基于自噬途径探讨补阳还五汤合抵挡汤改善尿酸性肾病大鼠肾脏损伤的作用机制  

Discussion on the mechanism of Buyang Huanwu Decoction and Didang Decoction improving renal injury in rats with uric acid nephropathy based on autophagy pathway

作　　者：范丽妃 郭玉琴 林敏[1] 翁曼华 鲁玉辉[1] FAN Lifei;GUO Yuqin;LIN Min;WENG Manhua;LU Yuhui(College of Traditional Chinese Medicine,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,China)

机构地区：[1]福建中医药大学中医学院,福州350122

出　　处：《中华中医药杂志》2024年第8期3989-3995,共7页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：福建省自然科学基金项目(No.2022J01360);福建中医药大学高层次人才科研启动基金项目(No.X2020007-人才),福建中医药大学基础与临床联合研究专项(No.X2023029)。

摘　　要：目的:探讨补阳还五汤合抵挡汤对尿酸性肾病(UAN)大鼠肾脏纤维化的治疗效果及相关作用机制。方法:48只雄性SPF级SD大鼠适应喂养1周后,随机分为空白对照组,模型组,别嘌醇组,补阳还五汤合抵挡汤高、中、低剂量组,每组8只。采用腺嘌呤(100 mg/kg)联合氧嗪酸钾(300 mg/kg)混悬液灌胃建立UAN大鼠模型,造模同时各给药组分别给予别嘌醇(5 mg/kg)和补阳还五汤合抵当汤高、中、低剂量(3.46、1.73、0.87 g/kg)灌胃,连续21 d。观察各组大鼠体质量、肾脏质量及肾脏指数等;检测血尿酸(SUA)、血肌酐(Scr)、尿素氮(BUN)及24 h尿微量白蛋白(24h mAlb)水平;HE染色观察肾组织病理变化,Masson染色观察肾脏胶原阳性面积占比情况,免疫组织化学染色检测肾组织CD68、α-平滑肌肌动蛋白(α-SMA)表达情况;实时荧光定量PCR检测肾组织自噬标志蛋白LC3-Ⅱ、Beclin-1、P62基因表达;Western Blot检测自噬标志蛋白LC3-Ⅱ/LC3-Ⅰ比值、Beclin-1、P62蛋白表达。结果:与空白对照组比较,模型组大鼠体质量显著降低(P<0.01),肾脏指数显著升高(P<0.01),血清SUA、Scr、BUN及尿液24h mAlb含量显著升高(P<0.01),肾脏胶原容积分数(CVF)显著升高(P<0.01),肾组织CD68、α-SMA阳性表达面积显著增加(P<0.01),肾组织自噬标志蛋白LC3-Ⅱ基因表达与LC3-Ⅱ/LC3-Ⅰ比值均显著升高(P<0.01),Beclin-1基因与蛋白表达显著升高(P<0.01),P62表达显著降低(P<0.01),并伴尿酸钠盐结晶沉积和肾组织结构损伤等病理改变。与模型组比较,补阳还五汤合抵当汤高、中剂量组大鼠体质量显著增加(P<0.01),别嘌醇组及补阳还五汤合抵当汤高、中剂量组肾脏指数显著降低(P<0.01),各给药组血清SUA、Scr、BUN水平显著降低(P<0.05,P<0.01),补阳还五汤合抵当汤高、中剂量组尿液24h mAlb含量显著降低(P<0.05,P<0.01),各药物干预组肾组织病理均有不同程度改善,以中剂量组更为接近空白对照组,别嘌醇�Objective:To investigate the therapeutic effect and mechanism of Buyang Huanwu Decoction and Didang Decoction on renal fibrosis in rats with uric acid nephropathy(UAN).Methods:A total of 48 male SPF SD rats were randomly divided into blank control group,model group,allopurinol group,Buyang Huanwu Decoction and Didang Decoction high,medium and low dose groups,with 8 rats in each group.The UAN model was established by intragastric administration of Adenine(100 mg/kg)combined with Potassium Oxonate(300 mg/kg).At the same time,Allopurinol(5 mg/kg)and Buyang Huanwu Decoction and Didang Decoction(3.46,1.73,0.87 g/kg)were given by intragastric administration for 21 days.Thebody weight,kidney weight and kidney index were observed.The levels of serum uric acid(SUA),serum creatinine(Scr),blood urea nitrogen(BUN)and 24h urinary microalbumin(24h mAlb)were detected.HE staining was used to observe thepathological changes of renal tissue.Masson staining was used to observe the proportion of collagen positive area in renaltissue.Immunohistochemical staining was used to detect the expressions of CD68 andα-SMA in renal tissue.The mRNAexpressions of autophagy marker proteins LC3-II,Beclin-1 and P62 in renal tissue were detected by real-time PCR.The ratioof autophagy marker protein LC3-II/LC3-I and the expression of Beclin-1 and P62 proteins were detected by Western Blot.Results:Compared with the blank control group,the body weight of rats in the model group significantly decreased(P<0.01),significantly increased renal index(P<0.01),serum SUA,Scr,BUN and urine 24 h mAlb content significantly increased(P<0.01),renal collagen volume fraction(CVF)significantly increased(P<0.01),and the positive expression areas of CD68 andα-SMAin renal tissues were significantly increased(P<0.01).The mRNA expression of autophagy marker protein LC3-II and the ratioof LC3-I/LC3-Iin renal tissue were significantly increased(P<0.01),and the mRNA and protein expression of Beclin-1 weresignificantly increased(P<0.01),and P62 expression was significantly de

关 键 词：补阳还五汤合抵挡汤 尿酸性肾病 肾脏损伤 自噬途径 机制 

分 类 号：R285.5[医药卫生—中药学]