丹参总酚酸对慢性肾衰竭大鼠肾组织紧密连接蛋白ZO-1、occludin表达的影响  

The Effect of Total Salvianolic Acid on the Expression of Tight Junction Protein ZO-1 and Occluding in the Renal Fibrosis of Chronic Renal Failure Rats

作　　者：周萍[1] 马欣[1] 高芳[1] 樊均明[1] 毛楠[1] ZHOU Ping;MA Xin;GAO Fang(Nephrology Department,First Affiliated Hospital of Chengdu Medical College,Chengdu(610500))

机构地区：[1]成都医学院第一附属医院肾病科,成都610500

出　　处：《中国中西医结合肾病杂志》2022年第3期195-199,I0001,I0002,共7页Chinese Journal of Integrated Traditional and Western Nephrology

基　　金：四川省养老与老年健康协同创新中心项目(No.YLZBZ2023);成都市金牛区医学会肾脏专委会专项科研计划项目(No.JNQN20-01)。

摘　　要：目的:观察丹参总酚酸通过调控紧密连接蛋白ZO-1、occludin表达对慢性肾衰竭大鼠肾脏纤维化的作用。方法:将120只大鼠分为正常组(Nor)、模型组(Mod)、丹参总酚酸组低剂量组(TSA-L)、丹参总酚酸组中剂量组(TSA-M)、丹参总酚酸组高剂量组(TSA-H)及福辛普利组(Fos)。模型组,丹参总酚酸低、中、高剂量组,福辛普利组均通过腺嘌呤灌胃建立大鼠慢性肾衰竭模型,丹参总酚酸组建模后分别按照低、中、高剂量给予丹参总酚酸灌胃,福辛普利组大鼠建模后予以福辛普利灌胃,模型组及正常组大鼠同等体积生理盐水灌胃。所有大鼠于建模后给药2周后采集血液标本行血肌酐、尿素氮检测;处死前收集24 h尿液行24 h尿蛋白定量检测。收集肾脏组织行HE和Masson染色观察病理及纤维化情况分析,且分别行Real time-PCR、免疫组织化学及免疫印迹法(Western Blotting)测定肾脏紧密连接蛋白ZO-1、occludin mRNA及蛋白表达。结果:丹参总酚酸中、高剂量组和福辛普利组大鼠血肌酐、尿素氮水平较模型组降低(P<0.05);丹参总酚酸高剂量组与模型组比较,24 h尿蛋白水平明显降低,差异有统计学意义(P<0.05)。模型组大鼠肾脏组织病理评分及肾间质纤维化面积明显高于正常组(P<0.01),而丹参总酚酸中、高剂量组病理评分及纤维化面积低于模型组(P<0.05),与福辛普利组差异无统计学意义(P>0.05);模型组,丹参总酚酸低、中、高剂量组,福辛普利组的ZO-1、occludinm RNA和蛋白表达均显著低于正常组(P<0.05);而丹参总酚酸中、高剂量组,以及福辛普利组明显高于模型组(P<0.05),其中以丹参总酚酸高剂量组增加最为显著。结论:丹参总酚酸可能通过调控紧密连接蛋白ZO-1、occludin的表达,从而干预慢性肾衰竭肾纤维化的发生,延缓肾功能进展。Objective:To study the effect of total salvianolic acid on the renal fibrosis in chronic renal failure rats by regulating the expression of tight junction protein ZO-1 and occludin.Methods:120 rats were divided into six groups,which were normal(Nor)group,model(Mod)group,total salvianolic acid low dose(TSA-L)group,total salvianolic acid middle dose(TSA-M)group,total salvianolic acid high dose(TSA-H)group and Fosinopril(Fos)group.Rat models of chronic renal failure were established by intragastric adenine gavage in the Mod group,TSA-L group,TSA-M group,TSA-H group and TSA group.Rats in TSA group were fed with different total salvianolic acid for two weeks after modeling.Rats in Fos group were fed with Fosinopril for two weeks after modeling Model rats and normal rats received normal saline.Blood samples and urine were collected at 6 weeks later for measuring Scr、BUN and 24-hour urinary protein.Irrigation kidneys were harvested for HE and Masson staining at 6 weeks.The expression of ZO-1 and occludin were assessed with immunohistological staining and western blotting.The RT-PCR was employed to detect the ZO-1 and occludin mRNA levels in kidney samples.Results:The levels of serum creatinine and urea nitrogen in the middle and high dose groups of salvianolic acid and the fosinopril group were lower than those in the model group(P<0.05).Compared with the model group,the high-dose phenolic acid group of Danshen was significantly lower in 24-hour urine protein(P<0.05).The renal histopathological scores and renal interstitial fibrosis area were significantly higher in the model group.Compared with the normal group(P<0.01),the pathological score and fibrosis area of the salvia miltiorrhiza total and high dose group were lower than the model group(P<0.05),but there was no statistically significant difference with the fosinopril group(P>0.05);Model group,low-,medium-,and high-dose phenolic acid groups,ZO-1、occludin mRNA and protein expression in the fosinopril group were significantly lower than the normal group(P<0.05);

关 键 词：丹参总酚酸 慢性肾衰竭 肾纤维化 ZO-1 OCCLUDIN 

分 类 号：R285.5[医药卫生—中药学]