短期高盐喂食对C57BL/6J小鼠肾小管上皮细胞间充质转化和肾脏纤维化的影响  

Effects of short-term high salt diet on renal tubule epithelial mesenchymal transformation and renal fibrosis in C57BL/6J mice

作　　者：黄娟 商黔惠 陈春艳 HUANG Juan;SHANG Qian-hui;CHEN Chun-yan(Clinical Medicine Institute,Cardiovascular Diseases Institute,Hypertension Research Institute of Zunyi Medical University,Cardiology Department of Affiliated Hospital of Zunyi Medical University,Zunyi Guizhou 563003,China)

机构地区：[1]遵义医科大学附属医院临床医学研究所,心血管病研究所,高血压研究室,遵义医科大学附属医院心内科,贵州遵义563003

出　　处：《中华高血压杂志》2020年第8期757-766,共10页Chinese Journal of Hypertension

基　　金：国家自然科学基金(81160041、81460077);贵州省国际科技合作计划[G字(2012)7038号];贵州省优秀科技教育人才省长专项基金项目[黔省专合字(2012)15号]。

摘　　要：目的探讨短期高盐喂食对C57BL/6J小鼠肾小管上皮细胞间充质转化和肾脏纤维化的影响。方法雄性C57BL/6J小鼠20只,随机分为对照组(饲以0.5%NaCl颗粒饲料)和高盐组(饲以8%NaCl颗粒饲料),每组各10只。测小鼠尾动脉血压,1次/周,共3周;代谢笼收集各组小鼠24 h尿液;3周测定血肌酐、尿素氮、24 h尿微量白蛋白(MAU)及尿钠、钾含量,计算肌酐清除率(Ccr)及24 h尿钠/钾;采用苏木精-伊红(HE)、过碘酸-希夫(PAS)、Masson染色观察肾脏形态及纤维化,用免疫组化和Western blot法分别检测小鼠肾脏E-钙黏蛋白(E-cadherin)、波形蛋白(Vimentin)、α-平滑肌肌动蛋白(α-SMA)及肾脏损伤分子-1蛋白(Kim-1)的分布和表达量。结果实验3周后,两组小鼠血压比较差异无统计学意义(P>0.05);与对照组比,高盐组小鼠24 h MAU[(0.39±0.09)比(0.07±0.01)mg]、Ccr[(1.99±0.46)比(1.25±0.58)L/(g·min)]、24 h尿钠[(0.62±0.28)比(0.01±0.01)mmol]、24 h尿钾[(0.29±0.04)比(0.05±0.01)mmol]及尿钠/钾(4.31±1.80比0.08±0.03)增加(均P<0.05),肾小球硬化指数、肾小球纤维化指数及肾间质纤维化指数升高(均P<0.05)。免疫组化及Western blot结果显示,与对照组相比,高盐组小鼠肾脏E-cadherin蛋白表达减少,Vimentin、α-SMA及Kim-1蛋白表达增加(均P<0.05)。结论短期高盐喂食可能通过非血压依赖途径诱导正常血压C57BL/6J小鼠肾小管上皮细胞间充质转化的发生,促进肾脏纤维化。Objective To investigate the effects of short-term high salt diet on the interstitial transformation(EMT)of renal tubular epithelial cells and renal fibrosis in C57 BL6/J mice.Methods Twenty male C57 BL/6 J mice were randomly divided into control group(fed with 0.5%NaCl pellet feed)and high salt group(fed with 8%NaCl pellet feed),10 mice in each group.The tail artery blood pressure was measured once a week for 3 weeks.24-hour urine was collected by metabolic cage.The serum creatinine,urea nitrogen,24-hour urinary microalbumin(MAU),urinary sodium and potassium were measured 3 weeks after intervention.Creatinine clearance rate(Ccr)and 24-hour urinary sodium/potassium ratio were calculated.The renal morphology and fibrosis were observed by HE(hematoxylin-eosin),PAS(periodic acid-Schiff)and Masson staining.The protein distribution and expression of E-cadherin,Vimentin,α-smooth muscle actin(α-SMA)and kidney injury molecule-1(Kim-1)in the kidneys of mice was detected with immunohistochemistry and Western blot.Results After 3 weeks of high-salt diet,there was no significant difference in blood pressure among two groups(P>0.05).Compared with the control group,the 24-hour MAU[(0.39±0.09)vs(0.07±0.01)mg],Ccr[(1.99±0.46)vs(1.25±0.58)L/(g·min)],24-hour urinary sodium[(0.62±0.28)vs(0.01±0.01)mmol],24-hour urinary potassium[(0.29±0.04)vs(0.05±0.01)mmol]and 24-hour urinary sodium/potassium ratio(4.31±1.80 vs 0.08±0.03)increased in the high salt group(P<0.05).The glomerular sclerosis index,glomerular fibrosis index and renal interstitial fibrosis index were also increased(P<0.05).As compared with control,the expression of Vimentin,α-SMA and Kim-1 increased while E-cadherin decreased(all P<0.05)in the high salt as indicated by immunohistochemistry and Western blot analysis.Conclusion Short term high salt diet may induce renal tubule EMT and promote renal fibrosis in C57 BL/6 J mice with normal blood pressure through blood pressure independent pathway.

关 键 词：高盐 短期 C57BL/6J小鼠 肾脏纤维化 间充质转化 

分 类 号：R544.1[医药卫生—心血管疾病]