钙调神经磷酸酶对钙化血管蛋白激酶G表达的影响

Regulation of expression of cGMP-dependent protein kinase Iα by calcineurin in vascular calcification

目的探讨钙调神经磷酸酶(CaN)对钙化血管蛋白激酶GIα(PKGIα)蛋白及mRNA表达的影响。方法 SD雄性大鼠分为对照组、钙化组及环孢霉素A(CsA)组。采用华法林和维生素D3建立大鼠主动脉钙化模型。9d后HE染色及VonKossa染色观察血管形态及钙沉积改变,应用免疫组化法测定PKGⅠα蛋白及CaNB1蛋白,原位杂交法测定PKGⅠα及CaNAαmRNA,测定CaN和碱性磷酸酶(ALP)活性。结果钙化组PKGIα蛋白和mRNA表达明显减少,与对照组比较,差异有统计学意义(P<0.01),CaNB1蛋白和CaNAαmRNA表达明显增加,与对照组比较,差异有显著性(P<0.05)。结论 CaN能够抑制钙化血管中PKGIα蛋白及mRNA的表达。

Objective To discuss the regulation of expression of cGMP-dependent protein kinase Iα（PKG Iα protein and mRNA）by calcineurin（CaN）in vascular calcification.Methods Rat vascular calcification model was made by subcutaneously injection of warfarin and Vitamin D3.4-weeks-old male SD rats were randomly divided into 3 groups：normal control,calcification and CsA groups.Samples from 6 rats of each group were used for immunohistochemistry to examine the expressions of PKGⅠα protein and CaNB1 protein as well as for in situ hybridization to detect the expressions of PKGⅠα mRNA and CaNAα mRNA.The others were used to detect activities of CaN and alkaline phosphatase（ALP）in rat aorta.Results Expressions of PKGⅠα protein and mRNA in rat aorta from calcification group depressed,and expressions of CaNB1 protein,CaNAα mRNA was elevated significantly.Conclusions CaN can inhibit the expression of PKG Iα protein and mRNA in vascular calcification.