突变型人HGF(tvNK1)对CCl_4诱导的大鼠肝纤维化的影响  

Effects of Human Truncated Variant of Hepatocyte Growth Factor( tvNK1)on Carbon Tetrachloride-induced Liver Fibrosis in Rat

作　　者：王小花[1,2] 李玉婷[1] 刘亚威[1] 桂金秋[1] 周晓杭[1] 袁晓环[2] 初彦辉[2] 刘海峰[2] 

机构地区：[1]牡丹江医学院牡丹江医学院病原微生物与免疫实验室,牡丹江157011 [2]牡丹江医学院黑龙江省抗纤维化重点实验室,牡丹江157011

出　　处：《中国生物工程杂志》2016年第6期18-23,共6页China Biotechnology

基　　金：国家自然科学基金(81573068;81500471);黑龙江省自然科学基金(H2015080);黑龙江省组织损伤与修复重点实验室开放课题(zdsys2013-07;zdsys2013-03)资助项目

摘　　要：目的:探讨突变型人肝细胞生长因子(HGF^(K132E,R134E),tvNK1)对四氯化碳(CCl_4)诱导的SD大鼠肝纤维化的影响。方法:生物发酵大量制备tvNK1,并经腹腔注射于CCl_4诱导的纤维化SD大鼠体内,6周后取肝脏组织,通过real-time PCR和Western blot检测tvNK1对纤维化SD大鼠肝脏中I型胶原蛋白(Collagen type I,Col1A1)、IV型胶原蛋白(Collagen type IV,Col4A1)和α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)在mRNA和蛋白水平表达的影响,并进一步通过HE和Masson染色观察tvNK1对纤维化SD大鼠肝脏形态和肝组织胶原纤维的影响。结果:SDSPAGE检测结果显示,获得纯度≥95%的tvNK1。Real-time PCR和Western blot结果显示,tvNK1降低纤维化大鼠肝脏中Col1A1、Col4A1和α-SMA在mRNA和蛋白水平上的表达。HE和Masson染色结果显示,tvNK1减缓纤维化大鼠肝结构的病理性改变,并降低肝脏中胶原纤维含量。结论:tvNK1能抑制CCl_4诱导的大鼠肝纤维化,为预防肝纤维化和其他器官纤维化疾病提供实验依据。Objective：In order to determine the effects of human truncated variant of hepatocyte growth factor（ tvNK1） on carbon tetrachloride（ CCl4）-induced liver fibrosis in SD rat.Methods：The recombinant tvNK1 was high-level expression in Escherichia coli BL21 by bio-fermentation and rapid purification of tvNK1 protein using Ni^2＋-affinity chromatograph.The SD rat model of CCl4-induced hepatic fibrosis was received intraperitoneal injection of tvNK1 for 6 weeks.The mRNA and protein expression levels of Col1A1、Col4A1 andα-SMA in hepatic tissue of different groups were assayed by real-time PCR and Western blot,respectively.The morphology and the collagen fiber of liver sections in different groups were determined by Hematoxylin and eosin（ HE） and Masson staining,respectively.Results：The high purity tvNK1 was assayed by SDS-PAGE.The recombinant tvNK1 reduced the mRNA and protein expression levels of Col1A1、Col4A1 and α-SMA in CCl4-induced hepatic tissue in Rat.Moreover,injection with tvNK1 restored the morphology and the collagen fiber in CCl4-induced hepatic tissue in Rat.Conclusion：The recombinant tvNK1 protein attenuated progression of hepatic fibrosis in rat and provided a potential treatment for liver fibrosis and other fibrosis diseases.

关 键 词：突变型人肝细胞生长因子 肝纤维化 四氯化碳 胶原纤维 

分 类 号：Q819[生物学—生物工程]