脂肪间质干细胞抑制肝星状细胞增殖活化及肝脏纤维化  

Adipose-derived mesenchymal stem cells inhibit the proliferation and activation of hepatic stellate cell in vitro and inhibit rat liver fibrogenesis in vivo

作　　者：俞富祥[1] 苏龙丰[1] 季世强[1] 张启瑜[1] 

机构地区：[1]浙江省温州医学院附属第一医院肝胆胰外科,325000

出　　处：《中华普通外科杂志》2011年第12期1027-1030,共4页Chinese Journal of General Surgery

摘　　要：目的 探讨脂肪间质干细胞（adipose tissue-derived mesenchymal stem cells,ADSCs）对活化态肝星状细胞（hepatic stellate cells,HSCs）增殖、活化的影响及其对活体肝硬化模型的治疗作用.方法 分别分离纯化培养HSCs及ADSCs,通过半透膜（transwell insert）建立HSCs与ADSCs的双层培养体系,大鼠正常肝细胞系（（buffalo rat liver cells,BRLs）培养作为对照.通过CCK-8比色法检测ADSCs对HSCs增殖的影响；Western blot检测HSCs的α-肌动蛋白（α-SMA）表达；建立鼠肝硬化模型,经门静脉输注ADSCs或BRLs,检测肝组织肝硬化；通过检测培养液中细胞因子的含量探讨可能的作用机制.结果 成功分离获得原代HSCs与ADSCs,活化的HSCs与ADSCs共培养72 h,与对照组相比,ADSCs明显抑制HSCs的活化（各组灰度值分别为1.4±0.2,152±14,258±18,F=283.348,P＜0.05）与增殖（各组吸光度A值分别为2.172±0.107,1.424±0.013,1.209±0.117,F=90.605,P ＜0.05）,活体移植显示ADSCs对肝硬化具有明显抑制作用（分别F=77.234、65.164、58.309,均P ＜0.05）.培养液细胞因子检测显示ADSCs比BRLs分泌更多的肝细胞生长因子（hepatocyte growth factor,HGF）（F=0.005,P＜0.05）,分泌较少的转化生长因子（transforming growth factor-β,TGF-β1）（F=1.767,P＜0.01）及神经生长因子（nerve growth factor,NGF）（F=2.301,P＜0.05）. 结论 ADSCs具有分泌细胞因子抑制HSCs活化增殖的潜能,对活体肝硬化进程具有一定的抑制作用.Objective To investigate the effect of adipose derived stem cells （ADSCs） on hepatic stellate cells （HSCs） in vitro and on liver fibrosis in vivo.Methods ADSCs and HSCs were isolated from adipose tissue and liver respectively in SD rats.The coculture system was set up by transwell insert.The 5th passage HSCs were cultured on the 6-well plastic plate,and ADSCs or BRLs seeded on the transwell insert.The proliferation of HSCs was tested by CCK-8 test kit.Smoothmuscle α-actin （α-SMA） expression of HSCs were tested by Western blot.Rat models of liver fibrosis was established.Rats in ADSCs treatment group were infused with ADSCs and those in control group were infused with Buffalo rat liver cells （BRLs）.Liver sections were studied by immunocytochemistry.Liver hydroxyproline （Hyp） content,serum laminin （LN）and hyaluronic acid （HA） were tested,the cytokines in the culture medium were assayed.Results HSCs and ADSCs were isolated successfully.After coculture for 72 h,compared with the control group,the proliferation and activation of HSCs was inhibited by ADSCs（ absorbance of each group were 2.172 ±0.107,1.424 ± 0.013,1.209 ± 0.117,F =90.605,P 〈 0.05 ; Gray-scale values of each group were 1.4 ± 0.2,152 ± 14,258 ± 18,F =283.348,P 〈 0.05 ）,ADSCs infusion inhibits liver fibrosis in model rats （ F =77.234,65.164,58.309,all P 〈 0.05 ）.More hepatocyte growth factor（HGF） and less transforming growth factor-β1 （TGF-β1） （F=1.767,P〈0.05）and nerve growth factor （NGF） （F=2.301,P〈0.05） were secreted by ADSCs than by BRLs.Conclusions ADSCs inhibit the proliferation and activation of hepatic stellate cells.Treatment with ADSCs decreases collagen deposition in the liver and inhibits liver fibrosis.

关 键 词：肝硬化 实验性 间质干细胞 增殖 肝星状细胞 

分 类 号：R575.2[医药卫生—消化系统]