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中国应用生理学杂志 ›› 2020, Vol. 36 ›› Issue (3): 273-278.doi: 10.12047/j.cjap.5929.2020.060

• 研究论文 • 上一篇    下一篇

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

张倩璐, 旷昕   

  1. 南华大学附属第一医院麻醉科, 湖南 衡阳 421001
  • 收稿日期:2019-08-22 修回日期:2020-04-19 发布日期:2020-09-25
  • 通讯作者: Tel: 18607346516; E-mail: kx6924@126.com
  • 基金资助:
    湖南省自然科学基金省市联合基金项目(2017JJ4049)

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

ZHANG Qian-lu, KUANG Xin   

  1. Department of Anesthesiology, First Hospital Affiliated, University of South China, Hengyang 421001, China
  • Received:2019-08-22 Revised:2020-04-19 Published:2020-09-25

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(5 μmol/L)DMEM培养液培养1 h,用丙泊酚(100 μmol/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。

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

Abstract: 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 (5 μmol/L) for 1 hour, propofol (100 μmol/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 of α-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 of α-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.

Key words: propofol, transforming growth factor-beta 1, hepatic stellate cells, rapamycin target protein in mammals, autophagy

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