Shen P F, Qu Y X, Wang B, Xu J D, Wei K, Xie Z K, Ma Y
Department of Orthopedics, Changzhou Traditional Chinese Medicine Hospital Affiliated to Nanjing University of Chinese Medicine, Changzhou 213004, China.
Zhonghua Yi Xue Za Zhi. 2017 Oct 24;97(39):3079-3084. doi: 10.3760/cma.j.issn.0376-2491.2017.39.008.
To investigate the expression of miR-30a-5p in cartilage of osteoarthritis patients, and to explore its mechanism of chondrocyte apoptosis. From May 2015 to December 2016, tissue specimen of 289 patients with osteoarthritis was collected in Department of Orthopedics, Changzhou traditional Chinese Medicine Hospital Affiliated to Nanjing University of Chinese Medicine.The expression of miR-30a-5p and protein kinase B(Akt) mRNA in cartilage of different patients was detected by qPCR.The apoptosis of chondrocytes was detected by Tunel method.The expression of related proteins in tissues and cells was detected by immunoblotting, and apoptosis and cell cycle were detected by flow cytometry. The expression of miR-30a-5p in OA patients was significantly higher than control patients (<0.05), but Aktwas positively related[(3.64±0.95)vs(1.03±0.31), <0.05]. The expression of miR-30a-5p in cartilage of OA patients was negatively correlated with Akt mRNA expression (=0.729 3, <0.001), but it had a positive correlation with the apoptotic rate (=0.847 5, <0.001). miR-30a-5p targets negative regulation of Akt gene expression in SW1353 cells, and the expression of p-Akt, IkB-α, p-IkB-α, p65, p-p65 and mTOR and p-mTOR were significantly down-regulated by miR-30a-5p (<0.05). Compared with normal SW1353 cells, the apoptosis rate of SW1353 cells which was transfected with miR-30a-5p-mimics increased by 9.65 times, G0/G1 phase cells increased by 1.37 times, S phase cells decreased by 60.94%, G2/M phase cells decreased 19.53%. miR-30a-5p is highly expressed in cartilage of osteoarthritis patients, and its high expression can block chondrocytes in G0/G1 phase by targeting Akt gene, and induce apoptosis of chondrocytes.
探讨骨关节炎患者软骨中miR-30a-5p的表达情况，并探究其诱导软骨细胞凋亡的机制。2015年5月至2016年12月，南京中医药大学附属常州中医医院骨科收集了289例骨关节炎患者的组织标本。采用qPCR检测不同患者软骨中miR-30a-5p和蛋白激酶B（Akt）mRNA的表达。采用Tunel法检测软骨细胞的凋亡情况。采用免疫印迹法检测组织和细胞中相关蛋白的表达，采用流式细胞术检测细胞凋亡和细胞周期。骨关节炎患者中miR-30a-5p的表达显著高于对照组患者（<0.05），但Akt呈正相关[（3.64±0.95）vs（1.03±0.31），<0.05]。骨关节炎患者软骨中miR-30a-5p的表达与Akt mRNA表达呈负相关（=0.729 3，<0.001），但与凋亡率呈正相关（=0.847 5，<0.001）。miR-30a-5p在SW1353细胞中靶向负调控Akt基因表达，miR-30a-5p可使p-Akt、IkB-α、p-IkB-α、p65、p-p65和mTOR以及p-mTOR的表达显著下调（<0.05）。与正常SW1353细胞相比，转染miR-30a-5p模拟物的SW1353细胞凋亡率增加了9.65倍，G0/G1期细胞增加了1.37倍，S期细胞减少了60.94%，G2/M期细胞减少了19.53%。miR-30a-5p在骨关节炎患者软骨中高表达，其高表达可通过靶向Akt基因使软骨细胞阻滞于G0/G1期，并诱导软骨细胞凋亡。
