Chen Wei-Ren, Huo Xia, Zhou Yu-Jie, Chen Yun-Dai, Ma Qian, Jin Qin-Hua, Wu Xue-Ping, Sha Yuan
Department of Cardiology,Beijing Institute of Heart Lung and Blood Vessel Diseases,Beijing Key Laboratory of Precision Medicine of Coronary Atherosclerotic Disease,Clinical Center for Coronary Heart Disease of Capital Medical University,Beijing Anzhen Hospital,Capital Medical University,Beijing 100029,China.
Department of Cardiology,National Clinical Research Center for Geriatric Diseases,the Second Medical Center of Chinese PLA General Hospital,Beijing 100853,China.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2022 Jun;44(3):377-383. doi: 10.3881/j.issn.1000-503X.14421.
Objective To investigate the effect of dual-specificity phosphatase 1/optical atrophy 1 (DUSP1/OPA1) signaling pathway on vascular smooth muscle cell (VSMC) calcification.Methods An model of VSMC calcification was induced by exposure to β-glycerophosphate and calcium chloride.VSMC calcification was assessed by Alizarin Red S staining and calcium content by ELISA.Apoptosis was detected by TUNEL.Western blotting was employed to determine the protein levels of DUSP1,OPA1,Runt-related transcription factor 2 (Runx-2),bone morphogenetic protein 2 (BMP-2),and cysteinyl aspartate-specific proteinase-3 (Caspase-3).The effects of DUSP1 overexpression and OPA1 knockdown on cell calcification were investigated.Results Calcium chloride and β-glycerolphosphate induced VSMC calcification and down-regulated the expression levels of DUSP1 (=11.951,<0.001) and OPA1 (=8.487,<0.001).DUSP1 overexpression promoted OPA1 expression (=-8.921,<0.001),attenuated VSMC calcification,reduced calcium content and apoptosis rate,and down-regulated the expression of Runx-2,BMP-2,and active Caspase-3 (all <0.001).OPA1 knockdown increased calcium content and apoptosis rate,up-regulated the expression of Runx-2,BMP-2,and active Caspase-3,and promoted VSMC calcification (all <0.001).Conclusion DUSP1 may inhibit the VSMC calcification through the OPA1 signaling pathway.
目的 探讨双特异性磷酸酶1/视神经萎缩蛋白1（DUSP1/OPA1）信号通路对血管平滑肌细胞（VSMC）钙化的影响。方法 通过暴露于β-甘油磷酸和氯化钙诱导建立VSMC钙化模型。采用茜素红S染色评估VSMC钙化情况，用ELISA检测钙含量。通过TUNEL法检测细胞凋亡。采用蛋白质印迹法测定DUSP1、OPA1、 runt相关转录因子2（Runx-2）、骨形态发生蛋白2（BMP-2）和半胱天冬酶-3（Caspase-3）的蛋白水平。研究DUSP1过表达和OPA1敲低对细胞钙化的影响。结果 氯化钙和β-甘油磷酸诱导VSMC钙化，并下调DUSP1（=11.951，<0.001）和OPA1（=8.487，<0.001）的表达水平。DUSP1过表达促进OPA1表达（=-8.921，<0.001），减轻VSMC钙化，降低钙含量和凋亡率，并下调Runx-2、BMP-2和活性Caspase-3的表达（均<0.001）。OPA1敲低增加钙含量和凋亡率，上调Runx-2、BMP-2和活性Caspase-3的表达，并促进VSMC钙化（均<0.001）。结论 DUSP1可能通过OPA1信号通路抑制VSMC钙化。
