补阳还五汤加味通过抑制炎症和纤维化改善糖尿病肾病小鼠肾脏损伤  

Supplemented Buyang Huanwutang Ameliorates Kidney Injury by Inhibiting Inflammation and Fibrosis in Diabetic Kidney Disease Mice

作　　者：杨帆[1] 张晓云 张亚京 刘利飞 黄家安 王月华 YANG Fan;ZHANG Xiaoyun;ZHANG Yajing;LIU Lifei;HUANG Jiaan;WANG Yuehua(Hebei University of Chinese Medicine,Shijiazhuang 050091,China)

机构地区：[1]河北中医学院,石家庄050091

出　　处：《中国实验方剂学杂志》2022年第24期114-121,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：河北中医学院博士科研基金项目(BSZ2022003);河北省自然科学基金项目(H2022423320);河北省中医药管理局科研计划项目(2023119)。

摘　　要：目的:探讨补阳还五汤加味对糖尿病肾病(DKD)小鼠肾组织保护作用及核转录因子-κB(NF-κB)通路及纤维化因子的影响。方法:24只11~12周龄db/db小鼠适应性喂养1周并监测尿蛋白均阳性后随机分为模型组、补阳还五汤加味组(16.0 g·kg^(-1))、厄贝沙坦组(13.5 mg·kg^(-1)),每组8只。11~12周龄db/m组小鼠8只作为正常组。给予相应药物治疗,其中补阳还五汤加味组及厄贝沙坦组灌胃相应药液,正常组及模型组灌胃等体积蒸馏水,每天1次,连续治疗8周后收集标本,检测小鼠血糖(FBG)、总胆固醇(TC)、甘油三脂(TG)、尿素氮(BUN)、肌酐(SCr)、尿微量白蛋白(mALB)水平;实时荧光定量聚合酶链式反应(Real-time PCR)检测肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)和单核细胞趋化因子-1(MCP-1)的mRNA表达;蛋白免疫印迹法(Western blot)检测小鼠肾组织核转录因子-κB(NF-κB),NF-κB抑制因子α(IκBα)、磷酸化IκBα(p-IκBα)、转化生长因子-β_(1)(TGF-β_(1))、α-平滑肌肌动蛋白(α-SMA)、纤连蛋白(FN)的蛋白表达;免疫荧光检测肾组织NF-κB表达;同时光镜观察肾脏病理形态学变化。结果:与正常组比较,模型组小鼠肾小球肥大,系膜外基质增加,基底膜增厚,囊腔变窄,间质炎性细胞浸润,部分间质明显纤维化(P<0.01);FBG、mALB、TC、TG、BUN、SCr含量显著升高(P<0.01);炎症因子TNF-α、IL-1β、MCP-1及纤维化相关蛋白TGF-β_(1)、α-SMA、FN表达显著升高(P<0.01);同时肾组织NF-κB通路活化程度显著增强(P<0.01)。与模型组比较,补阳还五汤加味干预后,DKD小鼠肾脏病理损伤程度显著改善(P<0.01);mALB、TC、TG含量显著下降,BUN、SCr含量显著降低(P<0.01);TNF-α、IL-1β、MCP-1含量明显降低(P<0.05,P<0.01);同时抑制了肾组织NF-κB通路活化及纤维化因子表达(P<0.05,P<0.01),但是对血糖水平无明显影响。结论:补阳还五汤加味可通过抑制NF-κB通路活化及纤维化因子表达,对Objective:To investigate the effect of supplemented Buyang Huanwutang on kidney tissue,nuclear factor-κB(NF-κB)pathway,and fibrosis factors in diabetic kidney disease(DKD)mice.Method:A total of 24 db/db mice(11-12 weeks old)were randomized into the model group(equivalent volume of distilled water,once/day,8 weeks),supplemented Buyang Huanwutang group(16.0 g·kg^(-1),once/day,8 weeks),and irbesartan group(13.5 mg·kg^(-1),once/day,8 weeks)after adaptive feeding for 1 week and positive urinary protein monitoring,with 8 in each group.Another 8 db/m mice(11-12 weeks old)were included in the normal group(equivalent volume of distilled water,once/day,8 weeks).Then samples were collected,and the levels of fasting blood glucose(FBG),total cholesterol(TC),triglyceride(TG),blood urea nitrogen(BUN),serum creatinine(SCr),and urinary microalbumin(mALB)were detected.The mRNA expression of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),and monocyte chemoattractant protein-1(MCP-1)was determined by real-time polymerase chain reaction(Real-time PCR).The expression of nuclear factor-κB(NF-κB),NF-κB inhibitorα(IκBα),phosphorylated IκBα(p-IκBα),transforming growth factor-β_(1)(TGF-β_(1)),α-smooth muscle actin(α-SMA),and fibronectin(FN)in kidney tissue was measured by Western blot.The expression of NF-κB in renal tissue was detected by immunofluorescence.The pathological changes of kidney were observed under light microscope.Result:Compared with the normal group,the model group showed glomerular hypertrophy,increase in extracellular matrix,thickening of basement membrane,small cystic lumen,interstitial inflammatory cell infiltration,and some interstitial fibrosis(P<0.01).Moreover,the model group had higher content of FBG,mALB,TC,TG,BUN,and SCr(P<0.01),higher expression of inflammatory factors TNF-α,IL-1β,and MCP-1,and fibrosis-related proteins TGF-β_(1),α-SMA,and FN(P<0.01),and stronger activation of NF-κB pathway in renal tissue(P<0.01)than the normal group.Compared with the model group,supplemented B

关 键 词：糖尿病肾脏疾病 补阳还五汤加味 核转录因子-κB(NF-κB)通路 炎症 纤维化 

分 类 号：R2-0[医药卫生—中医学] R33R289R587.1