基于“肠-肾轴”理论探讨芪黄四物汤对环孢素A慢性肾毒性的保护机制  

Protective mechanism of Qihuang Siwu Decoction on cyclosporine A chronic nephrotoxicity based on the theory of'gut kidney axis'

作　　者：韩聪 胡洪贞[2] 周乐[2] 姜月华[2] 刘瑶[3] 杨旭[3] 吕帆[3] 宋国栋 李伟[2] HAN Cong;HU Hong-zhen;ZHOU Le;JIANG Yue-hua;LIU Yao;YANG Xu;LYU Fan;SONG Guo-dong;LI Wei(First Clinical School of Medicine,Shandong University of Traditional Chinese Medicine,Jinan 250014,China;Affiliated Hospital of Shandong University of TCM,Jinan 250014,China;Shandong University of Traditional Chinese Medicine,Jinan 250014,China)

机构地区：[1]山东中医药大学第一临床医学院,济南250014 [2]山东中医药大学附属医院,济南250014 [3]山东中医药大学,济南250014

出　　处：《中华中医药杂志》2020年第5期2623-2627,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金面上项目(No.81673812);山东省重点研发计划项目(No.2018GSF119017)。

摘　　要：目的:探讨芪黄四物汤通过"肠-肾轴"调节肠道菌群进而对环孢素A(CsA)慢性肾毒性的保护作用。方法:24只雄性C57BL/6小鼠随机分为3组:对照组、模型组及芪黄四物汤组,每组8只,以CsA 30mg·kg-1·d-1灌胃建立CsA慢性肾毒性模型,造模时芪黄四物汤组给予0.48g·kg-1·d-1灌胃,模型组及对照组等量0.9%氯化钠溶液灌胃,6周后观察3组小鼠一般情况及肾功能、炎症及尿毒素指标[血肌酐(Scr)、尿素氮(BUN)、24h尿蛋白定量、白介素-6(IL-6)、硫酸吲哚酚(IS)],HE、Masson染色观察结肠组织及肾脏病理,免疫荧光观察结肠组织闭锁小带蛋白1(ZO-1)及肾组织转化生长因子β1(TGF-β1)表达,并提取小鼠肠道微生物总DNA进行16S rDNA V3+V4区测序。结果:与对照组比较,模型组小鼠体质量降低,尿量减少,Scr、BUN、24h尿蛋白定量、IL-6、IS指标均显著升高(P<0.05);与模型组比较,芪黄四物汤干预后均显著改善(P<0.05)。与对照组比较,模型组小鼠结肠组织ZO-1表达减少,肾组织示肾小球基底膜增厚、小动脉玻璃样变,肾小管空泡变性伴有萎缩、间质炎症细胞浸润、增宽伴有条带状纤维化,胶原纤维及TGF-β1表达增加;芪黄四物汤干预后均有不同程度改善。与对照组比较,模型组小鼠菌群丰度与多样性均降低,厚壁菌门与拟杆菌门比值(F/B)升高(P<0.01),拟杆菌属等致病菌含量增加(P<0.05),乳杆菌属等益生菌含量降低;芪黄四物汤增加了阿克曼属等益生菌含量,改善了菌群紊乱。结论:芪黄四物汤能够通过"肠-肾轴"降低CsA慢性肾毒性小鼠肠道通透性,增加阿克曼属等益生菌的含量、降低拟杆菌属等致病菌的含量,改善炎症及纤维化状态,降低尿毒素水平,保护肾功能。Objective: To explore the protective effect of Qihuang Siwu Decoction on cyclosporine A(CsA) chronic nephrotoxicity by regulating intestinal flora through ’gut kidney axis’. Methods: Twenty-four male C57 BL/6 mice were randomly divided into three groups: control(W) group, model(M) group and Qihuang Siwu Decoction(QHSW) group, 8 mice in each group. CsA chronic nephrotoxicity model was established by gavage of CsA 30 mg·kg-1·d-1. The QHSW group was given 0.48 g·kg-1·d-1 QHSW by gavage, the M group and the W group were given 0.9% sodium chloride solution by gavage. Six weeks later, the general situation and kidney function, inflammation and urotoxin(Scr, BUN, 24 pro, IL-6, IS) of the three groups were observed. HE and Masson staining were used to observe the pathological changes of colon and kidney. Immunofluorescence was used to observe the expression of ZO-1 and TGF-β1 in colon and kidney. The total DNA of mouse intestinal microorganism was extracted and sequenced in V3+V4 region of 16 S rDNA. Results: Compared with the W group, the body weight and urine volume of the M group decreased, and the indexes of Scr, BUN, 24 pro, IL-6 and IS increased(P<0.05). The prognosis of QHSW group was significantly improved compared with the M group(P<0.05). Compared with the W group, the expression of ZO-1 in colon decreased in M group, glomerular basement membrane thickening, renal tubule expansion, inflammatory cell infiltration, collagen fiber and TGF-β1 increased in renal tissue, and the prognosis of QHSW group was improved in varying degrees. Compared with the W group, the abundance and diversity of flora in M group decreased, the ratio of Firmicutes to Bacteroides(F/B) in M group increased(P<0.01), the content of Bacteroides and other pathogens increased(P<0.05), the content of Lactobacillus and other probiotics decreased, QHSW increased the content of Akerman and other probiotics, and improved the flora disorder. Conclusion: QHSW can reduce the intestinal permeability, increase the content of probiotics such as A

关 键 词：16SrDNA测序 芪黄四物汤 肠道菌群 慢性肾脏病 环孢素A慢性肾毒性 机制 “肠-肾轴”理论 

分 类 号：R285.5[医药卫生—中药学]