重组人松弛素-2对肝星状细胞增殖、Ⅰ型胶原合成及TGF-β1与CTGF mRNA表达的影响

摘要

目的：在细胞水平研究松弛素的抗肝纤维化作用,并初步探讨其分子机制,为肝纤维化的治疗提供实验依据。方法体外培养大鼠肝星状细胞(HSC-T6),分别用20、50、100 ng/mL重组人松弛素-2(recombinant human relaxin-2, RLX-2)干预,采用噻唑蓝(MTT)比色实验检测 RLX-2对 HSC-T6增殖的影响,双抗体夹心酶联免疫吸附法(ELISA)检测对照组及RLX-2作用48 h后的细胞培养基上清液Ⅰ型胶原水平,利用逆转录酶-聚合酶链反应(RT-PCR)技术检测药物干预48 h的结缔组织生长因子(CTGF)、转化生长因子-β1(TGF-β1)mRNA 表达情况。结果 RLX-2对 HSC的增殖具有抑制作用,可降低 HSCⅠ型胶原水平,RLX-2抑制 HSC的CTGF mRNA表达,对 HSC的TGF-β1 mRNA表达无明显影响。结论 RLX-2可能通过抑制体外培养大鼠 HSC 细胞的增殖、Ⅰ型胶原及 CTGF mRNA 的表达而发挥抗大鼠肝纤维化作用。%ABSTRACT:Objective To validate that relaxin can resist hepatic fibrosis at the cellular level and explore its molecular mechanism in order to provide experimental basis for the treatment of liver cirrhosis.Methods Cultured HSC-T6s were treated with different concentrations (20,50 and 100 ng/mL)of recombinant human relaxin-2 (RLX-2).The proliferation of HSC-T6 was measured by MTT colorimetric assay.The content of type Ⅰcollagen in the cell culture supernatant of each group was detected by ELISA at 48 h of drug intervention;RT-PCR was used to detect the mRNA expressions of CTGF and TGF-β1 in HSC-T6 at 48 h of drug intervention.Results RLX-2 inhibited the proliferation of HSC and reduced type Ⅰ collagen content of HSC cells.It also inhibited the CTGF mRNA expression of HSC,but did not have a significant effect on the expression of TGF-β1 mRNA. Conclusion In the experiment of culturing HSC-T6 in vitro,RLX-2 may play a role in rat liver fibrosis by inhibiting cell proliferation and type Ⅰ collagen and CTGF mRNA expressions.