WNK463缓解高渗透压状态下心肌细胞炎症反应和线粒体功能障碍的研究  

WNK463 alleviates cardiomyocytes inflammation and mitochondrial dysfunction under hypertonic stress

作　　者：焦新新 赵培源 曹源 Jiao Xinxin;Zhao Peiyuan;Cao Yuan(Department of Basic Medical Sciences,The 960th Hospital of the PLA Joint Logistics Support Force,Jinan 250031,China;Sun Yat-sen University Zhongshan School of Medicine,Guangzhou 510080,China)

机构地区：[1]解放军联勤保障部队第九六〇医院基础医学实验室,济南250031 [2]中山大学中山医学院,广州510080

出　　处：《中国临床实用医学》2025年第1期44-51,共8页China Clinical Practical Medicine

基　　金：山东省医药卫生科技发展计划项目(202102051099)。

摘　　要：目的探讨无赖氨酸激酶463(WNK463)在高渗透压状态下对心肌细胞炎症反应和线粒体功能障碍的缓解作用。方法选取AC16人心肌细胞作为实验模型,对细胞进行不同浓度的山梨醇处理,并将不同浓度的山梨醇与WNK463进行联合处理。采用细胞计数试剂盒-8检测细胞的存活率,通过实时定量聚合酶链式反应(RT-qPCR)检测细胞炎症因子和线粒体功能基因的表达,并采用免疫荧光染色技术观察无赖氨酸激酶1(WNK1)的荧光信号及线粒体形态变化。结果山梨醇处理AC16人心肌细胞12 h后,50、100 mmol/L山梨醇处理细胞的存活率[(72.06±1.96)%、(62.37±8.88)%]均低于未处理的AC16人心肌细胞[(99.11±4.20)%],差异有统计学意义(P<0.05);山梨醇处理24 h后,50、100 mmol/L山梨醇处理细胞的存活率[(86.00±3.19)%、(64.70±1.49)%]均低于未处理的AC16人心肌细胞[(98.00±4.33)%],差异有统计学意义(P<0.05)。采用50 mmol/L山梨醇处理细胞的白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、肿瘤坏死因子α(TNF-α)表达水平[(3.51±0.23)、(2.21±0.06)、(2.56±0.39)]与100 mmol/L山梨醇处理细胞的IL-1β、IL-6、TNF-α表达水平[(10.07±1.42)、(4.61±0.59)、(12.02±1.58)],均高于未处理的AC16人心肌细胞[(1.09±0.03)、(0.96±0.04)、(0.99±0.03)],差异有统计学意义(P<0.05);采用100 mmol/L山梨醇处理细胞的白细胞介素-8(IL-8)的表达水平(25.3±0.49)高于未处理的AC16人心肌细胞(1.06±0.09),差异有统计学意义(P<0.05)。采用50 mmol/L山梨醇处理细胞的磷酸酶及张力蛋白同源物诱导激酶1(PINK1)、过氧化物酶体增殖物激活受体γ共激活剂1α(PGC-1α)表达水平[(0.27±0.01)、(0.30±0.03)]与100 mmol/L山梨醇处理细胞的PINK1、PGC-1α表达水平[(0.34±0.05)、(0.25±0.05)],均低于未处理的AC16人心肌细胞[(1.02±0.09)、(1.09±0.18)],山梨醇处理12、24 h后,50 mmol/L山梨醇+WNK463处理细胞的存活率[(84.86±5.80)%、(96.07±1.64)%]均高于50 ObjectiveTo investigate the effect of with no lysine kinase 463(WNK463)on cardiomyocytes inflammation and mitochondrial dysfunction under hypertonic stress.MethodsAC16 human cardiomyocytes were selected as the experimental model,and the cells were treated with sorbitol at different concentrations,and sorbitol was combined with WNK463.The survival rate of the cells was detected by cell counting kit-8 method.Real-time quantitative polymerase chain reaction was used to detect the expression of inflammatory factors and mitochondrial functional genes,and immunofluorescence staining was used to observe the fluorescence signal of with no lysine kinase 1(WNK1)and the dynamic changes of mitochondrial morphology.ResultsAfter treating AC16 human cardiomyocytes with sorbitol for 12 h,the survival rates of 50 and 100 mmol/L sorbitol treated cells[(72.06±1.96)%,(62.37±8.88)%]were lower than those of untreated AC16 human cardiomyocytes[(99.11±4.20)%],the differences were statistically significant(P<0.05).After treatment with sorbitol for 24 h,the survival rates of 50 and 100 mmol/L sorbitol treated cells[(86.00±3.19)%,(64.70±1.49)%]were lower than those of untreated AC16 human cardiomyocytes[(98.00±4.33)%],and the differences were statistically significant(P<0.05).The expression levels of interleukin-1β(IL-1β),interleukin-6(IL-6),tumor necrosis factorα(TNF-α)in cells treated with 50 mmol/L sorbitol[(3.51±0.23),(2.21±0.06),(2.56±0.39)]and the expression levels of IL-1β,IL-6 and TNF-αin cells treated with 100 mmol/L sorbitol[(10.07±1.42),(4.61±0.59),(12.02±1.58)],were higher than those of untreated AC16 human cardiomyocytes[(1.09±0.03),(0.96±0.04),(0.99±0.03)],and the differences were statistically significant(P<0.05).The expression level of interleukin-8(IL-8)in 100 mmol/L sorbitol treated cells(25.3±0.49)was higher than that in untreated AC16 human cardiomyocytes(1.06±0.09),and the difference was statistically significant(P<0.05).The expression levels of phosphatase and tensin homolog induced putative kin

关 键 词：心肌细胞 无赖氨酸激酶463 无赖氨酸激酶1 山梨醇 高渗透压 

分 类 号：R542.2[医药卫生—心血管疾病]