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目的:探讨骨髓间质干细胞(MSCs)对活化态肝星状细胞(HSCs)增殖的影响。方法:分别从骨髓和肝脏分离纯化培养大鼠MSCs及HSCs,塑料板传代培养激活HSCs;在半透膜(transwellinsert)上接种MSCs,在6孔塑料培养板上接种HSCs,建立上下双层细胞共培养体系;大鼠正常肝细胞系(BRLs)及HSCs培养分别作为对照。免疫细胞化学检测平滑肌激动蛋白(α-SMA)与结蛋白的表达,IBAS2.5软件分析阳性染色表达量。结果:HSCs与MSCs共培养24h,HSCs表现轻度增殖抑制,随着培养时间延长,HSCs增殖活性受抑制更明显,在48h和72h的抑制率分别达15.7%与30.3%,与BRLs共培养体系比较有显著差异;与MSCs共培养72h,HSCs表达α-SMA量明显低于两个对照组BRLs及HSCs培养体系(50.2%vs90.2%、95.6%,P<0.01);而结蛋白表达的量在3组共培养体系中均无显著差异。结论:MSCs具有分泌细胞因子抑制HSCs增殖活性的潜能,在治疗肝纤维化中可能发挥作用。
Objective: To investigate the effect of bone marrow mesenchymal stem cells (MSCs) on the proliferation of activated hepatic stellate cells (HSCs). METHODS: Rat MSCs and HSCs were isolated and purified from bone marrow and liver, and HSCs were subcultured on plastic plates. MSCs were seeded on transwellinserts and HSCs were seeded on 6-well plastic plates to establish the upper and lower bilayer cells Culture system; rat normal liver cell lines (BRLs) and HSCs culture as control. Immunocytochemistry was used to detect the expression of smooth muscle actin (α-SMA) and desmin. IBAS2.5 software was used to analyze the expression of positive staining. Results HSCs co-cultured with MSCs for 24h, HSCs showed mild proliferation inhibition, with prolonged culture, HSCs proliferation was inhibited more significantly, at 48h and 72h inhibition rates were 15.7% and 30.3%, and BRLs co-culture system (50.2% vs90.2%, 95.6%, P <0.01). However, the expression of desmin in HSCs was significantly lower than that in the two control groups (50.2% vs90.2%, 95.6%, P <0.01) There was no significant difference in the three co-culture systems. Conclusion: MSCs have the potential to inhibit the proliferation of HSCs by secreting cytokines and may play a role in the treatment of hepatic fibrosis.