高糖通过下调miR-23b促进血管平滑肌细胞的增殖和迁移

High Glucose Induced Vascular Smooth Muscle Cell Proliferation and Migration Through Suppressing mi R-23b

目的高糖诱导血管平滑肌的增殖和迁移是糖尿病引起动脉粥样硬化的关键病理环节。小分子RNA在糖尿病血管并发症中的作用日益受到重视,本项目将对微小RNA-23b(miR-23b)在高糖诱导血管平滑肌的增殖和迁移中的作用进行研究。方法首先,将血管平滑肌细胞株(VSMCs)与高糖(HG,40 m M)共同孵育24 h,q PCR检测高糖对VSMCs增殖及miR-23b的表达的影响。为了进一步证实miR-23b在高糖诱导VSMC增殖及迁移中的作用,转染过表达miR-23b载体及对照载体,构建过表达VSMCs(VSMC^(miR-23b))及对照细胞(VSMCNC),将HG分别与VSMC^(miR-23b)及VSMCNC孵育24 h,CCK-8检测VSMCs的增殖,Transwell检测VSMCs迁移,q PCR检测miR-23b及其靶基因的PI3KR1、Smad3表达。结果 HG能浓度依赖性的促进VSMCs的增殖并抑制miR-23b的表达,与对照组比较差异均有统计学意义(P<0.05)。VSMC^(miR-23b)组的细胞活力及迁移率均显著低于VSMCNC组(P<0.05)。HG能升高VSMCNC及VSMC^(miR-23b)的细胞活力及迁移率,与对照组比较差异有统计学意义(P<0.05),VSMC^(miR-23b)+高糖组的细胞活力及迁移率均显著低于VSMCNC+高糖组(P<0.05)。VSMC^(miR-23b)组的PI3KR1、Smad3的表达显著低于VSMCNC组(P<0.05)。高糖能升高VSMCNC及VSMC^(miR-23b)的PI3KR1、Smad3的表达。VSMC^(miR-23b)+高糖组的PI3KR1、Smad3的表达均显著低于VSMCNC+高糖组(P<0.05)。结论 miR-23b可能通过调控靶基因Smad3、PI3KR1参与HG诱导的血管平滑肌增殖及迁移。

Objective High glucose induced vascular smooth muscle cell proliferation and migration is the key to atherosclerosis pathology caused by diabetes. The role of small RNA in diabetic vascular complications is taken seriously increasingly. This project will focus on the mi R-23 b on proliferation and migration induced by high glucose in vascular smooth muscle cell. Methods VSMCs were incubated with high glucose（HG,40 m M）for 24 h. The expression of mi R-23 b induced by high dose glucose was detected by q PCR. In order to further confirm the role of mi R-23 b on sugar induced VSMC proliferation and migration,slow virus technology was used to construct an over-express mi R-23 b VSMCs（VSMCmi R-23b） and control cells（VSMC^NC）. D-glucose incubate with VSMC^miR-23b and VSMC^NC respectively for 24 h,VSMCs proliferation was tested by CCK8,the migration of VSMCs was analyzed by Transwell,the expression of mi R-23 b and its target genes（PI3KR1,Smad3） were detected by q PCR. Results Compared with the control group,D-glucose can significant inhibit the expression of mi R-23 b with concentration-dependent（P〈0.05）. Cell vitality and mobility in VSMC^miR-23b group were significantly lower than VSMC^NC group（P〈0.05）. Compared with the control group,Glucose can significant raise the cell vitality and mobility in VSMC^miR-23b and VSMC^NC（P〈0.05）. The cell vitality and mobility of VSMCmi R-23b＋ high glucose group were significantly lower than VSMC^NC ＋ high glucose group（P〈0.05）. The expression of PI3KR1 and Smad3 in VSMC mi R-23 b group was significantly lower than VSMC^NC group（P〈0.05）. High glucose can raise the expression of PI3KR1 and Smad3 in VSMC mi R-23 b and VSMC^NC group. The expression of PI3KR1 and Smad3 in VSMC^miR-23b ＋ high glucose group were significantly lower than VSMC^NC ＋ high glucose group（P〈0.05）. Conclusionmi R-23 b may participate in proliferation and migration induced by high glucose in vascular smooth muscle through regulating the target genes PI3KR1,Smad3.