胰岛素和高糖对大鼠肝星状细胞增殖及NF-κB p65蛋白表达的影响

The Effect of Insulin and High Glucose on Proliferation of Hepatic Stellate Cell in Rats and Expression of Protein p65

目的探讨胰岛素和高糖对肝纤维化的影响。方法体外培养大鼠肝星状细胞(hepatic stellate cell,HSC),分为正常对照组、高渗透压对照组、高糖组、(低、中、高)胰岛素组、高糖+(低、中、高)胰岛素组。细胞培养24h和48h,用细胞计数试剂盒-8(CCK-8法)检测HSC的增殖情况,并计算细胞增殖率;细胞培养48h,用免疫印迹法检测HSC NF-κB p65蛋白表达。结果培养24h,各组HSC增殖率与正常对照组相比,差异无统计学意义(P>0.05);培养48h,高、中胰岛素组HSC增殖率明显高于正常对照组和高渗透压对照组(P<0.05);高糖组、(低、中、高)胰岛素组、高糖+(低、中、高)胰岛素组HSC p65蛋白表达量均明显高于正常对照组及高渗透压对照组(P<0.05)。结论胰岛素能诱导体外培养的大鼠HSC增殖,其机制可能与p65蛋白表达增加有关。

Objective To investigate the effect of insulin and high glucose on hepatic fibrosis.Methods Hepatic stellate cells（HSC）of rats were cultured in vitro and divided into five groups including the normal control group,the hyperosmosis control group,the high glucose group,the insulin groups of low,intermediate,and high doses respectively,and the mixed groups of high glucose and insulin of low,intermediate,and high doses respectively.Cell counting kit CCK-8was applied to measure the proliferation of HSC after the cells were cultured for 24 and 48hours respectively and count their proliferation rate.Western blotting was used to detect the HSC protein p65 in each group after the cells were cultured for 48 hours.Results The proliferation rate of HSC in the normal control group after 24 hours was not significantly different from that in the other groups respectively（P〉0.05）.The proliferation rates in the insulin groups of high and intermediate doses were significantly higher after 48 hours than in the normal control group and the hyperosmosis control group（P〈0.05）.The expression levels of HSC p65 protein in the high glucose group,the insulin groups of low,intermediate,and high doses respectively,and the mixed groups of high glucose and insulin of low,intermediate,and high doses respectively were significantly higher than those in the normal control group and the hyperosmosis control group（P〈0.05）.Conclusion Insulin can induce the HSC proliferation in rats in vitro,which may be correlated with the expression increase of Protein p65.