奥曲肽对肝星状细胞整合素信号通路影响.docVIP

奥曲肽对肝星状细胞整合素信号通路的影响 　　作者：尹凤荣,张晓岚,桑荣霞　　作者单位： 河北省石家庄市第五医院感染一科(尹凤荣);河北医科大学第二医院消化内科(张晓岚);河北省石家庄市第一医院消化内科(桑荣霞) 　　【摘要】目的通过研究奥曲肽对肝星状细胞(HSCs)整合素信号通路的影响，探讨奥曲肽抑制HSCs增殖的机制。方法体外培养HSCs，MTT法测定HSCs增殖，Western blot测定细胞外信号调节激酶(ERK1)蛋白表达。结果干预24 h，Ⅲ～Ⅴ组吸光度(A)值均显著低于Ⅱ组，抑制率分别是18.75%、37.50%和40.63%,差异有统计学意义(P0.05)。Ⅰ组有ERK1蛋白表达，OPN刺激24 h后，ERK1蛋白含量较Ⅰ组高95.62%;Ⅲ～Ⅴ组ERK1蛋白含量则分别低了16.74%、34.30%和65.72%。结论奥曲肽能够通过下调整合素信号通路中的下游信号分子mdash;mdash;ERK1，抑制骨桥蛋白刺激的HSCs增殖。 　　【关键词】 肝纤维化,肝星状细胞,奥曲肽,整合素信号通路 　　【Abstract】 Objective To investigate the action mechanism of octreotide in inhibiting proliferation of hepatic stellate cells (HSCs) through observing the effect of octreotide on the integrin signaling pathway.MethodsThe cells were cultured in vitro,and the proliferation of HSCs was detected by methyl thiazolyl tetrzolium(MTT) assay.The extracelluar signalregulated kinase1 (ERK1) protein expressions in HSCs were analyzed by Western Blot.Results As compared with osteopontin(OPN) group, octreotide at concentrations of 2.5mg/L,5.0mg/L and 10.0mg/L could inhibit the proliferation of HSCs.The inhibition rates were 18.75%,37.50% and 40.63% respectively(P0.05).The expression levels of ERK1 protein in control group were higher,after stimulated by OPN,the protein expression of ERK1 increased by 95.62%.After exposed to octreotide for 24h, ERK1 production decreased by 16.74%,34.30% and 65.72%,respectively for group 2.5mg/L,5.0mg/L and 10.0mg/L.Conclusion The octreotide can inhibit the proliferation of HSCs stimulated by OPN through downregulating ERK1 protein expression, which is the downstream signal molecule of the integrin signaling pathway. 　　【Key words】 hepatic fibrosis;hepatic stellate cell;octreotide;integrin signaling pathway 　　肝纤维化是临床面临的重要问题，其发病机制是一个复杂的级联反应，包括肝星状细胞(hepatic stellate cells，HSCs)活化、增殖、表达各种信号分子及产生大量以胶原为主的细胞外间质(extracellular matrix，ECM)成分[1]。整合素信号通路在活化HSCs增殖过程中发挥重要作用[2]。我们分析奥曲肽对HSCs整合素信号通路的影响，旨在探讨奥曲肽抑制HSCs增殖的机制，寻找新的抗肝纤维化治疗靶点。 　　1 材料与方法 　　1.1 主要试剂与仪器 RMPI1640培养基为美国Gibco公司产