脂肪间质干细胞对肝星状细胞活化、增殖、凋亡相关作用的研究

Regulation Effect of Adipose-derived Mesenchymal Stem Cells on Hepatic Stellate Cells in Vitro

目的探讨脂肪间质干细胞(ADSCs)对活化态肝星状细胞(HSCs)增殖、活化凋亡的影响及其探讨产生作用的原因。方法从肝脏及腹腔脂肪分别分离纯化培养HSCs及ADSCs,通过半透膜(transwell insert)在6孔塑料培养板上建立HSCs与ADSCs的双层培养体系,大鼠正常肝细胞系(BRLs)及HSCs培养分别作为对照。通过CCK-8比色法检测ADSCs对HSCs增殖的影响;Western blotting检测HSCsα-肌动蛋白(α-SMA)的表达,了解HSCs的活化状态;流式细胞仪检测ADSCs对HSCs凋亡的影响。最后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 HSCs与ADSCs共培养72h,ADSCs明显抑制HSCs的活化与增殖,促进HSCs的凋亡,培养液细胞因子检测显示ADSCs比BRLs分泌更多的HGF,而较少分泌TGF。结论 ADSCs具有分泌细胞因子抑制HSCs活化增殖,促进凋亡的潜能。ADSCs的治疗作用可能与其分泌的细胞因子有关。

Objective To investigate the regulation effect of adipose derived stem ceils （ADSCs） on hepatic stellate cells （HSCs） in vitro. Methods ADSCs and HSCs were isolated from adipose tissue and liver respectively in SD rats. The coculture system was set up by transwell insert with pore size 0.4μm and 6 - well plates. The 5th passage HSCs were cultured on the 6 - well plastic plate, An ADSCs or buffalo rat liver cells （BRLs） or HSCs seeded on the transwell insert. The proliferation of HSCs was tested byCCK - 8 test kit. Smoothmuscleoα--actin （α-SMA） expression of HSCs were tested by immunocytochemistry. The apoptosis of HSCs was tested by flow cytometer. The cytokines in the culture solution were tested by ELISA kit. Results After coculture for 72h, the proliferation and activation of HSCs was inhibited by ADSCs and the apoptosis of HSCs was promoted by ADSCs. More HGF was secreted by ADSCs than by BRLs, but less TGF was secreted by ADSCs than by BRLs. Conclusion ADSCs can inhibit the proliferation and activation of hepatic stellate cells. One of the reasons for inhibition of liver fibrogenesis by ADSCs maybe that ADSCs secret some cytokines.