槲皮素对小鼠单侧输尿管结扎致肾间质纤维化的改善作用及机制研究  

Improvement effect and mechanism of quercetin on renal interstitial fibrosis induced by unilateral ureteral ligation in mice

作　　者：黎艳 刘莉 王琪文 杨越 张静 刘丹 LI Yan;LIU Li;WANG Qiwen;YANG Yue;ZHANG Jing;LIU Dan(Jingmen People’s Hospital,Jingmen 448000,China)

机构地区：[1]荆门市人民医院荆楚理工学院附属中心医院,湖北荆门448000 [2]西安市第五医院,陕西西安710082 [3]上海市浦东医院复旦大学附属浦东医院风湿免疫科,上海201399

出　　处：《陕西医学杂志》2023年第11期1473-1477,1482,共6页Shaanxi Medical Journal

基　　金：陕西省自然科学基础研究计划项目(2021JZ-59);湖北省荆门市科技计划项目(2023YDKY054)。

摘　　要：目的:探讨槲皮素对小鼠单侧输尿管结扎(UUO)致肾间质纤维化的改善作用及机制。方法:将48只C57/B6健康雄性小鼠随机分为四组:对照组行单侧输尿管结扎术后给予皮下注射0.9%氯化钠溶液;低剂量给药组行单侧输尿管结扎术后给予皮下注射槲皮素3 mg/(kg·d);中剂量给药组行单侧输尿管结扎术后给予皮下注射槲皮素9 mg/(kg·d);高剂量给药组行单侧输尿管结扎术后给予皮下注射槲皮素18 mg/(kg·d)。分别于术后第14天处死小鼠,取输尿管结扎侧肾组织。检测小鼠血清肌酐和尿素氮含量;肾脏组织行PAS检查损伤程度,天狼星红染色检测肾间质纤维化程度;用RT-qPCR法检测肾脏组织中肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)、胶原蛋白1 (COL-1)、纤连蛋白(FN)、α-平滑肌肌动蛋白(α-SMA) mRNA相对表达量,用免疫组化检测肾组织中巨噬细胞变化,Western blot检测小鼠肾组织α-SMA、转化生长因子β1(TGF-β1)蛋白表达。结果:PAS染色和天狼星红染色结果表明,单侧输尿管结扎术后肾脏间质区胞外基质成分增多细胞成分减少,肾小管萎缩而肾小球病变不明显,肾组织出现间质纤维化改变。槲皮素治疗后,低剂量和中剂量给药组血清肌酐及尿素氮水平相比对照组无统计学差异(均P>0.05),高剂量组血清肌酐及尿素氮相比对照组降低(均P<0.05);高剂量组与对照组相比较炎症因子TNF-α、IL-6和纤维化因子COL-1、FN、α-SMA mRNA相对表达量下调;Western blot结果显示,槲皮素高剂量组抑制UUO小鼠肾组织α-SMA、TGF-β1的表达。结论:槲皮素可以改善UUO小鼠肾间质纤维化指标以及血肌酐和尿素氮含量,其机制可能是通过降低炎症细胞浸润,并通过下调α-SMA和TGF-β1蛋白表达,改善肾小管上皮细胞间充质转化,从而减轻纤维化程度。Objective:To investigate the improvement effect and mechanism of quercetin on renal interstitial fibrosis induced by unilateral ureteral obstruction(UUO)in mice.Methods:Forty-eight healthy C57/B6 male mice were randomly divided into four groups:the control group was given subcutaneous injection of 0.9%sodium chloride solution after UUO;in the low-dose treatment group,subcutaneous injection of 3 mg/kg of quercetin was given after UUO;in the middle-dose group,subcutaneous injection of 9 mg/kg of quercetin was given after UUO;in the high-dose group,subcutaneous injection of 9 mg/kg of quercetin was given after UUO.The mice were sacrificed on the 14th day after operation,and the kidney tissues of the ligated side were collected.The levels of serum creatinine and urine urea nitrogen were detected.The degree of renal injury was examined by PAS,and the degree of renal interstitial fibrosis was detected by sirius red staining.The relative expression of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),collagen 1(COL-1),fibronectin(FN)andα-smooth muscle actin(α-SMA)mRNA in renal tissue was detected by RT-PCR,and the change of macrophages in renal tissue was detected by immunohistochemistry.The protein expressions ofα-SMA and transforming growth factor-β1(TGF-β1)in renal tissue were detected by Western blot.Results:The results of PAS and sirius red staining showed that the extracellular matrix components increased and the cellular components decreased in the renal interstitium after UUO,the renal tubules atrophy without obvious glomerular lesions,and the interstitial fibrosis changes appeared in the renal tissue.After quercetin treatment,the levels of serum creatinine and urea nitrogen in the low-dose and medium-dose groups were not significantly different from those in the control group(all P>0.05),while the levels of serum creatinine and urea nitrogen in the high-dose group were lower than those in the control group(all P<0.05).Compared with the control group,the mRNA relative expression of inflammatory factors T

关 键 词：单侧输尿管结扎 肾间质纤维化 槲皮素 慢性肾脏病 炎症细胞浸润 间充质转化 

分 类 号：R915[医药卫生—微生物与生化药学]