【摘要】 目的 探讨益气活血固肾颗粒对糖尿病肾病（DN）模型小鼠的治疗作用及其抑制炎症和氧化应激的相关机制。方法 将40只C57BL/6雄性小鼠随机分为正常组、模型组、益气活血固肾颗粒组(585 mg·kg^-1)、二甲双胍组(100 mg·kg^-1)。采用高脂饲料喂养结合腹腔注射链脲佐菌素（STZ）建立糖尿病肾病小鼠模型。造模成功后，按上述剂量给药，每日1次，连续10周。测量小鼠体质量、血糖、血脂等糖脂代谢水平及脏器系数、血清肌酐、尿素氮、蛋白尿等肾功能指标；采用Masson、PAS、HE染色观察小鼠肾脏病理变化；采用ELISA法检测小鼠炎症因子白细胞介素1β（IL-1β）、白细胞介素6（IL-6）、肿瘤坏死因子α（TNF-α）水平，用试剂盒测定氧化应激指标过氧化氢酶（CAT）、丙二醛（MDA）、超氧化物歧化酶（SOD）、谷胱甘肽（GSH）水平；采用RTPCR及Western Blot法检测肾组织中纤维化指标肌动蛋白（α-SMA）、转化生长因子β2（TGF-β2）、E-钙粘着蛋白（E-cadherin），以及炎症通路指标炎性小体（NLRP3）、1L-1β、炎性半胱氨酸蛋白酶（Caspase-1）和氧化应激通路指标Kelch样ECH关联蛋白1（Keap1）、核因子E2相关因子2（Nrf2）及炎症和氧化关联靶点NADPH氧化酶4（Nox4）的基因和蛋白表达。结果 与正常组比较，模型组小鼠体质量、血糖、肝脏系数、肾脏系数、血尿素氮（BUN），尿蛋白（UAE）、血肌酐（Scr）、尿蛋白/尿肌酐（ACR）、甘油三酯（TG）、总胆固醇（TC）、低密度脂蛋白（LDL-C）、MDA、IL-1β、IL-6、TNF-α水平显著上升（P<0.001），脾脏系数、尿肌酐（Ucr）、高密度脂蛋白（HDL-C）、CAT、SOD、GSH水平明显降低（P<0.001），肾脏中α-SMA、TGF-β2、NLRP3、Caspase-1、1L-1β、 Keap1、 Nox4 mRNA及蛋白表达水平显著上升（P<0.01, P<0.001），肾脏中E-cadherin和Nrf2mRNA及蛋白表达水平显著降低（P<0.05,P<0.001）。与模型组比较，益气活血固肾颗粒组和二甲双胍组小鼠体质量、血糖、肝脏系数、肾脏系数、BUN、UAE、Scr、ACR、TG、TC、LDL-C、MDA、IL-1β、IL-6、TNF-α水平显著下降（P<0.05,P<0.01,P<0.001），脾脏系数、Ucr、HDL-C、CAT、SOD、GSH水平明显上升（P<0.05,P<0.01,P<0.001），肾脏中α-SMA、TGF-β2、NLRP3、Caspase-1、1L-1β、Keap1、Nox4的mRNA及蛋白表达水平显著下降（P<0.05,P<0.01,P<0.001），肾脏中E-cadherin和Nrf2的mRNA及蛋白表达水平显著升高（P<0.05,P<0.01,P<0.001）。结论 益气活血固肾颗粒可以通过调控Nox4介导的NLRP3/Caspase-1和Nrf2/Keap1信号通路抑制炎症和氧化应激，改善肾脏纤维化，从而达到治疗糖尿病肾病的效果。更多还原

【Abstract】 Objective To investigate the therapeutic effects of Yiqi Huoxue Gushen Granules（YHGG） on diabetic nephropathy（DN） model mice and its mechanisms related to inhibiting inflammation and oxidative stress. Methods Forty male C57BL/6 mice were randomly divided into a normal group,a model group,a YHGG group(585 mg·kg^-1),and a metformin group(100 mg·kg^-1). A DN mouse model was established by feeding a high-fat diet combined with intraperitoneal injection of Streptozotocin（STZ）. After successful modeling, the mice were administered the corresponding doses once daily for 10 weeks. Body mass, blood glucose, lipid metabolism levels, organ indices,serum creatinine, blood urea nitrogen（BUN）, and proteinuria were measured. Renal pathological changes were observed using Masson, PAS, and HE staining. Serum levels of inflammatory factors interleukin-1β（IL-1β）,interleukin-6（IL-6）,and tumor necrosis factor-α（TNF-α） were measured by ELISA. Oxidative stress indicators,including catalase（CAT）, malondialdehyde（MDA）, superoxide dismutase（SOD）, and glutathione（GSH）,were determined using assay kits. RT-PCR and Western Blot were used to detect the mRNA and protein expression of fibrosis markers（α-SMA,TGF-β2,E-cadherin）,inflammatory pathway markers（NLRP3,IL-1β,Caspase-1）,oxidative stress pathway markers（Keap1,Nrf2）,and the inflammation and oxidative stress-related target NADPH oxidase 4（Nox4） in renal tissues. Results Compared with the normal group,the model group showed significantly increased body mass,blood glucose,liver index,kidney index,BUN,urinary albumin excretion（UAE）,serum creatinine（Scr）,albumin-to-creatinine ratio（ACR）,triglycerides（TG）,total cholesterol（TC）,low-density lipoprotein cholesterol（LDL-C）,MDA,IL-1β,IL-6,and TNF-α levels（P<0.01）,and significantly decreased spleen index, urinary creatinine（Ucr）, high-density lipoprotein cholesterol（HDL-C）, CAT, SOD, and GSH levels（P<0.01）. The mRNA and protein expressions of α-SMA,TGF-β2,NLRP3,Caspase-1,IL-1β,Keap1,and Nox4 in renal tissues was significantly increased（P<0.01,P<0.001）,while the mRNA and protein expression of E-cadherin and Nrf2 was significantly decreased（P<0.05, P<0.001）. Compared with the model group, the YHGG group and metformin group showed significantly decreased body mass, blood glucose, liver index, kidney index,BUN,UAE,Scr,ACR,TG,TC,LDL-C,MDA,IL-1β,IL-6,and TNF-α levels（P<0.05,P<0.01, P<0.001）, and significantly increased spleen index, Ucr, HDL-C, CAT, SOD, and GSH levels（P<0.05, P<0.01, P<0.001）. The mRNA and protein expression of α-SMA, TGF-β2, NLRP3, Caspase-1,IL-1β,Keap1,and Nox4 in renal tissues was significantly decreased（P<0.05,P<0.01,P<0.001）,while the mRNA and protein expression of E-cadherin and Nrf2 was significantly increased（P<0.05,P<0.01,P<0.001）. Conclusion YHGG can inhibit inflammation and oxidative stress by regulating the Nox4-mediated NLRP3/Caspase-1 and Nrf2/Keap1 signaling pathways, thereby improving renal fibrosis and achieving therapeutic effects in diabetic nephropathy.更多还原