甲基莲心碱对转化生长因子-β1诱导的肝星状细胞增殖及gremlin表达的影响  

The impact of neferine on the proliferation of transforming growth factor-pl-induced hepatic stellate cells and the expression of gremlin

作　　者：王雅琴[1] 陈永平[1] 刘真真[1] 宋梅[1] 陈少隆[1] 林镯[1] 沈宇娟[1] 陈黎黎[1] 戴春蕾[1] 

机构地区：[1]温州医学院附属第一医院感染内科,325000

出　　处：《中华传染病杂志》2013年第3期144-149,共6页Chinese Journal of Infectious Diseases

摘　　要：目的观察甲基莲心碱对转化生长因子（TGF）-β1诱导的肝星状细胞（HSC）增殖及gremlin、TGF-β1、α-平滑肌动蛋白（α-SMA）表达的影响。方法体外培养HSC-T6细胞，分成正常对照组、TGF-131干预组、TGF一81＋甲基莲心碱分别为2．0、4．0、8．0mmol／L联合干预组，细胞计数试剂盒（cCK）一8检测细胞增殖抑制率，半定量RT—PCR．Western印迹法分别检测HSC—T6细胞gremlin、TGF-β1、α-SMAmRNA及蛋白表达。组间差异采用单因素方差分析，方差齐性者两两比较采用LSD法，方差不齐者采用DunnettT3检验，相关性采用Pearson直线相关分析。结果与正常对照组相比，TGF-β1对HSC—T6细胞增殖无影响（P〉0．05），但能上调gremlin、TGF-β1、α-SMAmRN『A及蛋白的表达，差异有统计学意义（P〈0．05）。与TGF-β1干预组相比，甲基莲心碱能抑制HSC—T6细胞增殖，呈时间、剂量依赖性，以TGF-β1＋甲基莲心碱8mmol／L联合作用72h对HSC—T6细胞抑制最显著（F=2817．056，P〈O．01），抑制率为（69．08±1．05）％。甲基莲心碱可不同程度抑制gremlinmRNA及蛋白的表达，TGF-131＋8mmol／L甲基莲心碱联合干预组表达量最低，为0．2483±0．0084、0．9307±0．0224。与TG-β1干预组比较，gremlin、TGF-β1、α-SMAmRNA表达量随甲基莲心碱浓度的增加呈递减趋势（F=16．800、209．246、70．486，均P〈0．05），蛋白表达量也呈递减趋势（F一56．588、12．550、32．141，均P〈0．05）。gremlin、TGF-β1、a—SMA的表达呈两两正相关（P〈O．05）。结论甲基莲心碱能有效作用于HSC，gremlin可能参与肝纤维化的发生发展，有望成为肝纤维化治疗新靶点。[Abstract] Objective To observe the impact of neferine （Nef） on the proliferation of transforming growth factor-~l （TGF-β）-induced hepatic stellate cells （HSC） and the expression of gremlin and TGF-[31, α-smooth muscle actin （α-SMA）. Methods HSC-T6 ceils were cultured in vitro and divided into five groups：control group, TGF-β1 group, TGF-J31α-Nef 2.0 mmol/L group,TGF-β1 ＋ Nef 4.0 mmol/L group and TGF-β1 α-Nef 8.0 mmol/L group. Inhibition of proliferation of HSC-T6 cells was measured by cell counting kit-8. Semi-quantitative reverse transcription-polymerase chain reaction （RT-PCR） and Western blot assay were used to evaluate the mRNA and protein expressions of gremlin, TGF-β1 and α-SMA, respectively. The differences among groups were compared by one- way analysis of variance. Comparison between two groups were analyzed by LSD-t test and Dunnett T3 test. Correlation was tested by Pearson linear correlation analysis. Results TGF-β1 had no effecton the proliferation of HSC-T6 cells when compared with the control group （P〈0.05）, but it could up-regulate the mRNA and protein expression of gremlin, TGF-β1 and α-SMA （P〈0.05）. The proliferation of HSC-T6 cells were inhibited by Nef in a time and dose dependent way, when compared with the TGF-I]I group. The most significant inhibition was observed in the TGF-I]!＋Nef 8 mmol/L group at 72 hours （F= 2817. 056, P〈0. 01）, and the inhibition rate was （69.08 zk 1.05）0/60. The expression of gremlin mRNA and protein could be inhibited by Nef in varying degrees, to be the lowest in the TGF-β1 ＋ Nef 8mmol/L group （0.2483~0.（1084 and 0.9307~0.0224, respectively）. Compared with the TGF-β1 group, both the mRNA and protein expressions of gremlin, TGF-Ol and α-SMA were gradually decreased with the increasing concentrations of Nef （F= 16. 800, 209. 246 and 70. 486, respectively~ all P〈 0. 05 F = 56. 588, 12. 550 and 32. 141, respectively all P〈 0.05）, respectively. The expressions of gremlin, TGF-β1 and α-SM

关 键 词：生物碱类 骨形态发生蛋白质类 转化生长因子-Β1 肝硬化 星形细胞 

分 类 号：R285.5[医药卫生—中药学]