Zhang Danhuan, Xi Minhui, Chen Lingyun, Huang Yanping, Mao Peiju
Department of Nephrology, Tongren Hospital Affiliated to Shanghai Jiaotong University School of Medicine, Shanghai 200336, P.R. China.
Department of Nephrology, Pudong New Area People's Hospital, Shanghai 201200, P.R. China.
Exp Ther Med. 2018 Feb;15(2):1193-1198. doi: 10.3892/etm.2017.5521. Epub 2017 Nov 16.
The present study aimed to investigate the effect of pentraxin 3 (PTX3) on the regulation of proliferation and apoptosis in human glomerular mesangial cells (HMCs). Small interfering (si)RNA was designed and synthesized to inhibit the expression of endogenous PTX3, and the effects on the proliferation and apoptosis of HMCs were detected by flow cytometry and an MTT assay. Western blot analysis was used to detect the activation of mitogen-activated protein kinase (MAPK) proteins in HMCs with PTX3 knockdown. Three siRNAs targeting PTX3 were individually transfected into HMCs for 48 h, and reverse-transcription quantitative PCR demonstrated that the relative mRNA expression of PTX3 was significantly decreased in all groups by up to 79.62% of that in the control group (P<0.05). Following transfection with PTX3-siRNA, the viability of an HMC line was significantly decreased in comparison with that of a control group transfected with scrambled siRNA. However, PTX3-siRNA did not significantly effect early and late apoptotic cell populations in HMCs compared with those in the control. Endogenous PTX3 interference was found to significantly decrease p38 MAPK, extracellular signal-regulated kinase 1/2 and c-Jun N-terminal kinase phosphorylation. In conclusion, silencing of PTX3, inhibited the proliferation of HMCs via MAPK pathways, but exerted no effect on the apoptosis of HMCs.
本研究旨在探讨五聚体3（PTX3）对人肾小球系膜细胞（HMCs）增殖和凋亡调控的影响。设计并合成小干扰（si）RNA以抑制内源性PTX3的表达，通过流式细胞术和MTT法检测其对HMCs增殖和凋亡的影响。采用蛋白质免疫印迹分析检测PTX3基因敲低的HMCs中丝裂原活化蛋白激酶（MAPK）蛋白的激活情况。将三种靶向PTX3的siRNA分别转染至HMCs中48小时，逆转录定量PCR结果显示，所有组中PTX3的相对mRNA表达均显著降低，最多可降至对照组的79.62%（P<0.05）。转染PTX3-siRNA后，与转染乱序siRNA的对照组相比，HMC系的活力显著降低。然而，与对照组相比，PTX3-siRNA对HMCs早期和晚期凋亡细胞群体无显著影响。发现内源性PTX3干扰可显著降低p38 MAPK、细胞外信号调节激酶1/2和c-Jun氨基末端激酶的磷酸化水平。总之，PTX3基因沉默通过MAPK途径抑制HMCs的增殖，但对HMCs的凋亡无影响。
