Jin Tianhui, Chen Liang, Liu Yehong, Sheng Ying, Zhou Yuting, Xuan Shiyi, Zong Gangjun
Department of Cardiology, Wuxi Clinical College, Anhui Medical University, Wuxi 214044; Dept of Cardiology, No. 904 Hospital of the PLA Joint Logistic Support Force, Wuxi 214044, China.
Department of Cardiology, Wuxi Clinical College, Anhui Medical University, Wuxi 214044, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2021 Dec;37(12):1079-1084.
Objective To investigate the protective effect of proprotein convertase subtilisin/kexin type 9 (PCSK9) knockdown on endothelial-to-mesenchymal transition (EndoMT) induced by β glycerophosphate, dexamethasone, and L-ascorbic acid in human aortic endothelial cells (HAECs). Methods EndoMT model was established by inducing HAECs with (0, 10, 30, 50) mmol/L of β glycerophosphate combined with 100 nmol/L of dexamethasone and 50 μg/mL of L-ascorbic acid. HAECs were transfected with specific small interfering RNA of PCSK9 (si-PCSK9), and the mRNA and protein expression levels of PCSK9 in HAECs were detected by real-time quantitative PCR and Western blotting. HAECs were randomized into blank group, EndoMT group, EndoMT group transfected with negative control small interfering RNA (si-NC), and EndoMT group transfected with si-PCSK9. The mRNA levels of α-smooth muscle actin (α-SMA), fibroblast-specific protein 1 (FSP1), and vascular endothelial cadherin (VE-cadherin) were detected by real-time quantitative PCR, the protein levels of α-SMA and VE-cadherin were detected by Western blotting, and the expression of α-SMA was detected by immunofluorescence staining. Results 30 mmol/L of β glycerophosphate had the best effect in inducing EndoMT, and the expression of PCSK9 mRNA and protein was up-regulated when EndoMT occurred. After PCSK9 knockdown, the expressions of α-SMA and FSP1 were down-regulated, while the expression of VE cadherin was up-regulated. Conclusion Knockdown of PCSK9 inhibits the EndoMT of HAECs.
目的 探讨沉默前蛋白转化酶枯草溶菌素9型（PCSK9）对β-甘油磷酸、地塞米松和L-抗坏血酸诱导的人主动脉内皮细胞（HAECs）内皮-间充质转化（EndoMT）的保护作用。方法 用（0、10、30、50）mmol/L的β-甘油磷酸联合100 nmol/L地塞米松和50 μg/mL L-抗坏血酸诱导HAECs建立EndoMT模型。用PCSK9特异性小干扰RNA（si-PCSK9）转染HAECs，通过实时定量PCR和蛋白质印迹法检测HAECs中PCSK9的mRNA和蛋白表达水平。将HAECs随机分为空白组、EndoMT组、转染阴性对照小干扰RNA（si-NC）的EndoMT组和转染si-PCSK9的EndoMT组。通过实时定量PCR检测α-平滑肌肌动蛋白（α-SMA）、成纤维细胞特异性蛋白1（FSP1）和血管内皮钙黏蛋白（VE-cadherin）的mRNA水平，通过蛋白质印迹法检测α-SMA和VE-cadherin的蛋白水平，通过免疫荧光染色检测α-SMA的表达。结果 30 mmol/L的β-甘油磷酸诱导EndoMT效果最佳，EndoMT发生时PCSK9的mRNA和蛋白表达上调。沉默PCSK9后，α-SMA和FSP1的表达下调，而VE-cadherin的表达上调。结论 沉默PCSK9可抑制HAECs的EndoMT。
