白细胞介素6体外诱导骨髓间充质干细胞向肝细胞的分化

Interleukin 6 induces the differentiation of bone marrow mesenchymal stem cells into hepatocytes in vitro

背景：骨髓间充质干细胞是肝细胞的重要肝外来源,在多种细胞因子和生长因子诱导下可分化为肝细胞,从而参与肝功能的修复和重建。目的：观察白细胞介素 6 是否可诱导骨髓间充质干细胞向肝细胞定向分化。方法：采用贴壁法分离培养昆明种雄性小鼠骨髓间充质干细胞,在无血清肝细胞培养液 HEPATOZYME-SFM 中分别加入2.5,5,10,20 μg/L 白细胞介素 6 诱导其向肝样细胞分化;诱导培养 0,7,14,21,28 d 时取出细胞爬片,细胞免疫组织化学法检测细胞角蛋白 18、甲胎蛋白表达情况,过碘酸-Schiff 糖原染色检测细胞功能,ELISA 和硝酸还原酶法分别检测培养液中白蛋白和一氧化氮水平。结果与结论：当白细胞介素 6 质量浓度在 2.5~10 μg/L 范围内,呈浓度依赖性和时间依赖性增加细胞角蛋白 18、糖原表达率及细胞培养液中白蛋白水平;甲胎蛋白表达为先升后降直至 21d 停止。当白细胞介素 6 质量浓度增加到 20 μg/L 时上述诱导作用均不及质量浓度 10 μg/L 组。只有 10 μg/L 白细胞介素 6 组诱导到 28 d 时细胞培养液中测得微量一氧化氮;提示白细胞介素 6 可诱导骨髓间充质干细胞向肝系细胞分化,且在 10 μg/L 作用最强,质量浓度再增高时诱导作用反而减弱。

BACKGROUND：Bone marrow mesenchymal stem cells（BMSCs） are an important source of hepatocytes.BMSCs can differentiate into hepatocytes under the induction of multiple cytokines and growth factors,and participate in repair and reconstruction of liver function.OBJECTIVE：To explore the effect of interleukin 6 on the differentiation of BMSCs into hepatocytes.METHODS：BMSCs were isolated from Kunming male mice and cultured by attachment method and treated with interleukin 6（2.5,5,10,20 μg/L） in HEPATOZYME-SFM nutrient solution to induce the BMSCs differentiation into hepatocytes.On days 0,7,14,21,28 after induction,the expressions of cytokeratin 18,alpha-fetoprotein and glycogen were identified by immunohistochemical method and periodic acid-schiff.The contents of albumin and nitric oxide in the liquid supernatant were identified by ELISA and nitrate reductase method,respectively.RESULTS AND CONCLUSION：Interleukin 6 resulted in dose-dependent（2.5,5,10 μg/L） and time-dependent increase in the expressions of cytokeratin 18 and glycogen and the content of albumin.The expression of alpha-fetoprotein increased then decreased to disappearance on day 21.The above effects of interleukin 6（20 μg/L） was weaker than that of interleukin 6（10 μg/L）.The content of small amount nitric oxide was detected when interleukin 6 was 10 μg/L on day 28.These demonstrated that interleukin 6 is able to induce the differentiation of BMSCs into hepatocyte-like cells,which shows strongest effects at 10 μg/L.Increased mass concentration showed decreased induction effects.