骨髓间质干细胞培养液上清对肝星状细胞增殖及纤维化的影响

Effect of bone marrow mesenchymal stem cells-conditioned medium on proliferation and fibrogenesis of hepatic stellate cells

背景:利用骨髓间质干细胞治疗肝纤维化的研究取得了一定的效果,但研究其培养液上清治疗肝纤维化的并不多。目的:观察大鼠骨髓间质干细胞培养液上清抑制肝星状细胞增殖及活化的可能性。方法:实验组1,2将骨髓间质干细胞培养液上清加入6孔板内处理肝星状细胞,分别为2mL培养液上清/孔,1mL培养液上清+1mL完全培养基/孔;对照组为肝星状细胞单独培养(2mL完全培养基/孔)。流式细胞仪分析细胞周期并观察抑制效果,RT-PCR检测肝星状细胞内转化生长因子β、金属蛋白酶抑制因子2、胶原蛋白ⅠmRNA的表达情况。结果与结论:加入骨髓间质干细胞培养液上清后,实验组1中的肝星状细胞增殖与对照组相比受到抑制,且抑制率随处理时间的延长而增多(处理72h>处理48h>处理24h,P<0.05);实验组的肝星状细胞内转化生长因子β、金属蛋白酶抑制因子2、胶原蛋白ⅠmRNA的表达较对照组均下调,且下调程度随培养液上清增多而增大(实验组1>实验组2>对照组)。

BACKGROUND：The studies concerning bone marrow mesenchymal stem cells（BMSCs） in treatment of hepatic fibrosis have obtained some outcomes. However,few studies have addressed the conditioned medium for treatment of hepatic fibrosis. OBJECTIVE：To observe how rat BMSCs-conditioned medium to regulate the proliferation and activation of hepatic stellate cells（HSCs） . METHODS：2 mL BMSCs conditioned medium was added in six-well plate to treat the HSCs as experimental group one. 1 mL BMSCs conditioned medium plus 1 mL fresh medium were added to treat HSCs as experimental group two. HSCs were treated with 2 mL fresh medium as the control group. Cell cycle was determined by flow cytometry,and inhibitory effect was observed. The expression of transforming growth factor（TGF） -β,tissue inhibitor of metalloproteinase-2（TIMP-2） ,Collegan I mRNA in HSCs was detected by RT-PCR. RESULTS AND CONCLUSION：After adding BMSCs conditioned medium,in group one,HSC proliferation was significantly inhibited compared with the control group,and the tendency become obvious with the prolongation of time（BMSCs conditioned medium treated 72 h BMSCs conditioned medium treated 48 h BMSCs conditioned medium treated 24 h,P 0.05） . The expressions of TGF-β,TIMP-2,Collegan I mRNA in HSCs were down-regulated in experimental groups one and two compared to the control group,and the tendency was followed by the increase of BMSCs conditioned medium（experimental group one experimental group two control group）.