罗格列酮对高糖诱导大鼠血管平滑肌细胞炎症和增殖的影响

Effects of rosiglitazone on inflammation and proliferation of rat vascular smooth muscle cells induced by high glucose

目的探讨罗格列酮(RSG)对高浓度葡萄糖孵育下的血管平滑肌细胞(VSMCs)炎症和凋亡的影响及其可能的分子机制。方法以不同浓度的葡萄糖和罗格列酮单独或联合孵育大鼠胸主动脉平滑肌细胞,ELISA方法检测培养基中单核细胞趋化蛋白-1(MCP-1)的水平;采用流式细胞术检测VSMCs细胞凋亡率及Bcl-xl、Bcl-2蛋白表达;Western印迹检测胞浆中VSMCsBcl-xl蛋白表达及NF-κBp65和IκBα的表达。结果高葡萄糖浓度(11.2、22.5mmol/L)培养可明显增加上清中MCP-1的浓度,促进VSMCs增殖,抑制其凋亡,上调Bcl-xl、Bcl-2蛋白表达,同时使NF-κBp65表达增加,IκBα表达下降;30及100μmol/LRSG以浓度依赖形式减少VSMCs对MCP-1的分泌,抑制高葡萄糖培养下(11.2、22.5mmol/L)VSMCsBcl-xl、Bcl-2表达,促进其凋亡;下调NF-κBp65表达,促进IκBα表达。RSG拮抗剂GW9662(10μmol/L)预处理可部分拮抗RSG的作用。结论 RSG可能通过对NF-κB通路的调控,减少炎症因子MCP-1分泌,下调Bcl-xl、Bcl-2表达,从而抑制高糖葡萄糖培养下的VSMCs炎症反应并促进其凋亡,从而在2型糖尿病大血管病变的防治中发挥重要作用。

Objective To investigate the effects of rosiglitazone（RSG）on inflammation and apoptosis in rat vascular smooth muscle cells（VSMCs）induced by high glucose administration.Methods Rat VSMCs were incubated with glucose in different concentrations in the presence or absence of RSG or/and RSG antagonist GW9662.The concentration of MCP-1 in the supernatant was measured with ELISA.The apoptotic rates and the protein expressions of Bcl-xl and Bcl-2 were examined by flow cytometry.The Bcl-xl protein expression and NF-κB p65,IκB protein expressions were also evaluated by Western blotting.Results High glucose（11.2,22.5 mmol/L）significantly increased MCP-1 secretion,inhibited apoptosis,upregulated Bcl-xl and Bcl-2 expression in rat VSMCs.Meanwhile,NF-κBp65 expression was increased and IκB protein expression decreased.RSG at 30,100 μmol/L,in a concentration-dependent manner,significantly inhibited MCP-1 secretion,promoted the cells to apoptosis and decreased expression of Bcl-2,Bcl-xl protein induced by high glucose.Also NF-κBp65 expression was downregulated accompanied with upregulation of IκB.The effects of RSG were partly attenuated after pretreated with RSG antagonist GW9662（10 μmol/L）.Conclusions Under high glucose concentration,RSG can reduce the proinflammatory mediator MCP-1 secretion,down-regulate the protein expressions of Bcl-xl and Bcl-2 through modulating the NF-κB pathway,subsequently inhibit the inflammatory and proliferative responses in VSMCs,which suggests that RSG may play a protective role against diabetic macroangiopathy.