RNA干涉干扰素调节因子-1和CD74可抑制因高糖所致自发高血压大鼠大血管平滑肌细胞的过度增殖

目的:探讨特异性iRNA下调干扰素调节因子-1(IRF-1)及CD74表达对高糖所致自发高血压大鼠源大血管平滑肌细胞的过度增殖的影响.方法:合成特异性IRF-1及CD74的RNA干扰片段,并将其插入载体pGSsi-U6/Neo/GFP以构建重组载体.借助Lipofectmine2000,将重组载体转染自发高血压大鼠源大血管平滑肌细胞,然后将细胞置于高糖环境培养.细胞计数、MTT法及PCNA表达以检测平滑肌细胞的增殖能力.结果:携带特异性干扰IRF-1和CD74片段的重组质粒,经Lipofectmine2000转染后,分别见88.9%和80.5%的平滑肌细胞显示绿色荧光.与对照组相比,RNA干扰后,CD74和IRF-1的表达降至9.2%和4.2%.CD74和IRF-1表达的缺失导致细胞活力和PCNA表达的显著下调.MTT法的结果显示,IRF-1干扰引起平滑肌细胞OD的限制下降,但类似的结果在CD74干扰组中未见.结论:上述结果提示IRF-1和CD74基因与糖尿病大血管病变相关,或可作为该疾病的治疗靶点.

Knockdowning interferon regulatory factor-1 and CD74 suppress high glucose induced access proliferation of macrovascular smooth muscle cells derived from spontaneously hypertensive rats

Objective:The goal of this study was to explore the influence of specific RNA interference targeting CD 74 and IRF-1 on accelerated proliferation of macrovascular smooth muscle cells induced by high glucose media. Methods:The specific small RNA interfering segments targeting CD74 and IRF-1 were synthesized and inserted into pGSsi-U6/Neo/GFP. Mediated by transfection reagent LipofectMINE 2000, the recombinant vector containing specific small interfering RNA segment were transfected into macrovascular smooth muscle cells and cultured in high glucose. Proliferative capability of VSMCs was examined by cell counting, MTT assay and PCNA expression. Results:Mediated by Lipofectmine 2000, transfection with recombinant vectors containing specific small interfering RNA segment targeting CD74 and IRF-1 caused 80.5%and 88.9%of macrovascular smooth muscle cells exhibiting positive to GFP respectively. Compared with those of control, only 9.2%of CD74 and 4.2%of IRF-1 were detected in mRNA level. Lack of IRF-1 and CD74 functions resulted in a significant decline in survivals and PCNA expression of VSMCs cultured in high glucose media. A significant decrease in optic density was detected in IRF-1 knockdowning cells, but not in CD74 silence group. Conclusion:Present data suggest that IRF-1 and CD74 genes are linked to VSMCs proliferation in diabetic macroangiopathy, and have a potential as a promising target to reduce the macroangiopathic formation of diabetes.