黄葵素抑制巨噬细胞浸润和活化改善db/db小鼠肾纤维化实验研究  

Experimental study of total flavones of Abelmoschus manihot to ameliorate renal fibrosis of db/db mice on the inhibition of infiltration and activation of macrophage

作　　者：浦强 徐巍龙[1] 李楠[1] 王丽娟 余江毅[1] PU Qiang;XU Wei-long;LI Nan;WANG Li-juan;YU Jiang-yi(Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210029,China;Rugao Hospital of Chinese Medicine,Rugao 226500,China)

机构地区：[1]南京中医药大学附属医院,南京210029 [2]如皋市中医院,如皋226500

出　　处：《天然产物研究与开发》2019年第11期1887-1895,共9页Natural Product Research and Development

基　　金：国家自然科学基金(8177141574);江苏省中医管理局重点专项(ZX2016A1);江苏省研究生培养创新工程研究生科研和实践创新计划(KYCX19_1207,KYCX18_1574);江苏省中医院2019院级课题“益中”助研基金(Y19041)

摘　　要：为探讨黄葵素对糖尿病肾病肾纤维化治疗作用及机制,采用SPF级C57BLKS/J db/db雄性小鼠20只,随机分为黄葵素低、中、高剂量(97.5、195、390 mg/kg/d)给药组和模型组4组,以同背景的5只db/m小鼠为空白对照组,分别用黄葵素混悬液及等体积的0.5%羧甲基纤维素钠溶液灌胃,连续16周,第24周处死,收集血和肾脏。检测尿ACR、血肌酐、血尿素氮,HE和Masson染色观察肾脏形态学改变和纤维化程度,免疫组化染色检测肾组织巨噬细胞标记F4/80表达情况,Western blot法检测肾组织F4/80蛋白、胶原蛋白Ⅲ、波形蛋白表达情况,qPCR法检测肾组织TNF-α、IL-1β和iNOS的mRNA水平。结果显示:与模型组相比,黄葵素可以降低db/db小鼠尿ACR、血肌酐、血尿素氮水平,减轻肾脏病理损伤及胶原纤维的沉积,降低肾组织胶原蛋白Ⅲ、波形蛋白及F4/80蛋白表达,减少肾组织F4/80标记巨噬细胞浸润;并下调了肾组织TNF-α、IL-1β、iNOS的mRNA水平。综上表明黄葵素可有效缓解db/db小鼠肾脏炎症反应,改善肾功能及肾纤维化,发挥肾保护作用,其机制与减少肾组织内巨噬细胞浸润及抑制促炎的M1型巨噬细胞极化相关。To explore the therapeutic effect and mechanism of total flavones of Abelmoschus manihot(TFA)on renal fibrosis in diabetic kidney disease,20 male SPF-C57BLKS/J db/db mice were randomly divided into 4 groups,namely TFA low-dose group(97.5 mg/kg/d),TFA middle-dose group(195 mg/kg/d),TFA high-dose group(390 mg/kg/d)and the model group.Five db/m mice in the same background were used as the blank control group.Mice were intragastrically administered with TFA’s suspension or an equal volume of 0.5%sodium carboxymethylcellulose solution for 16 weeks.At the 24th week mice were sacrificed and the blood and kidneys were collected.Urinary ACR,blood serum creatinine,and blood urea nitrogen were detected.HE staining and Masson staining were used to observe the changes of renal morphology and renal fibrosis.The expression of F4/80 in renal tissue macrophage was detected by immunohistochemical staining.The F4/80,collagen Ⅲ and vimentin proteins in renal tissue were detected by Western Blot.The expressions of TNF-α,IL-1βand iNOS were detected by qPCR method.The results showed that:compared with the model group,TFA could reduce urinary ACR,serum creatinine,blood urea nitrogen,lighten the pathological damage and collagen fiber deposition of the kidney,decrease the expression of collagen Ⅲ,vimentin and F4/80 proteins in mice renal tissues,reduce F4/80 labeled macrophage infiltration in renal tissue,and down-regulate the mRNA expressions of TNF-α,IL-1βand iNOS in renal tissues.In summary,TFA can effectively alleviate inflammatory response in db/db mice,improve renal function and renal fibrosis,and play a role in renal protection.Its mechanism is related to inhibition of macrophage infiltration in renal tissues and pro-inflammatory M1 macrophage polarization.

关 键 词：黄葵素 肾纤维化 巨噬细胞 炎症 DB/DB小鼠 

分 类 号：R285.5[医药卫生—中药学]