骨髓间充质干细胞诱导肝纤维化大鼠肝星状细胞凋亡和Caspase-3表达  

Investigation of bone marrow mesenchymal stem cells on apoptosis of hepatic fibrosis rats of hepatic stellate cells and the expression of Caspase-3

作　　者：梁梓宇[1] 覃山羽[1] 姜海行[1] 王东旭[1] 苏思标[1] 陈传华[1] 

机构地区：[1]广西医科大学第一附属医院消化内科,南宁530021

出　　处：《重庆医学》2010年第20期2721-2723,共3页Chongqing medicine

基　　金：广西自然科学基金资助项目(0897008);广西自然科学基金资助项目(0640133);广西"新世纪十百千人才工程"基金资助项目(2006206);广西卫生厅青年基金资助项目(2009102)

摘　　要：目的研究大鼠骨髓间充质干细胞(MSCs)诱导肝纤维化大鼠肝星状细胞(HSCs)凋亡及Caspase-3表达。方法分离培养大鼠MSCs。将肝纤维化模型SD大鼠60只平均分为A(鼠尾静脉注射等量的生理盐水)、B(鼠尾静脉注射含MSCs细胞悬液)、C(鼠尾静脉注射经肝细胞生长因子诱导14 d后的MSCs细胞悬液)组。鼠尾静脉输注2×105MSCs细胞悬液,于第1、2、3、4周末取肝组织,经HE、Masson染色观察肝纤维化程度,用TUNEL法检测各组HSCs凋亡,免疫组化检测Desmin,RT-PCR和Western Blot检测凋亡因子Caspase-3。结果 B、C两组肝组织学改善明显,肝纤维化程度减轻,HSCs数量明显减少,Desmin、TUNEL染色阳性的细胞均增多,Caspase-3基因mRNA和蛋白表达增强,且呈时间依赖性,与A组比较差异有统计学意义,且C组较B组明显(P<0.01,P<0.05)。结论 MSCs可在体内诱导HSCs凋亡并上调Caspase-3表达,肝细胞生长因子诱导MSCs抗肝纤维化较单纯MSCs作用更强。Objective To investigate the effect of rat bone marrow mesenchymal stem cells（MSCs） on apoptosis of hepatic fibrosis rats of hepatic stellate cells（HSCs）and the expression of Caspase-3.Methods MSCs were isolated and cultured from bone marrow in rats.Sixty SD hepatic fibrosis rats were divided into 3 groups randomly：A,B and C group.fibrosis model rats were treated with infusion of MSCs suspension or normal saline via tail vein at the beginning of 7th week,after treatment（on weeks of 1,2,3 and 4）,hepatic tissues were taken.The degree of fibrosis was observed under light microscope by hematoxylin and eosin（HE）and Masson staining.The apoptosis of HSCs was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling（TUNEL）.Desmin was detected by immuno-histochemically.The mRNA and protein expressions of Caspase-3 were detected by RT-PCR and Western Blot.Results The hepatic tissue histology of B and C groups obviously improved than A group,the degree of hepatic fibrosis was alleviated,the quantity of HSCs were decreased obviously,the quantity of positive HSCs were increased by Desmin and TUNEL,the mRNA and protein expressions of Caspase-3 were significantly increased at time 1w（P〈0.01,P〈0.05）,and show time dependent.Moreover,we observed that the antifibosis effect of C group was superior to B group.Conclusion MSCs could induce HSCs apotosis in vivo by up-regulating the expression levels of Caspase-3,HGF-induced MSCs was superior to MSCs in the antifibrosis.

关 键 词：骨髓间充质干细胞 肝纤维化大鼠 肝星状细胞 凋亡 CASPASE-3 

分 类 号：R575.2[医药卫生—消化系统]