糜酶对肝星状细胞增殖及转分化的影响  

Effects of mast cell chymase on activation, proliferation and transdifferentiation of hepatic stellate cell

作　　者：王洋[1] 尹明实 

机构地区：[1]延边大学附属医院感染症科,延吉130000

出　　处：《免疫学杂志》2016年第1期29-33,共5页Immunological Journal

基　　金：国家自然科学基金(30960372)

摘　　要：目的观察糜酶（chymase）对肝星状细胞（HSC）增殖及转分化的影响,探讨糜酶在肝纤维化中的作用。方法用percoll密度梯度离心法分离、培养大鼠HSC,经系列浓度糜酶的作用后,分别进行如下实验：用噻唑比色法（MTT）检测HSC的增殖;Western blot检测细胞内效应蛋白α-SMA及TGF-β1的表达;用RT-PCR法检测TGF-β1及Ⅰ型胶原（Col-Ⅰ）m RNA的表达。结果肥大细胞（MC）糜酶（1、10、100 ng/ml）明显促进HSC增殖,在观察的范围内（1~100 ng/ml）呈质量浓度依赖性,糜酶质量浓度越高,刺激培养的肝星状细胞增殖的能力越强,最高达对照组（无糜酶刺激）的1.45倍（P〈0.05）;Western blot检测结果显示：糜酶刺激HSC表达α-SMA及TGF-β1明显增强（各实验组与空白对照组比较,P〈0.05）;RT-PCR法检测HSC上TGF-β1及Col-Ⅰm RNA的表达明显增强,此作用呈剂量依赖性（各实验组与空白对照组比较,P〈0.05）。结论肥大细胞糜酶刺激可明显激活HSC,并促进其增殖及转分化为肌成纤维细胞。To observe the effects of mast cell chymase on activation, proliferation and transdifferentiation of hepatic stellate cell（HSC） and to study the role of chymase in development of liver fibrosis, HSC were isolated from rat liver with percoll and treated with chymase, and then divided into groups for further experiments. Proliferation of HSC was measured by means of Methylthiotetrazole（MTT） method; the protein levels of α-SMA and TGF-β1 were detected by Western blotting; and the expression of TGF-β1 m RNA and collagen I m RNA of in vitro cultured HSC were assessed by semi-quantitive reverse transcriptase polymerase chain reaction（RT-PCR）. Data showed that mast cell chymase（1-100 ng/ml） promoted proliferation of HSC and the protein expression of α-SMA and TGF-β1, as well as the m RNA expression of TGF-β1 and collagen I（Col-I） of HSC in a dose-dependent manner（P0.05）. All data indicated that chymase is involved in occurrence and development of liver fibrosis by affecting HSC directly.

关 键 词：肝星状细胞 糜酶 Α-SMA TGF-Β1 胶原 

分 类 号：R575.2[医药卫生—消化系统]