鳖甲软肝健脾方对肝星状细胞活化及细胞外基质代谢的影响  

Effects of Biejia Ruangan Jianpi Formula on activation of hepatic stellate cells and extracellular matrix metabolism

作　　者：刘洋[1] 李彤[1] 文彬 何春雨 钟晓丹 孙海涛[1] 贺松其[1] LIU Yang;LI Tong;WEN Bin;HE Chun-yu;ZHONG Xiao-dan;SUN Hai-tao;HE Song-qi(Southern Medical University,Guangzhou 510515,China;Air Force Hospital of Southern Theater Command of PLA,Guangzhou 510602,China)

机构地区：[1]南方医科大学,广州510515 [2]解放军南部战区空军医院,广州510602

出　　处：《中华中医药杂志》2022年第10期5643-5647,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金面上项目(No.81774037)。

摘　　要：目的:通过研究鳖甲软肝健脾方对肝星状细胞(HSC)活化及Wnt/β-连环蛋白(β-catenin)信号通路相关信号分子蛋白表达水平的影响,探讨鳖甲软肝健脾方抗肝纤维化的作用机制。方法:运用血清药理学,以鳖甲软肝健脾方,鳖甲软肝健脾方拆方扶正组、拆方祛邪组,鳖甲煎丸含药血清干预HSC-T6细胞,24 h后,采用CCK8、凋亡荧光、流式细胞术、Western Blot检测鳖甲软肝健脾方对HSC活化、细胞外基质代谢[α-平滑肌肌动蛋白(α-SMA)、转化生长因子β1(TGF-β1)、组织金属蛋白酶抑制物-1(TIMP-1)、基质金属蛋白酶(MMP)-2、MMP-9]及Wnt/β-catenin信号通路[β-catenin、糖原合成酶激酶3β(GSK-3β)、p-GSK-3β、p-β-catenin]中相关信号分子蛋白表达水平的影响。结果:与阴性对照组比较,鳖甲软肝健脾方可抑制HSC-T6细胞增殖并促进其凋亡,同时,可显著抑制HSC-T6细胞中α-SMA、TGF-β1、TIMP-1、p-GSK-3β蛋白表达(P<0.05,P<0.01),促进MMP-2、MMP-9、p-β-catenin蛋白表达(P<0.05)。结论:鳖甲软肝健脾方可抑制HSC活化,并能促进细胞外基质降解,其机制可能与抑制Wnt/β-catenin信号通路激活有关。Objective: To study the effects of Biejia Ruangan Jianpi Formula on the activation of hepatic stellate cells and the expression of signal molecules related to Wnt/β-catenin signal pathway, and to explore the mechanism of Biejia Ruangan Jianpi Formula on anti-hepatic fibrosis. Methods: Using serum pharmacology, the drug-containing serum of Biejia Ruangan Jianpi Formula,Biejia Ruangan Jianpi prescription Fuzheng Group, disassembled prescription Quxie Group and Biejiajian Pill were used to interfere with HSC-T6 cells. After 24 hours, CCK8, apoptosis fluorescence, flow cytometry and Western Blot were used to detect the effects of Biejia Ruangan Jianpi Formula on the activation of hepatic stellate cells, extracellular matrix metabolism(α-SMA, TGF-β1,TIMP-1, MMP-2, MMP-9) and the expression of related signal molecules in Wnt/β-catenin signal pathway(β-catenin, GSK-3β,p-GSK-3β, p-β-catenin). Results: Compared with the negative control group, Biejia Ruangan Jianpi Formula could inhibit the proliferation and promote the apoptosis of HSC-T6 cells. At the same time, it could significantly inhibit the expression of α-SMA,TGF-β1, TIMP-1 and p-GSK-3β protein in HSC-T6 cells(P<0.05, P<0.01), and promote the expression of MMP-2, MMP-9 and p-β-catenin protein(P<0.05). Conclusion: Biejia Ruangan Jianpi Formula can inhibit the activation of hepatic stellate cells and promote the degradation of extracellular matrix. The mechanism may be related to its inhibition of the activation of Wnt/β-catenin signal pathway in HSC-T6 cells.

关 键 词：鳖甲软肝健脾方 肝星状细胞 细胞凋亡 细胞外基质 WNT/Β-CATENIN信号通路 

分 类 号：R285[医药卫生—中药学]