天津医药

天津医药 ›› 2020, Vol. 48 ›› Issue (7): 606-610.doi: 10.11958/20193580

• 细胞与分子生物学 • 上一篇    下一篇

大麻素受体2激动剂AM1241对TGF-β1诱导的HSC-T6增殖、活化及凋亡的影响

龙翠珍1 ，舒远辉1 ，何萍1，2 ，王豫萍1，2△   

  1. 1 贵州医科大学医学检验学院（邮编 550004）；2 贵州医科大学附属医院临床检验中心
  • 收稿日期:2019-12-04 修回日期:2020-04-17 出版日期:2020-07-15 发布日期:2020-07-16
  • 作者简介:龙翠珍（1994），女，硕士在读，主要从事与肝纤维化相关的体内外实验研究

Effects of cannabinoid receptor 2 agonist AM1241 on proliferation, activation and apoptosis of HSC-T6 cells induced by TGF-β1

LONG Cui-zhen1 , SHU Yuan-hui1 , HE Ping1, 2 , WANG Yu-ping1, 2△   

  1. 1 School of Clinical Laboratory Science, Guizhou Medical University, Guiyang 550004, China; 2 Center of Clinical Testing, the Affiliated Hospital of Guizhou Medical University
  • Received:2019-12-04 Revised:2020-04-17 Published:2020-07-15 Online:2020-07-16

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摘要： 目的 探究大麻素受体2激动剂AM1241对转化生长因子（TGF）-β1诱导的大鼠肝星状细胞（HSC-T6）增 殖、活化与凋亡的影响及其作用机制。方法 体外培养HSC-T6，采用CCK-8法检测AM1241对空白对照组（空白培 养基）、阴性对照组（未加药品）、TGF-β1 组（5 μg/L TGF-β1）及 20、40、80、160 μmol/L AM1241 组（5 μg/L TGF-β1+ 20、40、80、160 μmol/L AM1241）HSC-T6增殖的影响，计算半数抑制浓度（IC50）。采用流式细胞术检测阴性对照组、 TGF-β1组、30 μmol/L AM1241组、60 μmol/L AM1241组HSC-T6凋亡情况；采用Western blot检测阴性对照组、TGF- β1 组、27 μmol/L AM1241 组 α-平滑肌肌动蛋白（α-SMA）、碱性成纤维细胞生长因子（bFGF）、凋亡相关蛋白 Bax、 cleaved caspase-3、JNK及磷酸化c-Jun氨基末端激酶（p-JNK）蛋白表达水平。结果 与TGF-β1组比较，AM1241可 抑制HSC-T6增殖能力，且呈剂量依赖性（P＜0.05）。AM1241的IC50为27 μmol/L。30、60 μmol/L AM1241组HSC-T6 凋亡率较TGF-β1组明显上升，且60 μmol/L AM1241组高于30 μmol/L AM1241组（P＜0.05）。27 μmol/L AM1241组 α-SMA 和 bFGF 蛋白表达水平较 TGF-β1 组降低，Bax、cleaved caspase-3、p-JNK 蛋白表达水平较 TGF-β1 组升高 （P＜0.05）。结论 大麻素受体2激动剂AM1241能抑制TGF-β1诱导的HSC-T6的增殖与活化，并促进其凋亡，其作 用机制可能与JNK通路有关。

关键词: 肝硬化；肝星状细胞；受体, 大麻酚, CB2；转化生长因子β1；细胞凋亡；肝纤维化；AM1241

Abstract: Objective To investigate the effects of cannabinoid receptor 2 agonist AM1241 on the proliferation, activation and apoptosis of rat hepatic stellate cells (HSC-T6) induced by TGF-β1 and the related mechanism. Methods HSC-T6 cells were treated with different concentrations of AM1241 (0, 20, 40, 80 and 160 μmol/L) for 24 hours. Cells of each group were given TGF-β1 (5 μg/L) simultaneously. The effect of AM1241 on the proliferation of HSC-T6 cells was detected by CCK-8 assay. HSC-T6 cells were divided into negative control group, TGF- β1 group, 30 μmol/L AM1241 group and 60 μmol/L AM1241 group. Flow cytometry was used to detect the effect of AM1241 on HSC-T6 apoptosis. HSC-T6 cells were divided into negative control group, TGF- β1 group and 30 μmol/L AM1241 group (according to CCK-8 assay). IC50 was calculated. The protein levels of α-smooth muscle actin (α-SMA), basic fibroblast growth factor (bFGF), apoptosis-related protein Bax, cleaved caspase-3 and phosphorylated c-Jun amino terminal kinase (p-JNK) were detected by Western blot assay. Results Compared with TGF-β1 group, AM1241 showed an inhibitory effect on the proliferation of HSC-T6 cells in a dose-dependent manner (P＜0.05). When IC50 of AM1241 was 27 μmol/L, the apoptosis rates of HSC-T6 were significantly higher in AM1241 groups (30 and 60 μmol/L) than those in TGF - β1 group, and it was significantly higher in 60 μmol/L AM1241 group than that of 30 μmol/L AM1241 group (P＜0.05). The protein levels of α-SMA and bFGF were significantly lower in 27 μmol/L AM1241 group than those in the TGF-β1 group (P＜0.05). The protein levels of Bax, cleaved caspase-3 and p-JNK were significantly upregulated in AM1241 group than those of the TGF-β1 group (P＜0.05). Conclusion The cannabinoid receptor 2 agonist AM1241 can inhibit the proliferation and activation of HSC-T6 cells induced by TGF-β1 and promote the apoptosis of HSC-T6 cells, which may be related to the JNK pathway.

Key words: liver cirrhosis, hepatic stellate cells, receptor, cannabinoid, CB2, transforming growth factor beta1; apoptosis, liver fibrosis, AM1241