肉苁蓉苯乙醇总苷脂质体对rrPDGF-BB诱导的肝星状细胞增殖的影响及作用机制研究  

Study on effect of liposomes from cistanche phenylethanoid glycosides on proliferation of rrPDGF-BB induced hepatic stellate cells and its action mechanism

作　　者：马晓婷[1] 张石蕾 王志强[1] 陈文龙 由淑萍[1] 刘涛[1] MA Xiaoting;ZHANG Shilei;WANG Zhiqiang;CHEN Wenlong;YOU Shuping;LIU Tao(College of Public Health,Xinjiang Medical University,Urumqi,Xinjiang 830011,China)

机构地区：[1]新疆医科大学公共卫生学院,乌鲁木齐830011

出　　处：《重庆医学》2019年第10期1630-1634,共5页Chongqing medicine

基　　金：国家自然科学基金项目(81560628)

摘　　要：目的探讨肉苁蓉苯乙醇总苷(CPhGs)脂质体对重组大鼠血小板衍生生长因子-BB(rrPDGFBB)诱导的大鼠肝星状细胞(HSC)增殖作用的影响及其抗肝纤维化的相关分子机制。方法取生长状态良好的对数生长期HSC用于实验,置于CPhGs脂质体浓度为117.79、58.90、29.45、14.72、7.36、3.68、1.84、0.92和0.46mg/L的完全培养液,求出半数抑制浓度(IC_(50));rrPDGF-BB刺激后,采用四甲基偶氮唑盐(MTT)法测定29.45、14.72、7.36mg/L CPhGs脂质体对HSC增殖的影响;采用实时荧光定量聚合酶链反应(qRT-PCR)检测α-平滑肌肌动蛋白(α-SMA)、Ⅰ型胶原、ATF-2mRNA的表达水平;采用Western blot法检测CPhGs脂质体对p-p38蛋白表达的影响。结果 (1)对细胞增殖的影响:对不同时间点结果分析显示,与Normal组比较,细胞培养48h时,rrPDGF-BB组HSC细胞明显增多;CPhGs脂质体各剂量组抑制rrPDGF-BB诱导的HSC增殖作用,培养48h效率最高;对不同浓度结果分析显示,与rrPDGF-BB组比较,不同浓度CPhGs脂质体组均可抑制rrPDGF-BB诱导的HSC-T6的增殖。(2)qRT-PCR结果:CPhGs脂质体各剂量组间比较,随CPhGs脂质体药物浓度增大,α-SMA、Ⅰ型胶原和ATF-2 mRNA水平显著下降,差异有统计学意义(P<0.05),其中以CPhGs脂质体29.45mg/L组抑制效果最明显。(3)Western blot分析结果:p-p38蛋白的表达水平在CPhGs脂质体不同浓度组表达均下降,差异有统计学意义(P<0.05);CPhGs脂质体各剂量组间比较,p-P38蛋白随CPhGs脂质体干预剂量增大,表达水平显著下降(P<0.05)。结论 CPhGs脂质体能抑制rrPDGF-BB诱导的大鼠HSC增殖,从而起到抗肝纤维化的作用。Objective To investigate the effect of cistanche phenylethanoid glycosides(CPhGs)liposomes on the proliferation of rat hepatic stellate cells(HSC)induced by platelet-derived growth factor-BB(rrPDGF-BB)and the related molecular mechanisms of its anti-hepatic fibrosis.Methods HSC in logarithmic growth phase were cultured in complete medium with CPhGs liposome concentrations of 117.79,58.90,29.45,14.72,7.36,3.68,1.84,0.92 and 0.46 mg/L.And the half maximal inhibitory concentration(IC50)was calculated.After rrPDGF-BB stimulation,the effects of liposome concentrations of 29.45,14.72,7.36 mg/L on HSC proliferation were detected by MTT method.Real-time fluorescence quantitative PCR(qRT-PCR)was used to detect the expression ofα-SMA,collagenⅠ and ATF-2 mRNA.Western blot was used to detect the influence of CPhGs liposome on the expression of p-p38 protein.Results(1)Effect on cell proliferation:compared with the normal group,HSC in the rrPDGF-BB group increased significantly after 48 hours of cell culture.CPhGs liposome inhibited the proliferation of HSC induced by rrPDGF-BB with the highest efficiency after 48 hours of culture.CPhGs liposome inhibited the proliferation of HSC at different concentrations compared with the rrPDGF-BB group,and CPhGs liposome at different concentrations all could inhibit the proliferation of HSC-6 induced by rrPDGF-BB.(2)The results of qRT-PCR:the content ofα-SMA,CollagenⅠ and ATF-2 mRNA decreased significantly with the increase of CPhGs liposome concentration among the different dosage groups(P<0.05),and the inhibition effect of CPhGs liposome 29.45 mg/L was most effective.(3)Western blot analysis:the expression of p-p38 protein decreased in different concentration of CPhGs liposomes with statistical significance(P<0.05).Compared among different dose of CPhGs liposomes,the expression of p-p38 protein decreased significantly with the increase of CPhGs liposomes intervention dose(P<0.05).Conclusion CPhGs liposomes can inhibit the proliferation of HSC induced by rrPDGF-BB,thereby plays the

关 键 词：肝星状细胞 肝纤维化 重组大鼠血小板衍生生长因子-BB 肉苁蓉苯乙醇总苷脂质体 

分 类 号：R332[医药卫生—人体生理学]