Meng Zheying, Wang Yu, Lin Yanduan, Nan Shuliang, Xu Weiping, Hu Bing, Shen E
Department of Ultrasound in Medicine, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai Institute of Ultrasound in Medicine, Shanghai 200233, China.
Department of Ultrasound in Medicine, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai Institute of Ultrasound in Medicine, Shanghai 200233, China; Email:
Zhonghua Xin Xue Guan Bing Za Zhi. 2015 Jul;43(7):619-24.
To investigate the role and signalling of microRNA(miR)-182 on regulating high glucose-induced cardiomyocyte hypertrophy.
The candidates of miR which might potentially be involved on targeting Rac1 were predicted by applying bioinformatics analysis. The expression of all related candidates miRs was verified by real-time reverse transcription-PCR (RT-PCR) in cardiac tissues of db/db mice and db/m mice. Then the relationship between candidates miR and Rac1 was investigated with Pearson relevant analysis. Neonatal mice cardiomyocytes were cultured and divided into 2 groups: normal glucose group and high glucose group. The level of selected miR and Rac1 in two groups was detected by RT-PCR. Neonatal mice cardiomyocytes were then randomly divided into 4 groups: normal glucose group, selected microRNA mimics control group, high glucose group, high glucose plus selected miR mimics control group. The morphology of cardiomyocyte in each group was detected under light microscope. Furthermore, Rac1, β-MHC and α-SMA expressions were detected in cultured cardiomyocyte treated by high glucose for 48 h after transfecting selected miR mimics by RT-PCR and Western blot.
A total of 6 miR candidates potentially targeting Rac1 were screened by bioinformatics, which were miR-182, miR-142-3p, miR-140, miR-101a, miR-429 and miR-200b. Among these candidates, miR-182 and miR-142-3p expression was significantly downregulated in cardiac tissues of db/db mice compared with db/m controls (P < 0.05). MiR-182 was negatively correlated with Rac1 by person analysis (r = -0.891 02). Downregulation of miR-182 and upregulation of Rac1, β-MHC, α-SMA were found in high glucose-induced cardiomyocyte. After transfection of miR-182 mimics, hypertrophic changes were significantly reduced and Rac1 as well β-MHC expression was significantly downregulated in cardiomyocyte incubated with high glucose.
MiR-182 might be involved in the regulation of high glucose-induced myocardial hypertrophy process via targeting Rac1.
研究微小RNA（miR）-182在调节高糖诱导的心肌细胞肥大中的作用及信号传导。
应用生物信息学分析预测可能靶向Rac1的miR候选物。通过实时逆转录聚合酶链反应（RT-PCR）验证db/db小鼠和db/m小鼠心脏组织中所有相关候选miR的表达。然后用Pearson相关分析研究候选miR与Rac1之间的关系。培养新生小鼠心肌细胞并分为2组：正常葡萄糖组和高糖组。通过RT-PCR检测两组中所选miR和Rac1的水平。然后将新生小鼠心肌细胞随机分为4组：正常葡萄糖组、所选微小RNA模拟物对照组、高糖组、高糖加所选miR模拟物对照组。在光学显微镜下检测每组心肌细胞的形态。此外，在转染所选miR模拟物后，通过RT-PCR和蛋白质免疫印迹法检测高糖处理48小时的培养心肌细胞中Rac1、β-肌球蛋白重链（β-MHC）和α-平滑肌肌动蛋白（α-SMA）的表达。
通过生物信息学共筛选出6个可能靶向Rac1的miR候选物，即miR-182、miR-142-3p、miR-140、miR-101a、miR-429和miR-200b。在这些候选物中，与db/m对照组相比，db/db小鼠心脏组织中miR-182和miR-142-3p的表达显著下调（P<0.05）。经Person分析，miR-182与Rac1呈负相关（r=-0.891 02）。在高糖诱导的心肌细胞中发现miR-182下调以及Rac1、β-MHC、α-SMA上调。转染miR-182模拟物后，在高糖孵育的心肌细胞中肥大变化显著减少，Rac1以及β-MHC表达显著下调。
MiR-182可能通过靶向Rac1参与高糖诱导的心肌肥大过程的调节。
