尿毒症患者血清通过ROS-NLRP3信号通路促进人主动脉平滑肌细胞增殖参与新生内膜增生  

Uremia Serum Promotes the Proliferation of Human Aortic Smooth Muscle Cells through the ROS-NLRP3 Signaling Pathway and Participates in Neointimal Hyperplasia

作　　者：段琦 李亚峰 王振峰[6] DUAN Qi;LI Yafeng;WANG Zhenfeng(Laboratory of Molecular Diagnosis and Therapy of Kidney Disease,Shanxi Provincial People's Hospital,Taiyuan 030012,Shanxi,China;Shanxi Provincial Key Laboratory of Kidney Disease,Shanxi Provincial People's Hospital,Taiyuan 030012,Shanxi,China;Department of Nephrology,Shanxi Provincial People's Hospital,Taiyuan 030012,Shanxi,China;Core Laboratory,Shanxi Provincial People's Hospital,Taiyuan 030012,Shanxi,China;Academy of Microbial Ecology,Shanxi Medical University,Taiyuan 030012,Shanxi,China;Department of Vascular Surgery,Shanxi Provincial People's Hospital,Taiyuan 030012,Shanxi,China)

机构地区：[1]山西省人民医院肾脏病分子诊断与治疗实验室,太原030012 [2]山西省人民医院肾脏病山西省重点实验室,太原030012 [3]山西省人民医院肾内科,太原030012 [4]山西省人民医院中心实验室,太原030012 [5]山西医科大学微生态研究院,太原030012 [6]山西省人民医院血管外科,太原030012

出　　处：《中国分子心脏病学杂志》2024年第3期6110-6115,共6页Molecular Cardiology of China

基　　金：山西省自然科学基金面上自然基金项目(201901D111439);山西省基础研究自由探索类青年科学研究项目(20210302124303)。

摘　　要：目的通过在体外模拟慢性肾脏病(chronic kidney disease,CKD)病理环境探讨ROS-NLRP3信号通路在CKD中对血管新生内膜增生的影响。方法体外培养人主动脉平滑肌细胞(human aortic smooth muscle cells,HASMCs),实验分为3组:正常血清对照组(NC组)、尿毒症血清处理组(CKD组)、尿毒症血清+Mito-TEMPO处理组(CKD+Mito-TEMPO组)。采用Western blot、免疫荧光和ELISA实验方法观察尿毒症血清对HASMCs表型、氧化应激水平、炎症因子表达水平以及NLRP3炎症复合体表达的影响。结果与NC组相比,经尿毒症患者血清处理后,HASMCs增殖显著增强,平滑肌肌动蛋白α(smooth muscle actinα,α-SMA)和钙调理蛋白1的蛋白表达、线粒体活性氧(mitochondrial reactive oxygen species,mtROS)水平显著增加(P均<0.05),炎症因子白细胞介素-6(interleukin-6,IL-6)、IL-18和TNF-α的水平显著增加(P均<0.05),nod样受体蛋白3(nod-like receptor protein 3,NLRP3)、半胱氨酸天冬氨酸蛋白水解酶-1(cysteinyl aspartate specific proteinase-1,caspase-1)、剪切的caspase-1、IL-1β和IL-18的蛋白表达水平显著增加(P均<0.05),超氧化物歧化酶(superoxide dismutase,SOD)活性降低显著降低(P<0.05);而加入mtROS抑制剂后,HASMCs增殖显著降低;α-SMA和钙调理蛋白1的蛋白表达,mtROS水平,炎症因子IL-6、IL-18、TNF-α水平,NLRP3、caspase-1、剪切的caspase-1、IL-1β和IL-18蛋白表达水平均显著减少(P均<0.05);SOD活性降低显著增加(P<0.05)。结论尿毒症血清通过ROS-NLRP3信号通路促进HASMCs增殖、参与新生内膜增生。Objective To explore the effect of ROS-NLRP3 signaling pathway on neointimal hyperplasia(NH)induced by chronic kidney disease(CKD)in vitro.Methods Human aortic smooth muscle cells(HASMCs)were cultured and divided into three groups:normal serum control(NC)group,uremia serum treatment group,uremia serum+Mito-TEMPO treatment group.The effects of uremia serum on HASMCs phenotype,oxidative stress level,expression levels of inflammatory factors and non-like receptor protein 3(NLRP3)inflammatory complex were detected by Western blot,immunofluorescence and ELISA assays.Results Compared to NC group,the proliferation of HASMCs,the protein expressions ofα-SMA and calponin 1,the levels of mitochondrial reactive oxygen species(mtROS),and inflammatory factor IL-6,IL-18 and TNF-α,protein expressions of NLRP3,cysteinyl aspartate specific proteinase-1(caspase-1),cleaved caspase-1,IL-1β,and IL-18 were significantly increased(allP<0.05),superoxide dismutase(SOD)activity was significantly decreased(allP<0.05)after treatment of uremia serum.However,the proliferation of HASMCs,the protein expressions ofα-SMA and calponin 1,levels of mtROS,inflammatory factor IL-6,IL-18 and TNF-α,and the protein expression levels of NLRP3,caspase-1,cleaved caspase-1,IL-1β,and IL-18 were significantly decreased(allP<0.05),SOD activity was significantly increased(P<0.05)in uremia serum+Mito-TEMPO treatment group.Conclusion Uremia serum can promote the proliferation of HASMCs in neointimal hyperplasia through ROS-NLRP3 signaling pathway.

关 键 词：尿毒症 新生内膜增生 线粒体活性氧 人主动脉平滑肌细胞 NLRP3 

分 类 号：R692.5[医药卫生—泌尿科学]