基于ENaC泛素化探讨补阳还五汤调控HK2细胞钠代谢作用机制  

Study on the mechanism of Buyang Huanwu Decoction regulating sodium metabolism in HK2 cells based on ENaC ubiquitination

作　　者：王建波[1] 曲怡[1] 孙杰 薛亚楠[1] 马永钢 张曼婷 张立德[1] WANG Jian-bo;QU Yi;SUN Jie;XUE Ya-nan;MA Yong-gang;ZHANG Man-ting;ZHANG Li-de(Liaoning University of Traditional Chinese Medicine,Innovative Engineering Technology Center of Traditional Chinese Medicine,Key Laboratory of Ministry of Education for TCM Viscera-State Theory and Applications,She-n yang110847,China)

机构地区：[1]辽宁中医药大学中医药创新工程技术中心,中医脏象理论及应用教育部重点实验室,辽宁沈阳110847

出　　处：《时珍国医国药》2022年第11期2627-2630,共4页Lishizhen Medicine and Materia Medica Research

基　　金：辽宁省教育厅育苗项目(L202025);辽宁省百千万人才工程项目(2020921097);国家自然科学基金(81302880);国家重点基础研究发展计划(973计划)项目(2012CB518503)。

摘　　要：目的 研究补阳还五汤调控上皮钠通道(ENaC)泛素化,保护人肾小管上皮细胞(HK2)钠代谢异常作用机制。方法 采用高盐(150mmol/L)干预HK2细胞24h,诱导细胞损伤。加入不同剂量的补阳还五汤(0.5%,1%,2%)干预24h,收集细胞。用Hoechst33342染色,高内涵观测细胞数量;荧光定量PCR检测SGK1、NEDD4-2、ENaC mRNA含量;Wes全自动蛋白表达分析系统检测SGK1、NEDD4-2、ENaC的蛋白表达。结果 高盐组造模后,细胞数量显著减少,细胞活性变差;SGK1、ENaC mRNA含量及蛋白表达显著升高(P<0.05);NEDD4-2 mRNA及蛋白含量显著降低(P<0.05)。补阳还五汤干预治疗,可以显著增加细胞数量及活性,抑制SGK1表达,增加泛素连接酶NEDD4-2含量,促进ENaC泛素化降解,从而减少HK2细胞钠离子重吸收,差异具有统计学意义(P<0.05)。结论 补阳还五汤能激活HK2细胞泛素化系统,通过促进ENaC泛素化降解,减少细胞钠离子重吸收,从而起到盐敏感高血压肾脏保护作用。Objective To study the mechanism of Buyang Huanwu Decoction in regulating the ubiquitination of epithelial sodium channel(ENaC) and protecting the abnormal sodium metabolism of human renal tubular epithelial cells(HK2).Methods High salt(150mmol/L) was used to intervene HK2 cells for 24h to induce cell injury. Different doses of Buyang Huanwu Decoction(0.5%, 1%, 2%) were added for 24 hours to collect cells. Hoechst33342 staining was used to observe the number of cells;The mRNA contents of SGK1, Nedd4-2 and ENaC were detected by fluorescence quantitative PCR;The protein expressions of SGK1, Nedd4-2 and ENaC were detected by WES automatic protein expression analysis system.Results In the high salt group, the number of cells decreased significantly and the activity of cells became worse;The mRNA content and protein expression of SGK1 and ENaC increased significantly(P<0.05);The contents of Nedd4-2 mRNA and protein decreased significantly(P<0.05). Buyang Huanwu Decoction can significantly increase the number and activity of cells, inhibit the expression of SGK1, increase the content of ubiquitin ligase Nedd4-2, promote the ubiquitination and degradation of ENaC, and reduce the sodium reabsorption of HK2 cells(P<0.05). Conclusion Buyang Huanwu Decoction can activate the ubiquitination system of HK2 cells, promote the degradation of ENaC ubiquitination and reduce the reabsorption of sodium ions, so as to protect the kidney of salt sensitive hypertension.

关 键 词：补阳还五汤 HK2 高盐 泛素化 ENAC 

分 类 号：R285.5[医药卫生—中药学]