Icariin accelerates bone regeneration by inducing osteogenesisangiogenesis coupling in rats with type 1 diabetes mellitus

BACKGROUND Icariin(ICA),a natural flavonoid compound monomer,has multiple pharmacological activities.However,its effect on bone defect in the context of type 1 diabetes mellitus(T1DM)has not yet been examined.AIM To explore the role and potential mechanism of ICA on bone defect in the context of T1DM.METHODS The effects of ICA on osteogenesis and angiogenesis were evaluated by alkaline phosphatase staining,alizarin red S staining,quantitative real-time polymerase chain reaction,Western blot,and immunofluorescence.Angiogenesis-related assays were conducted to investigate the relationship between osteogenesis and angiogenesis.A bone defect model was established in T1DM rats.The model rats were then treated with ICA or placebo and micron-scale computed tomography,histomorphometry,histology,and sequential fluorescent labeling were used to evaluate the effect of ICA on bone formation in the defect area.RESULTS ICA promoted bone marrow mesenchymal stem cell(BMSC)proliferation and osteogenic differentiation.The ICA treated-BMSCs showed higher expression levels of osteogenesis-related markers(alkaline phosphatase and osteocalcin)and angiogenesis-related markers(vascular endothelial growth factor A and platelet endothelial cell adhesion molecule 1)compared to the untreated group.ICA was also found to induce osteogenesis-angiogenesis coupling of BMSCs.In the bone defect model T1DM rats,ICA facilitated bone formation and CD31hiEMCNhi type H-positive capillary formation.Lastly,ICA effectively accelerated the rate of bone formation in the defect area.CONCLUSION ICA was able to accelerate bone regeneration in a T1DM rat model by inducing osteogenesis-angiogenesis coupling of BMSCs.

Icariin plus curcumol enhances autophagy through the mTOR pathway and promotes cathepsin B-mediated pyroptosis of prostate cancer cells

Objective:To examine the effect of icariin plus curcumol on prostate cancer cells PC3 and elucidate the underlying mechanisms.Methods:We employed the Cell Counting Kit 8 assay and colony formation assay to assess cell viability and proliferation.Autophagy expression was analyzed using monodansylcadaverine staining.Immunofluorescence and Western blot analyses were used to evaluate protein expressions related to autophagy,pyroptosis,and the mTOR pathway.Cellular damage was examined using the lactate dehydrogenase assay.Moreover,cathepsin B and NLRP3 were detected by co-immunoprecipitation.Results:Icariin plus curcumol led to a decrease in PC3 cell proliferation and an enhancement of autophagy.The levels of LC3-Ⅱ/LC3-Ⅰand beclin-1 were increased,while the levels of p62 and mTOR were decreased after treatment with icariin plus curcumol.These changes were reversed upon overexpression of mTOR.Furthermore,3-methyladenine resulted in a decrease in inflammatory cytokines,pyroptosis-related protein levels,and lactate dehydrogenase concentration,compared to the icariin plus curcumol group.Inhibiting cathepsin B reversed the regulatory effects of icariin plus curcumol.Conclusions:Icariin plus curcumol demonstrates great potential as a therapeutic agent for castration-resistant prostate cancer by enhancing autophagy via the mTOR pathway and promoting pyroptosis mediated by cathepsin B.These findings provide valuable insights into the molecular mechanisms underlying the therapeutic potential of icariin and curcumol for prostate cancer treatment.

Icariin ameliorates viral myocarditis by inhibiting TLR4-mediated ferroptosis

Objective:To explore the mechanism by which icariin alleviates viral myocarditis.Methods:CVB3-induced cardiomyocytes were used as an in vitro model of viral myocarditis to assess the effects of icariin treatment on cell viability,inflammation,and apoptosis.Moreover,the effects of icariin on ferroptosis and TLR4 signaling were assessed.After AC16 cells were transfected with TLR4 overexpression plasmids,the role of TLR4 in mediating the regulatory effect of icariin in viral myocarditis was investigated.Results:Icariin significantly elevated cell viability and reduced inflammatory factors TNF-α,IL-1β,IL-6,and IL-18.Flow cytometry revealed that icariin decreased apoptosis rate,and the protein expression of Bax and cleaved caspase 3 and 9 in CVB3-induced cardiomyocytes.Additionally,it suppressed ferroptosis including lipid peroxidation and ferrous ion,as well as the TLR4 signaling.However,TLR4 overexpression abrogated the modulatory effects of icariin.Conclusions:Icariin mitigates CVB3-induced myocardial injury by inhibiting TLR4-mediated ferroptosis.Further animal study is needed to verify its efficacy.

Icariin Ameliorates D-galactose-induced Cell Injury in Neuron-like PC12 Cells by Inhibiting MPTP Opening

Objective Icariin(ICA)has a good neuroprotective effect and can upregulate neuronal basal autophagy in naturally aging rats.Mitochondrial dysfunction is associated with brain aging-related neurodegenerative diseases.Abnormal opening of the mitochondrial permeability transition pore(mPTP)is a crucial factor in mitochondrial dysfunction and is associated with excessive autophagy.This study aimed to explore that ICA protects against neuronal injury by blocking the mPTP opening and down-regulating autophagy levels in a D-galactose(D-gal)-induced cell injury model.Methods A cell model of neuronal injury was established in rat pheochromocytoma cells(PC12 cells)treated with 200 mmol/L D-gal for 48 h.In this cell model,PC12 cells were pre-treated with different concentrations of ICA for 24 h.MTT was used to detect cell viability.Senescence associatedβ-galactosidase(SA-β-Gal)staining was used to observe cell senescence.Western blot analysis was performed to detect the expression levels of a senescence-related protein(p21),autophagy markers(LC3B,p62,Atg7,Atg5 and Beclin 1),mitochondrial fission and fusion-related proteins(Drp1,Mfn2 and Opa1),and mitophagy markers(Pink1 and Parkin).The changes of autophagic flow were detected by using mRFP-GFP-LC3 adenovirus.The intracellular ultrastructure was observed by transmission electron microscopy.Immunofluorescence was used to detect mPTP,mitochondrial membrane potential(MMP),mitochondrial reactive oxygen species(mtROS)and ROS levels.ROS and apoptosis levels were detected by flow cytometry.Results D-gal treatment significantly decreased the viability of PC12 cells,and markedly increased the SA-β-Gal positive cells as compared to the control group.With the D-gal stimulation,the expression of p21 was significantly up-regulated.Furthermore,D-gal stimulation resulted in an elevated LC3B II/I ratio and decreased p62 expression.Meanwhile,autophagosomes and autolysosomes were significantly increased,indicating abnormal activation of autophagy levels.In addition,in this D-gal-induced model of cell injury,the mPTP was abnormally open,the ROS generation was continuously increased,the MMP was gradually decreased,and the apoptosis was increased.ICA effectively improved mitochondrial dysfunction to protect against D-gal-induced cell injury and apoptosis.It strongly inhibited excessive autophagy by blocking the opening of the mPTP.Cotreatment with ICA and an mPTP inhibitor(cyclosporin A)did not ameliorate mitochondrial dysfunction.However,the protective effects were attenuated by cotreatment with ICA and an mPTP activator(lonidamine).Conclusion ICA inhibits the activation of excessive autophagy and thus improves mitochondrial dysfunction by blocking the mPTP opening.

Bibliometric analysis of 100 most-cited papers on Icariin

Icariin is the most prevalent component of the medicinal herb Herba Epimedii.Icariin exhibits many medicinal properties,including anti-cancer impact and osteoprotective and neuroprotective effects.The goal of this study was to use bibliometric analysis to find and describe the top 100 papers about Icariin that had received the most citations.The Science Citation Index-Expanded(SCI-E)of the Web of Science Core Collection was used to find publications on Icariin(WoSCC).Descriptive analysis was conducted using VOSviewer software.There were 1473 articles about Icariin in all.The top 100 papers were published between 1996 and 2024 and received citations in the range of 55 to 390.The country that has contributed the most to Icariin research is China(84).The most productive institution was Fudan University.The most published journal was Phytomedicine.The research hotspots of Icariin mainly focus on the following aspects:research on Icariin treatment of sex hormone-related osteoporosis and erectile function;The effect of Icariin on cells by regulating oxidative stress,apoptosis,and proliferation;the mechanism of Icariin in the treatment of cancer;the neuroprotective effect of Icariin in central nervous diseases,such as Alzheimer's disease,Parkinson's disease,and depression.Future research should focus on further elucidating Icariin's anti-tumor effects,its application in cartilage tissue engineering and orthopedic biomaterials,and developing novel drug delivery systems to enhance its bioavailability.This research contributed essential knowledge to the study of Icariin.These results may be used in new study areas and to direct drug development.

Analysis of the Antidepressant Effects and Mechanisms of Icariin II Based on The GABAergic Nervous System

This article summarized the research progress on the antidepressant mechanism of icariin II,mainly elaborating on its mechanism from five aspects:GABAergic nervous system,inflammatory response,oxidative stress,neurotrophic factors,and neurotransmitters in the brain.Its clinical application value was further explored to provide a theoretical basis for the development and utilization of icariin II in treating depression.

Icariin ameliorates memory deficits through regulating brain insulin signaling and glucose transporters in 3×Tg-AD mice

Icariin,a major prenylated flavonoid found in Epimedium spp.,is a bioactive constituent of Herba Epimedii and has been shown to exert neuroprotective effects in experimental models of Alzheimer’s disease.In this study,we investigated the neuroprotective mechanism of icariin in an APP/PS1/Tau triple-transgenic mouse model of Alzheimer’s disease.We performed behavioral tests,pathological examination,and western blot assay,and found that memory deficits of the model mice were obviously improved,neuronal and synaptic damage in the cerebral cortex was substantially mitigated,and amyloid-βaccumulation and tau hyperphosphorylation were considerably reduced after 5 months of intragastric administration of icariin at a dose of 60 mg/kg body weight per day.Furthermore,deficits of proteins in the insulin signaling pathway and their phosphorylation levels were significantly reversed,including the insulin receptor,insulin receptor substrate 1,phosphatidylinositol-3-kinase,protein kinase B,and glycogen synthase kinase 3β,and the levels of glucose transporter 1 and 3 were markedly increased.These findings suggest that icariin can improve learning and memory impairments in the mouse model of Alzheimer’s disease by regulating brain insulin signaling and glucose transporters,which lays the foundation for potential clinical application of icariin in the prevention and treatment of Alzheimer’s disease.

Bone mesenchymal stem cell-derived exosomes involved co-delivery and synergism effect with icariin via mussel-inspired multifunctional hydrogel for cartilage protection

Mesenchymal stem cells(MSC)are particularly effective in promoting cartilage regeneration due to their immunomodulatory,anti-inflammatory and regenerative repair functions of tissues and organs.Meanwhile,the intra-articular delivery and synergy with other therapeutic drugs have been the key issues driving their further application.We report a mussel-inspired multifunctional hydrogel system,which could achieve co-delivery and synergism effect of MSC-derived exosomes(Exos)with icariin(ICA).The ICA and Exos co-delivered articular cavity injection system are expected to retain in the joint cavity and promote cartilage regeneration,due to the thermosensitive,self-healing and adhesion properties of the mussel-inspired multifunctional hydrogel.The experimental results proved that Exos enhanced the cellular uptake of ICA by more than 2-fold evenly,and the synergism of Exos and ICA efficiently improve the cell proliferation and migration.After synergic treatment,the content of matrix metalloproteinase 13 in the supernatant and intracellular decreased by 47%and 59%,respectively.In vivo study,ICA-loaded Exos exhibited prolonged retention behavior bymultifunctional hydrogel delivery,thus displayed an increased cartilage protection.In the model of osteoarthritis,co-delivery hydrogel system relieved the cartilage recession,ensuring appropriate cartilage thickness.

Potential role of microRNA-503 in Icariin-mediated prevention of high glucose-induced endoplasmic reticulum stress

BACKGROUND Dysregulated microRNA(miRNA)is crucial in the progression of diabetic nephropathy(DN).AIM To investigate the potential molecular mechanism of Icariin(ICA)in regulating endoplasmic reticulum(ER)stress-mediated apoptosis in high glucose(HG)-induced primary rat kidney cells(PRKs),with emphasis on the role of miR-503 and sirtuin 4(SIRT4)in this process.METHODS Single intraperitoneal injection of streptozotocin(65 mg/kg)in Sprague-Dawley rats induce DN in the in vivo hyperglycemic model.Glucose-treated PRKs were used as an in vitro HG model.An immunofluorescence assay identified isolated PRKs.Cell Counting Kit-8 and flow cytometry analyzed the effect of ICA treatment on cell viability and apoptosis,respectively.Real-time quantitative polymerase chain reaction and western blot analyzed the levels of ER stressrelated proteins.Dual luciferase analysis of miR-503 binding to downstream SIRT4 was performed.RESULTS ICA treatment alleviated the upregulated miR-503 expression in vivo(DN)and in vitro(HG).Mechanistically,ICA reduced HG-induced miR-503 overexpression,thereby counteracting its function in downregulating SIRT4 levels.ICA regulated the miR-503/SIRT4 axis and subsequent ER stress to alleviate HG-induced PRKs injury.CONCLUSION ICA reduced HG-mediated inhibition of cell viability,promotion of apoptosis,and ER stress in PRKs.These effects involved regulation of the miR-503/SIRT4 axis.These findings indicate the potential of ICA to treat DN,and implicate miR-503 as a viable target for therapeutic interventions in DN.

淫羊藿苷对抑郁症模型小胶质细胞表型转化的调控作用

探索淫羊藿苷抗抑郁的作用机制,将SPF级KM小鼠分为空白组、模型组、盐酸氟西汀组(10 mg/kg)、淫羊藿苷高(50 mg/kg)、低(25 mg/kg)剂量组。除空白组外,其余各组小鼠采用56 d慢性不可预知温和应激建立抑郁症模型。造模第1 d开始灌胃给予各组小鼠相应药物,连续56 d。于给药后第53~56 d,进行行为学测试。末次给药后,取材。酶联免疫吸附剂实验(enzyme linked immunosorbent assay,ELISA)检测脑组织匀浆白细胞介素6(interleukin-6,IL-6)、白细胞介素10(interleukin-10,IL-10)、五羟色胺(5-hydroxytryptamine,5-HT)、多巴胺(dopamine,DA)、去甲肾上腺素(norepinephrine,NE)水平;实时荧光定量PCR(quantitative real-time PCR,RT-PCR)检测脑组织中IL-6、IL-10、诱导性一氧化氮合酶(inducible nitric oxide synthase,iNOS)、分化簇206(cluster of differentiation 206,CD206)mRNA表达;蛋白免疫印迹(Western blot)检测脑组织iNOS、CD206蛋白表达。体外培养小鼠小胶质细胞(BV-2),采用细胞增殖与毒性检测试剂盒(Cell Counting Kit-8,CCK-8)法检测淫羊藿苷的细胞毒性。BV-2细胞暴露于100μg/mL脂多糖及15μg/mL、25μg/mL的淫羊藿苷24 h后,收集细胞。免疫荧光技术检测细胞iNOS、CD206蛋白表达。研究结果表明,慢性不可预知温和应激致抑郁症模型制备成功,淫羊藿苷可降低抑郁症小鼠脑组织IL-6水平及IL-6、iNOS mRNA表达,升高脑组织IL-10、5-HT、DA、NE水平及IL-10、CD206 mRNA表达;降低脑组织iNOS蛋白,升高CD206蛋白表达;改善模型小鼠抑郁症样行为。淫羊藿苷可降低LPS刺激的BV-2细胞iNOS蛋白表达,升高CD206蛋白表达。结果表明,淫羊藿苷抑制小胶质细胞M1表型转化,促进M2表型转化,从而改善模型小鼠抑郁样行为。

淫羊藿苷调控mTOR/Akt/CREB通路对高糖诱导的足细胞自噬及凋亡的影响

目的 探讨淫羊藿苷对高糖诱导的足细胞自噬、凋亡及哺乳动物雷帕霉素靶蛋白(mTOR)/丝氨酸苏氨酸蛋白激酶(Akt)/环磷酸腺苷反应元件结合蛋白(CREB)通路的影响。方法 将小鼠足细胞MPC5分为5组:正常对照组(5.5 mmol·L^(-1)葡萄糖)、高糖组(30 mmol·L^(-1)葡萄糖)、淫羊藿苷组(30 mmol·L^(-1)葡萄糖+5μmol·L^(-1)淫羊藿苷)、GDC-0349组(30 mmol·L^(-1)葡萄糖+50μmol·L^(-1)GDC-0349)、淫羊藿苷+GDC-0349组(30 mmol·L^(-1)葡萄糖+5μmol·L^(-1)淫羊藿苷+50μmol·L^(-1)GDC-0349)。培养48 h后,噻唑蓝法检测MPC5细胞活力;吖啶橙染色观察MPC5细胞自噬情况;流式细胞术检测MPC5细胞凋亡;蛋白印迹法检测MPC5细胞自噬[微管相关蛋白1轻链3(LC3)Ⅱ、LC3Ⅰ、自噬相关蛋白(Beclin-1)]、凋亡[Bcl-2相关X蛋白(Bax)、B淋巴细胞瘤-2(Bcl-2)]和mTOR/Akt/CREB通路相关蛋白的表达。结果 与正常对照组比较,高糖组MPC5细胞活力、Bcl-2、磷酸化mTOR(p-mTOR)/mTOR、磷酸化Akt(p-Akt)/Akt、磷酸化CREB(p-CREB)/CREB蛋白表达水平显著降低(P<0.05),自噬能力增强,自噬体表现出橙色荧光,细胞凋亡率、LC3Ⅱ/LC3Ⅰ、Beclin-1、Bax蛋白表达水平显著升高(P<0.05)。与高糖组比较,淫羊藿苷组MPC5细胞活力、LC3Ⅱ/LC3Ⅰ、Beclin-1、Bcl-2、p-mTOR/mTOR、p-Akt/Akt、p-CREB/CREB蛋白表达水平显著升高,自噬能力进一步增强,自噬体数量增多,自噬体呈现出砖红色荧光(P<0.05),细胞凋亡率、Bax蛋白表达水平显著降低(P<0.05);GDC-0349组MPC5细胞活力、LC3Ⅱ/LC3Ⅰ、Beclin-1、Bcl-2、p-mTOR/mTOR、p-Akt/Akt、p-CREB/CREB蛋白表达水平显著降低,自噬能力减弱,自噬体数量减少,自噬体表现出橙色荧光(P<0.05),细胞凋亡率、Bax蛋白表达水平显著升高(P<0.05);淫羊藿苷+GDC-0349可逆转淫羊藿苷对高糖诱导MPC5细胞的作用效果(P<0.05)。结论 淫羊藿苷通过激活mTOR/Akt/CREB通路促进高糖诱导的足细胞自噬抑制细胞凋亡。

Exploring the mechanism of icariin in regulat⁃ing cardiac microvascular endothelial cells based on network pharmacology,molecular docking and in vitro experiments

OBJECTIVE To investigate the regulatory effects of icariin(ICA)on cardiac micro⁃vascular endothelial cells(CMEC)after oxygenglucose deprivation reperfusion(OGD/R)injury.METHODS CMEC were subjected to OGD/R treatment to construct a myocardial ischemiareperfusion model,and were divided into normal,model,low(10μmol·L^(-1)),medium(20μmol·L^(-1))and high(40μmol·L^(-1))ICA group,and high ICA+inhibitor group(40μmol·L^(-1)+20 nmol·L^(-1)).CCK-8 assay was used to assess the protective ability of ICA against CMEC,and cell migration assay and tube-formation assay were used to detect the migration and generation ability of CMEC.The TCMSP database,Swiss-Target database and literature mining methods were used to col⁃lect ICA-related targets,the GeneCards data⁃base was used to collect target genes related to myocardial ischemia/reperfusion,and Cytoscape 3.8.0 software was used to construct a"drug-tar⁃get-disease"network.The potential targets were imported into STRING 11.5 database to obtain the PPI network.GO and KEGG enrichment analyses were performed on the potential targets using the DAVID database.Molecular docking was performed using AutoDock-vina 1.1.2 soft⁃ware.Western blot detected the expression of related proteins.RESULTS After CMEC was subjected to OGD/R treatment,ICA had a protec⁃tive effect at 10^(-1)60μmol·L^(-1);the results of the cell migration assay showed that each group of ICA could promote the migratory effect of CMEC(P<0.01,P<0.01);and the results of tube-for⁃mation assay showed that each group of ICA could significantly promote the generation of branches(P<0.01)and the capillary length exten⁃sion(P<0.05).Network pharmacology collected a total of 23 ICA action targets,1500 disease tar⁃gets and 12 key targets.GO function enrichment analysis found 85 results.KEGG pathway enrich⁃ment analysis found 53 results,involving AGERAGE signaling pathway,sphingolipid signaling pathway and VEGF signaling pathway.Molecu⁃lar docking results showed that ICA had better binding with core targets PRKCB,PRKCA and PTGS2.Western blot results showed that ICA could regulate the expression of PRKCB,PRKCA and PTGS2 proteins.The results of cell migra⁃tion assay,tube-formation assay and protein expression were reversed after addition of PKC inhibitor.CONCLUSION The potential mecha⁃nism of action of ICA against myocardial isch⁃emia-reperfusion injury may be related to the reg⁃ulation of processes such as CMEC migration and angiogenesis,and it functions through the key target gene PKC.

基于不同炮制方法的淫羊藿黄酮类成分含量研究

试验旨在寻求羊脂油炮制淫羊藿的内在机理,以甘肃产淫羊藿为研究对象,用羊脂油、牛脂油、猪脂油进行炮制,分别提取有效成分,利用紫外分光光度法和高效液相色谱法进行有效成分含量测定,比较不同炮制品和生品总黄酮、淫羊藿苷、宝藿苷Ⅰ和朝藿定C的含量。结果显示:各炮制品总黄酮含量均显著低于生品(6.84%),其中羊脂油炙含量(5.69%)显著高于其他炮制品;生品和羊、牛脂油炙炮制品中淫羊藿苷含量无显著差异,羊脂油炙炮制品为0.77%,牛脂油炙炮制品为0.70%,生品为0.70%,猪脂油炙炮制品中淫羊藿苷含量为0.50%,较生品和其他炮制品显著降低;羊脂油炙、牛脂油炙淫羊藿中宝藿苷Ⅰ含量无显著性差异,分别为0.08%和0.07%,但显著高于生品(0.05%)和猪脂油炙淫羊藿(0.04%);羊、牛、猪脂油炙炮制品中朝藿定C含量分别为0.29%、0.27%和0.24%,均显著低于生品(0.57%)。得出结论:淫羊藿最佳炮制辅料为羊脂油,其次为牛脂油。

淫羊藿苷对前列腺癌原位移植瘤小鼠肿瘤生长及细胞周期的影响

目的探讨淫羊藿苷对前列腺癌原位移植瘤SCID小鼠肿瘤生长、细胞周期、磷酸酶及张力蛋白同源物(phosphatase and tensin homolog,PTEN)基因表达的影响。方法40只雄性SCID小鼠,根据随机数字表法随机均分为模型对照组、低剂量组、中剂量组、高剂量组,每组10只。采用SCID小鼠前列腺腺体背外侧包膜内注射人前列腺癌LNCaP细胞株悬液的方法构建前列腺癌原位移植瘤小鼠模型。造模2周后,模型对照组予以0.9%氯化钠注射液,低剂量组予以10 mg/kg淫羊藿苷,中剂量组予以40 mg/kg淫羊藿苷,高剂量组予以80 mg/kg淫羊藿苷。灌胃处理1次/d,治疗时间为5周。在灌胃给药治疗前及治疗5周后,分别取各组小鼠5只,对比各组肿瘤质量、肿瘤体积、肿瘤抑制率。采用流式细胞学方法检测LNCaP细胞周期,计算各周期比值。采用实时定量PCR检测前列腺肿瘤组织中PTEN mRNA相对含量。结果治疗后,模型对照组肿瘤质量、肿瘤体积较治疗前明显增加(P<0.05);中剂量组及高剂量组肿瘤质量、肿瘤体积较治疗前明显降低(P<0.05),且显著低于模型对照组(P<0.05)。中剂量组及高剂量组肿瘤抑制率显著高于低剂量组(P<0.05)。中剂量组及高剂量组肿瘤细胞S期较治疗前明显增加(P<0.05),且显著高于模型对照组(P<0.05);中剂量组及高剂量组肿瘤细胞G_(0)/G_(1)期较治疗前明显减少(P<0.05),且显著低于模型对照组(P<0.05)。中剂量组及高剂量组PTEN mRNA相对含量较治疗前明显增加(P<0.05),且显著高于模型对照组(P<0.05)。结论淫羊藿苷可能通过上调PTEN表达、改变细胞周期分布来抑制前列腺癌LNCaP细胞增殖,从而有效抑制前列腺癌原位移植瘤SCID小鼠肿瘤生长。

淫羊藿活性成分抗卵巢癌作用机制研究进展

卵巢癌是女性生殖系统三大癌症之一,其病死率位居妇科恶性肿瘤之首。淫羊藿具有补肾阳、强筋骨、祛风湿的功效,临床常用于生殖系统疾病的治疗。本文通过综述国内外相关文献,发现淫羊藿主要活性成分淫羊藿苷、淫羊藿素和淫羊藿次苷Ⅱ,主要从促进细胞凋亡、影响细胞的侵袭和转移、阻滞细胞周期、诱导细胞自噬、促进细胞焦亡等5个方面发挥抗卵巢癌作用,可见淫羊藿可以通过多靶点、多途径治疗卵巢癌。

HPLC法同时测定驻春胶囊中4个成分的含量

目的:建立以高效液相色谱法(HPLC)同时测定驻春胶囊中异补骨脂素、补骨脂素、淫羊藿苷、蛇床子素含量的方法。方法:采用HPLC法,以异补骨脂素、补骨脂素、淫羊藿苷和蛇床子素作为指标成分,采用Agilent ZORBAX SB-C_(18)色谱柱(4.6 mm×250 mm,5μm);流动相为甲醇-水,梯度洗脱,体积流量为1.0 m·min^(-1);柱温为30℃,检测波长为254 nm,进样量为5μL。结果:异补骨脂素、补骨脂素、淫羊藿苷和蛇床子素质量浓度分别在0.0059~0.1770 mg/mL、0.0049~0.1470 mg/mL、0.0031~0.093 mg/mL和0.0048~0.1440 mg/mL范围内与峰面积呈良好的线性关系,其相关系数分别为0.9996、0.9995、0.9997和0.9995,平均加样回收率分别为101.45%、100.95%、100.46%和101.17%;RSD分别为2.19%、1.83%、1.71%和2.01%。结论:该分析方法适用于驻春胶囊中异补骨脂素、补骨脂素、淫羊藿苷和蛇床子素的含量测定。

淫羊藿苷在炎症环境下对MC3T3-E1细胞增殖分化的影响

背景:研究表明慢性根尖周炎是常见的炎症性骨破坏疾病之一,淫羊藿苷可以促进成骨分化、抑制骨吸收,可能对慢性根尖周炎引起的骨破坏起到保护作用。目的:探讨在脂多糖刺激的炎症环境下淫羊藿苷对MC3T3-E1细胞增殖分化的影响。方法:应用脂多糖刺激MC3T3-E1细胞建立体外炎症环境,采用CCK-8检测脂多糖最佳作用质量浓度及作用时间;CCK-8检测在1μg/mL脂多糖刺激下淫羊藿苷的最佳作用质量浓度;碱性磷酸酶染色、Real-time PCR及Western blot检测炎症环境下淫羊藿苷对MC3T3-E1细胞成骨分化的影响;Real-time PCR及Western blot检测炎症环境下淫羊藿苷对MC3T3-E1细胞炎症相关因子白细胞介素1β、白细胞介素6表达的影响。结果与结论:①CCK-8结果显示,淫羊藿苷的最佳作用质量浓度为0.1μg/mL;②在炎症环境下,淫羊藿苷增强了碱性磷酸酶的表达,促进了成骨细胞分化;③与脂多糖组相比,脂多糖+淫羊藿苷组成骨相关因子碱性磷酸酶、Runx2表达升高;④与脂多糖组相比,脂多糖+淫羊藿苷组炎症相关因子白细胞介素1β、白细胞介素6表达水平下降;⑤结果提示,脂多糖能导致MC3T3-E1细胞成骨分化能力减弱并加重炎症反应,淫羊藿苷对其具有保护作用。

具有病灶治愈功能的淫羊藿苷洗脱涂层修饰血管支架

目的开发一种ICA洗脱涂层,以聚乳酸-羟基乙酸共聚物(PLGA)为药物递送载体,实现靶向治疗。方法利用超声雾化喷涂技术在血管支架表面构建ICA洗脱涂层,利用傅里叶变换红外吸收光谱仪(FTIR)、水接触角(WCA)检测仪和场发射扫描电子显微镜(SEM)等表征手段对涂层的材料学性质进行检测;通过紫外分光光度计(UV-Vis)检测药物释放行为;通过体外溶血率和血小板的黏附与激活实验验证该涂层的血液相容性;通过体外细胞实验进一步评价该涂层的细胞相容性。结果研究结果显示,涂层构建成功,且ICA的装载并未对PLGA涂层的力学性能产生影响,同时实现了ICA体外至少28 d的持续释放。血液相容性的评价结果显示,该涂层有效减少了血小板的黏附和聚集,且该涂层能够抑制SMCs的异常增殖,促进ECs的黏附、增殖,有效调控巨噬细胞向组织修复型转变。结论ICA洗脱支架涂层显示出潜在的抗血栓、抗炎、促内皮和抑制平滑肌细胞异常增殖的特性,有望为心血管疾病患者提供一种安全和有效的治疗选择。

一种新型负载淫羊藿苷的普鲁兰纳米粒制备及其性能研究

目的 设计一种新型淫羊藿苷修饰的普鲁兰聚合物,通过化学连接和物理包埋的方式高效装载淫羊藿苷,制备一种具有缓释性的纳米球形载体,并对其性能进行初步探讨。方法 淫羊藿苷通过丁二酸酐作为连接臂接枝亲水性普鲁兰形成淫羊藿苷修饰的普鲁兰聚合物,通过傅里叶红外光谱对终产物鉴定。采用透析法制备负载淫羊藿苷的普鲁兰纳米粒。测量空白纳米粒和载药纳米粒的粒径和Zeta电位,利用透射电镜观察其形貌,并测定其在不同pH条件下的体外药物释放行为。结果 纳米粒具有规则的形态、均匀的尺寸。空白纳米粒电位为+2.23 mV,载药纳米粒电位为+3.98 mV;空白纳米粒平均粒径为122 nm,载药纳米粒粒径为190 nm。纳米粒能明显延长淫羊藿苷的缓释作用时间,在pH 6.8的酸性条件下更有利于淫羊藿苷的释放。结论 通过两亲性聚合物自组装行为,成功制备了尺寸均匀的纳米粒。通过聚合物化学连接方式丁二酸将中药单体淫羊藿苷连接普鲁兰制备为纳米递送系统,淫羊藿苷的普鲁兰纳米递送系统该系统能使疏水性药物成为纳米型的水溶性药物,药物具有缓释性,并在弱酸度环境下可以定向释放。

淫羊藿苷对四氯化碳诱导小鼠急性肝损伤的保护作用机制

背景:淫羊藿苷具有抗炎、抗氧化、免疫调节等功能,可能成为预防和治疗急性肝损伤的潜在药物。目的:分析淫羊藿苷对四氯化碳诱导小鼠急性肝损伤的保护作用及可能机制。方法:取32只昆明小鼠,采用随机数字表法分为正常组、模型组、淫羊藿苷低剂量组及淫羊藿苷高剂量组,每组8只。正常组、模型组灌胃给予10 mL/kg的生理盐水,淫羊藿苷低剂量组、淫羊藿苷高剂量组分别灌胃给予100,200 mg/kg的淫羊藿苷混悬液,1次/d,连续7 d。末次给药后,模型组、淫羊藿苷低剂量组及淫羊藿苷高剂量组通过腹腔注射四氯化碳建立急性肝损伤模型,造模24 h后,检测小鼠肝指数与血清中丙氨酸氨基转移酶、天冬氨酸氨基转移酶、肿瘤坏死因子α及白细胞介素6水平,肝组织中超氧化物歧化酶、谷胱甘肽过氧化物酶及丙二醛浓度,肝组织病理形态及肝细胞凋亡情况,以及肝组织中混葡萄糖调节蛋白78、内质网应激相关蛋白、混合谱系激酶域样蛋白及Caspase-3的蛋白表达。结果与结论:①与正常组比较,模型组小鼠肝指数与血清中丙氨酸氨基转移酶、天冬氨酸氨基转移酶、肿瘤坏死因子α及白细胞介素6水平均升高(P均<0.05);与模型组比较,淫羊藿苷低剂量组、淫羊藿苷高剂量组小鼠上述指标均降低(P均<0.05);②与正常组比较,模型组小鼠肝组织中超氧化物歧化酶、谷胱甘肽过氧化物酶活性降低(P均<0.05),丙二醛水平升高(P<0.05);与模型组比较,淫羊藿苷低剂量组、淫羊藿苷高剂量组小鼠超氧化物歧化酶、谷胱甘肽过氧化物酶活性升高(P均<0.05),丙二醛水平降低(P<0.05);③苏木精-伊红与TUNEL染色显示,模型组小鼠肝组织结构破坏严重,肝细胞大面积坏死,肝细胞凋亡率较高;淫羊藿苷低剂量组及淫羊藿苷高剂量组小鼠肝组织结构破坏程度及肝细胞坏死面积明显轻于模型组,肝细胞凋亡率低于模型组(P<0.05);④Western blot检测显示,与正常组比较,模型组小鼠肝组织内葡萄糖调节蛋白78、内质网应激相关蛋白、混合谱系激酶域样蛋白Caspase-3的蛋白表达升高(P均<0.05);与模型组比较,淫羊藿苷低剂量组、淫羊藿苷高剂量组小鼠肝组织内上述指标蛋白表达降低(P均<0.05);⑤结果表明,淫羊藿苷对四氯化碳诱导的急性肝损伤具有保护作用,其机制可能与抗炎、抗氧化、调节内质网应激及减轻程序性坏死有关。