)the Shanghai Municipal Science and Technology Major Project(ZD2021CY001)+2 种基金the Three-year Action Plan for Shanghai TCM Development and Inheritance Program(ZY(2021-2023)-0401)the Innovation Team and Talents Cultivation Program of the National Administration of Traditional Chinese Medicine(ZYYCXTDD-202004)the Organizational Key Research and Development Program of Shanghai University of Traditional Chinese Medicine(2023YZZ02).
文摘The alleviation of chemotherapy-induced myelosuppression is an integral part of sustained and effective cancer therapy.Although the role of the hematopoietic microenvironment in the regulation of hematopoietic stem/progenitor cells(HSPCs)has been widely studied,no drugs that improve hematopoiesis by targeting and modulating the hematopoietic microenvironment have been used clinically.Here,we show that the active small molecule icaritin(ICT)from the Chinese herb Epimedium brevicornum Maxim effectively alleviates chemotherapy-induced hemocytopenia in both mouse and zebrafish models.We demonstrated that ICT enhanced the number and hematopoietic function of HSPCs and that the beneficial effects of ICT occurred indirectly.Single-cell sequencing analysis confirmed that the target cells of ICT in the bone marrow microenvironment were mesenchymal stromal cells(MSCs).In addition,peroxiredoxin 1(PRDX1)was identified as a direct target of ICT.Furthermore,ICT stimulated MSCs to express the effector molecule C-X-C motif chemokine ligand 12(CXCL12)through the PRDX1-reactive oxygen species(ROS)-mitogen-activated protein kinase(MAPK)signaling axis,thereby increasing the number and function of HSPCs.These results suggest that ICT is a promising compound for achieving targeted modulation of the hematopoietic microenvironment to restore hematopoiesis after chemotherapy.
基金supported by grants from Wuhan Municipal Science and Technology Research Project,China(No.201260523185)the Public Science and Technology Research Funds Projects of Ocean,China(No.201005013)
文摘Icaritin, a prenylflavonoid derivative from Epimedium Genus, has been shown to exhibit many pharmacological and biological activities. However, the function and the underlying mechanisms of icaritin in human non-small cell lung cancer have not been fully elucidated. The purpose of this study was to investigate the anticancer effects of icaritin on A549 cells and explore the underlying molecular mechanism. The cell viability after icaritin treatment was tested by MTT assay. The cell cycle distribution, apoptosis and reactive oxygen species（ROS） levels were analyzed by flow cytometry. The mRNA and protein expression levels of the genes involved in proliferation and apoptosis were respectively detected by RT-PCR and Western blotting. The results demonstrated that icaritin induced cell cycle arrest at S phase, and down-regulated the expression levels of S regulatory proteins such as Cyclin A and CDK2. Icaritin also induced cell apoptosis characterized by positive Hoechst 33258 staining, accumulation of the Annexin V-positive cells, increased ROS level and alteration in Bcl-2 family proteins expression. Moreover, icaritin induced sustained phosphorylation of ERK and p38 MAPK. These findings suggested that icaritin might be a new potent inhibitor by inducing S phase arrest and apoptosis in human lung carcinoma A549 cells.
基金supported by the National Natural Science Foundation of China(Nos.81673554 and 81503211)Natural Science Foundation of Jiangsu Province(No.BK20160763)+1 种基金the Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD)the Program for Changjiang Scholars and Innovative Research Team in University(No.IRT_15R63)
文摘Multidrug resistance(MDR) is one of the major obstacles in cancer chemotherapy. Our previous study has shown that icariin could reverse MDR in MG-63 doxorubicin-resistant(MG-63/DOX) cells. It is reported that icariin is usually metabolized to icariside II and icaritin. Herein, we investigated the effects of icariin, icariside Ⅱ, and icaritin(ICT) on reversing MDR in MG-63/DOX cells. Among these compounds, ICT exhibited strongest effect and showed no obvious cytotoxicity effect on both MG-63 and MG-63/DOX cells ranging from 1 to 10 μmol·L^(-1). Furthermore, ICT increased accumulation of rhodamine 123 and 6-carboxyfluorescein diacetate and enhanced DOX-induced apoptosis in MG-63/DOX cells in a dose-dependent manner. Further studies demonstrated that ICT decreased the m RNA and protein levels of multidrug resistance protein 1(MDR1) and multidrug resistance-associated protein 1(MRP1). We also verified that blockade of STAT3 phosphorylation was involved in the reversal effect of multidrug resistance in MG-63/DOX cells. Taken together, these results indicated that ICT may be a potential candidate in chemotherapy for osteosarcoma.
基金supported by the National Natural Science Foundation of China(52103181,81873196)the Sino-German Center for Research Promotion(GZ1505)+1 种基金the Fundamental Research Funds for the Central Universities(22120220075)the China Scholarship Council(201908320330)。
文摘Traditional treatments for advanced hepatocellular carcinoma(HCC),such as surgical resection,transplantation,radiofrequency ablation,and chemotherapy are unsatisfactory,and therefore the exploration of powerful therapeutic strategies is urgently needed.Immunotherapy has emerged as a promising strategy for advanced HCC treatment due to its minimal side effects and long-lasting therapeutic memory effects.Recent studies have demonstrated that icaritin could serve as an immunomodulator for effective immunotherapy of advanced HCC.Encouragingly,in 2022,icaritin soft capsules were approved by the National Medical Products Administration(NMPA)of China for the immunotherapy of advanced HCC.However,the therapeutic efficacy of icaritin in clinical practice is impaired by its poor bioavailability and unfavorable in vivo delivery efficiency.Recently,functionalized drug delivery systems including stimuli-responsive nanocarriers,cell membrane-coated nanocarriers,and living cell-nanocarrier systems have been designed to overcome the shortcomings of drugs,including the low bioavailability and limited delivery efficiency as well as side effects.Taken together,the development of icaritin-based nanomedicines is expected to further improve the immunotherapy of advanced HCC.Herein,we compared the different preparation methods for icaritin,interpreted the HCC immune microenvironment and the mechanisms underlying icaritin for treatment of advanced HCC,and discussed both the design of icaritin-based nanomedicines with high icaritin loading and the latest progress in icaritinbased nanomedicines for advanced HCC immunotherapy.Finally,the prospects to promote further clinical translation of icaritin-based nanomedicines for the immunotherapy of advanced HCC were proposed.
基金The project supported by National Natural Science Foundation of China(81201406)Direct Grant for Research,The Chinese University of Hong Kong(4054138)
文摘OBJECTIVE Skeletal muscle undergoes rapid and profound atrophy in response to decreased mechanical loading,e.g.,limb immobilization and bed rest.Phosphatidylinositol 3 kinase(PI3K)/Akt signaling pathway is critical for regulating the balance between protein synthesis and degradation during disuse/inactivity-induced muscle atrophy.The present study aimed to investigate whether natural product Icaritin(ICT)required PI3K/Akt signaling to exert counteractive effect on skeletal muscle atrophy following mechanical unloading.METHODS Two oral dosages of ICT(80and 120mg·kg-1·d-1)were administrated daily to adult male rats with or without daily injection of PI3K/Akt signaling inhibitor wortmannin(15μg·kg-1·d-1)during 28-d hindlimb suspension(HS).Ex vivo muscle functional testing,histological and immunohistochemical analysis were performed to determine the changes of soleus muscle function,mean muscle fiber cross-sectional area(CSA)and fiber type distribution.Western blot and real-time PCR analysis were also performed to evaluate the protein or mRNA expression of the markers involved in PI3K/Akt signaling pathway.RESULTS After 28-d HS,soleus muscle underwent profound muscle atrophy(-52.7% muscle mass vs.pre-HS baseline).The high dose ICT treatment significantly attenuated the decreases in soleus muscle mass(+22.6% vs.HS),muscle fiber CSA(+52.8% vs.HS),as well as the muscle functional testing parameters during the unloading.Molecularly,the high dose ICT treatment significantly attenuated the decreases in protein synthesis markers at protein levels(phosphorylation of Akt and its downstream proteins)during the unloading,whereas the increases in protein degradation markers at mRNA(atrogin-1and MuRF-1)and protein(nuclear FOXO1 and FOXO3a)levels during the unloading were significantly attenuated by the high dose ICT treatment.The low dose ICT treatment moderately attenuated the above changes induced by the unloading.Mechanistically,Wortmannin could abolish the above effects of ICT.CONCLUSION ICT requires participation of PI3K/Akt signaling to counteract skeletal muscle atrophy following mechanical unloading in a dose-dependent manner.
文摘Sorafenib remains the standard systemic treatment for advanced human hepatocellular carcinoma(HCC).However,the low response rate,high recurrence,and high progression limit the therapeutic efficacy.Therefore,a combination therapy strategy was advanced to strengthen the antitumor effects of sorafenib.In the present study,we aimed to evaluate whether icaritin could enhance the inhibitory effects of sorafenib on HCC cells and clarify the underlying mechanism.The cell viability was evaluated via MTT assay,and the synergistic inhibitory effects of sorafenib and icaritin were verified by calculating the combination index(CI).Their combined effects on cell proliferation or apoptosis were investigated using colony formation assay and flow cytometry.Mitochondrial membrane potential(MMP)was detected by flow cytometric assay.The protein expressions associated with the apoptotic pathway were determined by Western blotting analysis.The data demonstrated that sorafenib and icaritin exerted synergistic inhibitory effects on cell viability(CI<1).Icaritin enhanced the inhibitory effect of sorafenib on colony formation and sorafenib-induced apoptosis of HCC cells.We discovered a reduced level of antiapoptotic Bcl-2 and an elevated level of proapoptotic protein Bax when the cells were exposed to the combination.The effect of cleaved and activated PARP was also enhanced.Cleaved caspase-9 and cleaved caspase-3 were increased markedly in the combination group.Furthermore,the combination of icaritin and sorafenib significantly increased the loss of MMP compared with the single treatment group and induced the release of cytochrome c from the mitochondria to the cytosol.These findings indicated that icaritin could enhance sorafenib-induced cytotoxicity and trigger sorafenib-induced apoptosis through a mitochondria-dependent pathway.
文摘The authors would like to apologize that,due to their neglect,some mistakes in the"2.Materials and methods"section need to be corrected.(1)The unit of DMAPP and flavonoid substrates in the“2.3.Enzymatic assays of prenyltransferases"section was not correctly written.The original description“200 mmol/L dimethylallyl pyrophosphate(DMAPP),200 mmol/L naringenin,kaempferol,or kaempferide as substrate"should be corrected to“200μmol/L dimethylallyl pyrophosphate(DMAPP),200μmol/L naringenin,kaempferol,or kaempferide as substrate"(2)The concentration of acetate acid in“2.8.Chemical analysis"was not correctly written.The original description“0.01%acetate acid(A)"should be corrected to“0.1%acetate acid(A)".The authors declare that these corrections do not change the results or conclusions of this paper.The authors sincerely apologize for this omission.
基金financially supported by the National Natural Science Foundation of China(Grant No.81770580)open funding from the Sichuan Provincial Key Laboratory of Radiation Oncology(Grant No.2020FSZLX-02).
文摘Background and Aims:Lipid accumulation is the major characteristic of non-alcoholic fatty liver disease,the prevalence of which continues to rise.We aimed to investigate the effects and mechanisms of icaritin on lipid accumulation.Methods:Cells were treated with icaritin at 0.7,2.2,6.7,or 20μM for 24 h.The effects on lipid accumulation in L02 and Huh-7 cells were detected by Bodipy and oil red O staining,respectively.Mitochondria biogenesis of L02 cells was detected by MitoTracker Orange staining.Glucose uptake and adenosine triphosphate content of 3T3-L1 adipocytes and C2C12 myotubes were detected.The expression levels of proteins in the adenosine 5′-monophosphate-activated protein kinase(AMPK)signaling pathway,biomarkers of autophagy,and mitochondria biogenesis were measured by western blotting.LC3 puncta were detected by immunofluorescence.Results:Icaritin significantly attenuated lipid accumulation in L02 and Huh-7 cells and boosted the mitochondria biogenesis of L02 cells.Icaritin enhanced glucose uptake,decreased adenosine triphosphate content,and activated the AMPK signaling pathway in 3T3-L1 adipocytes and C2C12 myotubes.Icaritin boosted autophagy and also enhanced the initiation of autophagic flux in 3T3-L1 preadipocytes and C2C12 myoblasts.However,icaritin decreased autophagy and promoted mitochondria biogenesis in 3T3-L1 adipocytes and C2C12 myotubes.Conclusions:Icaritin attenuates lipid accumulation by increasing energy expenditure and regulating autophagy by activating the AMPK pathway.
基金supported by the National Natural Science Foundation of China(No.30973944/H2805)the foundation for high-level talent on six areas of Jiangsu province.
文摘This study focuses on the preparation and enzymic hydrolysis of an icariin/bcyclodextrin inclusion complex to efficiently generate icaritin.The physical characteristics of the inclusion complex were evaluated by differential scanning calorimetry(DSC).Enzymatic hydrolysis was optimized for the conversion of icariin/b-cyclodextrin complex to icaritin by Box–Behnken statistical design.The inclusion complex formulation increased the solubility of icariin approximately 17-fold,from 29.2 to 513.5 mg/mL at 60℃.The optimum conditions were predicted by Box–Behnken statistical design as follows:60℃,pH 7.0,the ratio of enzyme/substrate(1:1.1)and reaction time 7 h.Under the optimal conditions the conversion of icariin was 97.91%and the reaction time was decreased by 68%compared with that without b-CD inclusion.Product analysis by melting point,ESI-MS,UV,IR,1H NMR and 13C NMR confirmed the authenticity of icaritin with a purity of 99.3%and a yield of 473 mg of icaritin from 1.1 g icariin.
基金financially supported by the Natural Foundation of Shandong Province (ZR2015QL002)
文摘Objective： To investigate the protective effects of icaritin （ICT）, one of the active ingredients in Epimedii Folium, on mouse model of cerebral ischemia-reperfusion （I/R） in vivo. Methods： ICR mice were subjected to an I h transient middle cerebral artery occlusion （MCAO） and fol- lowed by 24 h of reperfusion. Neurological deficits, infarct volume, brain edema and survive rate were measured, respectively. The levels of brain IL-1β, TNF-a, ROS and DNA-binding activity of NF-KB p65 were measured by ELISA kits. The levels of malondialdehyde （MDA） and activities of superoxide dismu- tase （SOD） were detected by spectrophotometry, and the release of nitric oxide （NO） were detected by Griess kit. Results： ICT markedly reduced the neurological deficit scores, brain edema, infarct volume and increased the survival rate of the cerebral I/R mice. The expression of IL-Iβ, TNF-α, NO, MDA and DNA-binding activity of NF-KB p65 were significantly inhibited by ICT, while the activity of SOD were up-regulated at the same time. Conclusion： ICT possessed significant neuroprotective effects in cerebral I/R mice, which might be related to prevent neuroinflammatory and oxidative damage.
基金EU-China Scientific and Technological Cooperation(1108)National Mega-project for Innovative Drugs(2012ZX09301002-001)Macao Science and Technology Development Fund(No.044/2011/A2)
文摘Objective Icaritin is the main aglycone and also active intestinal metabolite of prenylflavonoids from the Chinese materia medica Epimedii Herba. Modern pharmacological studies have demonstrated that icaritin has a wide range of biological activities. However, its metabolites and biotransformation pathways have not yet been comprehensively investigated. The present study aims to identify icaritin metabolites in rats by using a sensitive and effective LC-MS/MS method. Methods The plasma and urine samples of rats were collected before （blank） and after oral administration of icaritin, and subjected to liquid-liquid extraction with ethyl acetate. The full-scan LC-MS chromatograms of the plasma and urine samples were compared with those of blank samples to detect the possible metabolites, which were later detected by their product ion spectra. Results A total of 23 metabolites were identified, and conjugated icaritins produced by glucuronidation, glycosylation, and sulfation were its major metabolites. Minor demethylation, hydrogenation, and oxidation metabolites were also found. Conclusion Phase II metabolism is the main metabolic pathway of icaritin.
基金grants from National Natural Science Foundation of China(No.82373222,No.82202970)Leading Talent Program by Shanghai Municipal Health Commission(2022LJ008,China)+1 种基金Shanghai Shenkang Hospital Development Center(SHDC12021104,China)Science and Technology Commission of Shanghai Municipality(22Y21900200&22YF1404300,China).
文摘Tumor relapse and metastasis are the major causes of mortality associated with urothelial cancer.In the tumor microenvironment,negative regulatory molecules and various immune cell subtypes suppress antitumor immunity.The inflammatory microenvironment,associated with neutrophils and neutrophil extracellular traps(NETs),promotes tumor metastasis.However,no drugs are currently available to specifically inhibit neutrophils and NETs.In this study,we first demonstrated that icaritin(ICT),a Chinese herbal remedy that is a first-line treatment for advanced and incurable hepatocellular carcinoma,reduces NETs caused by suicidal NETosis and prevents neutrophil infiltration in the tumor microenvironment.Mechanistically,ICT binds to and inhibits the expression of PADI2 in neutrophils,thereby suppressing PADI2-mediated histone citrullination.Moreover,ICT inhibits ROS generation,suppresses the MAPK signaling pathway,and inhibits NET-induced tumor metastasis.Simultaneously,ICT inhibits tumoral PADI2-mediated histone citrullination,which consequently suppresses the transcription of neutrophil-recruiting genes such as GM-CSF and IL-6.The downregulation of IL-6 expression,in turn,forms a regulatory feedback loop through the JAK2/STAT3/IL-6 axis.Through a retrospective study of clinical samples,we found a correlation between neutrophils,NETs,UCa prognosis,and immune evasion.Combining ICT with immune checkpoint inhibitors may have synergistic effects.In summary,our study demonstrated that ICT could be a novel inhibitor of NETs and a novel UCa treatment.
基金supported by the National Natural Science Foundation of China (NSFC) [No. 81430095]
文摘Objective: To identify the in vivo metabolites of icaritin and speculate its metabolic profiling in rats.Methods: The plasma, bile, urine, and feces of rats were collected after orally administration of icaritin at a dose of 100 mg/kg and detected by an ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC/Q-TOF-MS/MS) in both positive and negative modes.The data of treated and control groups were compared and analyzed with the aid of Metabolynx XS software.Results: A total of 25 metabolites were identified in the biosamples, and 14 of them were reported for the first time to our knowledge.Conclusion: The main metabolite types of icaritin in rats were glucuronide conjugation, methylation, hydroxylation, reduction, and acetylation.
基金supported by the National Basic Research Program of China(Nos.2021YFA1201404,and 2019YFA0210103)the National Natural Science Foundation of China(Nos.32271413,and 82272492)+1 种基金Natural Science Foundation of Jiangsu Province(No.BK20232023)Science program of Jiangsu Province Administration for Market Regulation(No.KJ2024010).
文摘Bone tissue engineering provides a promising strategy for the treatment of bone defects.Nonetheless,the clinical utilization of biomaterial-based scaffolds is constrained by their inadequate mechanical strength and absence of osteo-inductive properties.Here,we proposed to endow nano-scaffold(NS)constructed by coaxial electrospinning technique with enhanced osteogenic bioactivities and mechanical properties by incorporating biocompatible magnetic iron oxide nanoparticles(IONPs)and icaritin(ICA).Four types of nano-scaffolds(NS,ICA@NS,NS-IONPs and ICA@NS-IONPs)were prepared.The incorporation of ICA and IONPs minimally impact their surface morphological and chemical properties.IONPs enhanced the mechanical properties of NS scaffolds,including hardness,tensile strength,and elastic modulus.In vitro assessments demonstrated that ICA@NS-IONPs exhibited enhanced osteogenic bioactivities towards mouse calvarial pre-osteoblast cell line MC3T3-E1 as evidenced by detecting the alkaline phosphatase(ALP)activity level,expressions of osteogenesis-related genes and proteins as well as mineralized nodule formation.Mechanistic investigations revealed that MEK/ERK(MAP kinase-ERK kinase(MEK)/extracellularsignal-regulated kinase(ERK))signaling pathway could offer a plausible explanation for the osteogenic differentiation of MC3T3-E1 cells induced by ICA@NS-IONPs.Furthermore,the implantation of nano-scaffolds in rat skull defects exhibited a substantial improvement in in vivo bone regeneration.Therefore,IONPs and ICA incorporated coaxial electrospinning nano-scaffolds present a novel strategy for the optimization of scaffolds for bone tissue engineering.
文摘中枢神经系统疾病(central nervous system diseases,CNSD)已成为日益严峻的全球性健康难题。中医药在CNSD方面展现独特的疗效与丰富的临床实践积累。淫羊藿Epimedii Folium作为一味传统中药,通过多靶点调控、多通路干预及多途径作用机制在神经系统疾病的治疗中发挥重要作用。随着淫羊藿药理作用的不断深入研究,发现其活性成分淫羊藿苷、淫羊藿素、淫羊藿次苷Ⅱ等是治疗CNSD的主要物质基础。淫羊藿活性成分可通过抑制炎症反应与氧化应激、抑制β-淀粉样蛋白沉积与微管相关蛋白异常磷酸化、抑制细胞凋亡、调节细胞自噬、改善神经元功能、调节神经递质等途径在阿尔茨海默病、帕金森病、癫痫、多发性硬化、脑缺血再灌注损伤、焦虑障碍、抑郁症、精神分裂症等CNSD中发挥作用。通过总结淫羊藿活性成分及其复方干预CNSD的作用机制,为进一步开发药物与临床运用奠定理论基础,为淫羊藿在CNSD研究中提供新的治疗方案和思路。
