绿原酸对粪肠球菌致小鼠肾损伤与高血压的作用  

Effects of Chlorogenic Acid on Enterococcus faecalis-Induced Renal Injury and Hypertensive in Mice

作　　者：刘裕婷 陶玉凤 曾钰婷 曾亮瑜 吕荣杰 陈扬[1] 王蕾 LIU Yuting;TAO Yufeng;ZENG Yuting;ZENG Liangyu;LYU Rongjie;CHEN Yang;WANG Lei(School of Pharmaceutical Sciences,Guangzhou University of Chinese Medicine,Guangzhou 510006Guangdong,China;Guangzhou Chuangtian BioTech Co.,Ltd,Guangzhou 511400 Guangdong,China)

机构地区：[1]广州中医药大学中药学院,广东广州510006 [2]广州创天生物科技有限公司,广东广州511400

出　　处：《中药新药与临床药理》2023年第1期64-71,共8页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：广东省自然科学基金杰出青年项目(2018B030306027);广州市科技重点项目(202103000044)。

摘　　要：目的 探究绿原酸(Chlorogenic acid,CGA)对粪肠球菌(Enterococcus faecalis,E.faecalis)诱导的高血压模型小鼠降压和肾脏保护作用。方法 将20只6~8周龄C57BL/6小鼠随机分为对照组、粪肠球菌组、绿原酸组和粪肠球菌+绿原酸组。给小鼠灌胃109CFU粪肠球菌或100 mg·kg^(-1)绿原酸溶液8周,期间使用尾套法测量血压,取材前使用代谢笼收集尿液。实验结束后心脏采血并收集肾脏。通过试剂盒检测尿液和血清的尿素氮、钠离子浓度,通过ELISA法检测尿液白蛋白含量,通过RT-qPCR法检测转化生长因子(TGF)-β、Collagen-1、NLRP3、白细胞介素(IL)-1β、IL-10、肿瘤坏死因子(TNF)-α和上皮钠离子通道(Epithelial sodium channel,ENaC)的转录水平以及粪便E.faecalis的相对含量,通过Western Blot法检测α-ENaC蛋白水平;通过紫外分光法检测菌液E.faecalis相对浓度。结果 与对照组比较,粪肠球菌组收缩压(Systolic blood pressure,SBP)、尿白蛋白、尿液尿素氮、肾脏损伤因子mRNA、血钠水平和α-ENaC表达明显升高(P<0.05,P<0.01);尿钠、IL-10转录水平明显降低(P<0.05)。与粪肠球菌组比较,粪肠球菌+绿原酸组的收缩压、24 h尿白蛋白含量、尿液尿素氮含量以及肾脏TGF-β、Collagen-1、NLRP3、IL-1β、TNF-α的转录水平明显降低(P<0.05,P<0.01),IL-10转录水平明显升高(P<0.01);与粪肠球菌组比较,粪肠球菌+绿原酸组的24 h尿钠水平有上升趋势但差异无统计学意义,粪肠球菌+绿原酸组血钠浓度和α-ENaC的转录及蛋白水平明显降低(P<0.01),粪肠球菌+绿原酸组的尿量、β-ENaC和γ-ENaC的转录水平则无明显变化(P>0.05)。体内和体外实验结果证明,绿原酸在一定程度上能抑制E.faecalis生长(P<0.01)。结论 绿原酸可改善粪肠球菌引起的高血压和肾损伤,其可能通过抑制E.faecalis生长和抑制α-ENaC通道活性及表达来改善钠潴留从而协同发挥降压作用。Objective To explore the antihypertensive and renal protective effects of chlorogenic acid(CGA)on Enterococcus faecalis(E.faecalis)-induced hypertension in mice.Methods Twenty C57BL/6 mice(6-8 weeks old)were randomly divided into control group,E.faecalis treated group,CGA treated group and E.faecalis+CGA treated group.10~9 CFU E.faecalis or 100 mg·kg^(-1)CGA was orally administered to mice for 8 weeks,during which blood pressure of mice was measured by tail cuff method.At the end of the experiment,24-hour urine samples were collected by metabolic cages.Serum and kidney samples were collected for the following examination.The concentration of urea nitrogen,sodium ion and urinary albumin were detected by kits;the albumin content of urine was detected by ELISA;the mRNA levels of TGF-β,Collagen-1,NLRP3,IL-1β,IL-10,TNF-α,epithelial sodium channel(ENaC)and the relative abundance of E.faecalis in feces were detected by RT-qPCR.The protein levels of α-ENaC were detected by Western Blot,and the optical density of E.faecalis in culture was detected by ultraviolet spectrophotometry.Results The results showed that,compared to control group,systolic blood pressure(SBP),urine albumin,urea nitrogen,renal injury factor mRNA,serum sodium and α-ENaC expression were significantly increased by E.faecalis treatment(P<0.05,P<0.01),while urinary sodium and the mRNA level of IL-10 were significantly decreased(P<0.05).Compared to E.faecalis treated group,SBP,24-hour urinary albumin,urinary urea nitrogen content,and mRNA levels of TGF-β,Collagen-1,NLRP3,IL-1β and TNF-αwere significantly decreased in E.faecalis+CGA treated group(P<0.05,P<0.01).The mRNA level of IL-10 was significantly increased(P<0.01).Furthermore,we found that 24-hour urinary sodium showed an upward trend after the treatment of E.faecalis+CGA,but there was no statistical difference.mRNA and protein levels of serum sodium,α-ENaC in E.faecalis+CGA treated group were significantly decreased(P<0.01),but urine volume,mRNA levels of β-ENaC and γ-ENaC did not signifi

关 键 词：绿原酸 粪肠球菌 高血压 肾损伤 上皮钠通道 小鼠 

分 类 号：R285.5[医药卫生—中药学]