丙泊酚对转化生长因子-β1诱导肝星状细胞激活的影响及其机制  

Effects of propofol on TGF-β1 induced activation of hepatic stellate cells and its mechanism

作　　者：张倩璐[1] 旷昕[1] ZHANG Qian-lu;KUANG Xin(Department of Anesthesiology,First Hospital Affiliated,University of South China,Hengyang 421001,China)

机构地区：[1]南华大学附属第一医院麻醉科,湖南衡阳421001

出　　处：《中国应用生理学杂志》2020年第3期273-278,共6页Chinese Journal of Applied Physiology

基　　金：湖南省自然科学基金省市联合基金项目(2017JJ4049)。

摘　　要：目的:观察丙泊酚对转化生长因子-β1(TGF-β1)诱导的肝星状细胞系HSC2-T6细胞激活的影响并探讨其可能的作用机制。方法:实验分为对照组、TGF-β1组、丙泊酚组、TGF-β1+丙泊酚组、雷帕霉素组、TGF-β1+丙泊酚+雷帕霉素组。先用含雷帕霉素(5μmol/L)DMEM培养液培养1 h,用丙泊酚(100μmol/L)处理1 h,然后再加入TGF-β1(5 ng/ml)继续共同培养24 h。细胞的增殖水平通过MTT法测量,细胞培养液上清中透明质酸(HA)、IV型胶原(IV-C)和层粘连蛋白(LN)的浓度采用ELISA法测量,细胞的超微结构采用透射电镜观测,α-平滑肌肌动蛋白(α-SMA)、哺乳动物雷帕霉素靶蛋白(mTOR)、磷酸化mTOR(p-mTOR)及自噬相关基因Beclin 1、微管相关蛋白1轻链3(LC3)和p62的表达通过Western blot测量。结果:与对照组比较,TGF-β1组细胞增殖、α-SMA蛋白的表达、培养液上清中HA、IV-C和LN的浓度、自噬体数量、Beclin-1和LC3-Ⅱ的蛋白表达及LC3-Ⅱ/LC3-Ⅰ比值显著性增加(P均<0.05),p-mTOR蛋白的表达和p-mTOR/mTOR比值及p62的蛋白表达显著性降低(P均<0.05)。与TGF-β1组比较,丙泊酚+TGF-β1组细胞增殖、α-SMA蛋白的表达、培养液上清中HA、IV-C和LN的浓度、自噬体数量、Beclin-1和LC3-Ⅱ的蛋白表达及LC3-Ⅱ/LC3-Ⅰ比值均显著性降低(P均<0.05),p-mTOR蛋白表达和p-mTOR/TOR比值及p62的蛋白表达均显著性增加(P均<0.05)。mTOR抑制剂雷帕霉素部分逆转丙泊酚的作用。结论:丙泊酚抑制TGFβ1诱导的肝星状细胞激活,其机制涉及mTOR-自噬途径。Objective:To observe the effects of propofol on the activation of hepatic stellate cell line HSC2-T6 induced by transforming growth factor-beta 1(TGF-β1)and explore its possible mechanism.Methods:The cells were divided into control group,TGF-β1 group,propofol group,TGF-β1+propofol group,rapamycin group,TGF-β1+propofol+rapamycin group.Cells were treated with rapamycin(5μmol/L)for 1 hour,propofol(100μmol/L)for 1 hour,then TGF-β1(5 ng/ml)was added to co-culture for 24 hours.Cell proliferation was measured by MTT assay.The concentrations of hyaluronic acid(HA),collagen IV(IV-C)and laminin(LN)in the supernatant of cell culture medium were measured by ELISA.The ultrastructure of cells was observed by transmission electron microscopy.The expressions of alpha-smooth muscle actin(α-SMA),mammalian rapamycin target protein(mTOR),phosphorylated mTOR(p-mTOR)and the autophagy related gene Beclin 1,LC3 and p62 were measured by Western blot.Results:Compared with control group,cell proliferation,the expression ofα-SMA,the concentrations of HA,IV-C and LN in culture supernatant,the number of autophages,the expressions of Beclin-1 and LC3-II,the ratio of LC3-II/LC3-I in HSC2-T6 cells were increased significantly,while the expression of p-mTOR,the ratio of p-mTOR/mTOR and the expression of p62 protein were decreased significantly in TGF-β1 group(All P<0.05).Compared with TGF-β1 group,cell proliferation,the expression ofα-SMA,the concentrations of HA,IV-C and LN in culture supernatant,the number of autophages,the expressions of Beclin-1 and LC3-II,the ratio of LC3-II/LC3-I in HSC2-T6 cells in TGF-β1 group were decreased significantly,and the expression of p-mTOR,the ratio of p-mTOR/mTOR and expression of p62 protein were increased significantly in TGF-β1+propofol group(All P<0.05).Conclusion:Propofol inhibits the activation of hepatic stellate cells induced by TGF-beta 1,and its mechanism involves the mTOR-autophagy pathway.

关 键 词：丙泊酚 转化生长因子-Β1 肝星状细胞 哺乳动物雷帕霉素靶蛋白 自噬 

分 类 号：R965[医药卫生—药理学]