Cer(d18:1/16:0)促进HK-2细胞线粒体障碍和纤维化表型转换  

Cer(d18:1/16:0)Promotes Mitochondrial Dysfunction and Fibrotic Phenotype Transformation in HK-2 Cells

作　　者：董政委 杨康 郭晓红 赵换 DONG Zhengwei;YANG Kang;GUO Xiaohong;ZHAO Huan(The First Affiliated Hospital of Henan University of Chinese Medicine,Heart Center/National Regional(Traditional Chinese Medicine)Cardiovascular Diagnosis and Treatment Center,Zhengzhou 450000,China;The First Affiliated Hospital of Henan University of Chinese Medicine,Department of Nephrology,Zhengzhou 450000,China;The First Affiliated Hospital of Shandong First Medical University(Shandong Province Qianfoshan Hospital),Jinan 250014,China;Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China)

机构地区：[1]河南中医药大学第一附属医院心脏中心/国家区域(中医)心血管诊疗中心,郑州450000 [2]河南中医药大学第一附属医院肾病科,郑州450000 [3]山东第一医科大学第一附属医院(山东省千佛山医院),济南250014 [4]天津中医药大学,天津301617

出　　处：《世界科学技术-中医药现代化》2024年第10期2545-2552,共8页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：河南省卫生健康委国家中医临床研究基地科研专项(2022JDZX109):基于肾小管上皮细胞外泌体调控的PPARγ信号通路探讨补阳还五汤干预肾间质纤维化的作用机制,负责人:杨康。

摘　　要：目的探究糖尿病肾病(Diabetic kidney disease,DKD)发展进程中的核心标志物Cer(d18:1/16:0)对人肾小管上皮细胞(Human renal proximal tubule,HK-2)线粒体功能障碍和纤维化表型转换的影响。方法借助qRT-PCR技术,检测高糖刺激下HK-2细胞神经酰胺合成的相关酶在转录水平的表达;使用CCK8法检测不同浓度Cer(d18:1/16:0)对HK-2细胞活力的影响;运用线粒体荧光探针评估不同浓度Cer(d18:1/16:0)刺激后HK-2细胞内线粒体数目变化;Western blot检测给予Cer(d18:1/16:0)前后线粒体相关蛋白及纤维化指标的表达情况。结果高糖环境下HK-2细胞神经酰胺合成酶2(Ceramide synthase 2,CERS2)表达增加;外源性Cer(d18:1/16:0)可显著抑制HK-2细胞活力;外源性Cer(d18:1/16:0)可降低HK-2细胞中线粒体总荧光强度和平均荧光强度;外源性Cer(d18:1/16:0)显著降低CV-ATP5A与CIII-UQCRC2的蛋白表达水平,显著增加Coll 3、α-SMA的蛋白表达水平。结论Cer(d18:1/16:0)可诱导HK-2细胞发生线粒体功能障碍与纤维化表型转换。Objective To explore the effect of Cer(d18:1/16:0),the core marker in the development of diabetic kidney disease(DKD),on mitochondrial dysfunction and phenotypic transformation of human renal proximal tubule cells(HK-2)induced by high glucose.Methodss With the help of qRT-PCR technology,the expression of ceramide synthesis related enzymes in HK-2 cells stimulated by high glucose was detected at the transcription level.The effects of different concentrations of Cer(d18:1/16:0)on the viability of HK-2 cells were detected by CCK8 method.The changes of mitochondrial number in HK-2 cells stimulated by different concentrations of Cer(d18:1/16:O)were evaluated by mitochondrial fluorescent probe.Western blot was used to detect the expression of mitochondrial related proteins and fibrosis indexes before and after the administration of Cer(d18:1/16:0).Results The expression of Ceramide synthase 2,(CERS2)in HK-2 cells increased in high glucose environment.Exogenous Cer(d18:1/16:0)can significantly inhibit the viability of HK-2 cells,can reduce the total and average fluorescence intensity of mitochondria in HK-2 cells induced by high glucose.The protein expression of CV-ATP5A and CIII-UQCRC2 decreased,while the protein expression of Coll 3,MMP9 and α-SMA increased.Conclusion Cer(d18:1/16:0)can induce mitochondrial dysfunction and fibrotic phenotype transformation in HK-2 cells stimulated by high glucose.

关 键 词：糖尿病肾病 Cer(d18:1/16:0) HK-2细胞 纤维化表型转换 线粒体功能障碍 

分 类 号：R587.2[医药卫生—内分泌]