标题：益气散聚方对肥胖相关肾小球病模型小鼠AMPK信号通路的影响及机制研究

      作者：方明珠 1，2，徐艳秋 1，顾耀东 1    1.上海中医药大学附属岳阳中西医结合医院肾内科，上海 200437；2.上海市第六人民医院金山分院中医内科，上海 201599

      卷次： 2022年33卷9期

      【摘要】 目的 探讨益气散聚方对肥胖相关性肾小球病模型小鼠体内AMPK与炎症诱发的关系以及对肾损害的影响，阐明益气散聚方防治肥胖相关性肾小球的作用机制。方法 随机将6周龄的60只雄性C57BL/6试验小鼠分为正常组(6只)、西药组(12只)、中药组(18只)、中西医结合组(12只)、模型组(12只)，正常组小鼠用生理盐水与普通饲料灌胃、西药组小鼠用二甲双胍与高脂饲料灌胃、中药组小鼠用益气散聚方与高脂饲料灌胃、中西医结合组小鼠用二甲双胍、益气散聚方与高脂饲料灌胃、模型组小鼠用生理盐水与高脂饲料灌胃。在药物干预的第4周末、第8周末观察各组小鼠的肥胖体质量，并检测24 h尿蛋白定量、尿RBP、尿NAG的变化情况，利用HE染色、免疫组化于镜下观察肾脏组织的病理变化，同时对肾脏组织内转化生长因子β1蛋白(TGF-β1)、单核细胞趋化蛋白-1(MCP-1)、磷酸化腺苷酸活化蛋白激酶α (P-AMPKα) mRNA含量进行检测。结果 在用药的第4周末和第8周末，正常组、中药组、模型组、西药组的小鼠体质量较前一次测量均增大，但差异无统计学意义(P>0.05)；中西医结合组小鼠与其他各组相比体质量下降明显，其差异均具有统计学意义(P<0.05)；在用药的第4周末和第8周末，中药组、中西医结合组、西药组与模型组相比，小鼠的24 h尿蛋白定量、尿RBP、尿NAG含量均降低，差异均具有显著统计学意义(P<0.01)，且五组进行组间比较发现，中西医结合组与中药组的治疗效果较好，差异具有统计学意义(P<0.05)；模型组、中药组、西药组、中西医结合组小鼠治疗的第4周末肾小球体积增大，肾小球囊腔变小，固有细胞无增生，系膜区未见明显增生，肾小管间质正常，未见细胞浸润；治疗第8周末检查发现与西药组、模型组进行比较，中西医结合组与中药组小鼠的肾小球直径均减小，但差异无统计学意义(P>0.05)；药物干预第8周末对五组小鼠免疫组化的结果进行比较发现，中西医结合组、西药组、中药组比模型组的PAMPK-α含量升高明显，差异具有显著统计学意义(P<0.01)；实时定量PCR检测结果显示：中药组、中西医结合组小鼠治疗的第 4周末、第 8周末MCP-1 mRNA、TGF-β1 mRNA表达下降，两组较模型组明显降低，差异均具有显著统计学意义(P<0.01)；中药组、西药组、中西医结合组小鼠用药的第 8周末PAMPK-α mRNA表达升高，三组较模型组显著升高，差异均具有统计学意义(P<0.05)。结论 益气散聚方可通过提高肥胖相关肾小球病模型小鼠AMPK表达，抑制MCP-1、TGF-β1等炎症因子表达，从而改善肥胖引起的肾脏损伤。
      【关键词】 C57BL/6试验小鼠；肥胖相关性肾病；益气散聚方；AMPK信号通路；炎症因子
      【中图分类号】 R-332 【文献标识码】 A 【文章编号】 1003—6350（2022）09—1089—07

Effect of Yiqi Sanju Recipe on AMPK signaling pathway of obesity-related glomerulopathy model mice and itsmechanism.
FANG Ming-zhu 1,2, XU Yan-qiu 1, GU Yao-dong 1. 1. Yueyang Hospital of Integrated Traditional Chinese andWestern Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 200437, CHINA; 2. Department ofTraditional Chinese Medicine, Jinshan Branch, Shanghai Sixth People's Hospital, Shanghai 201599, CHINA
【Abstract】 Objective To investigate the effect of Yiqi Sanju Recipe on AMPK signaling pathway, inflamma-tion, renal damage in obesity-related glomerulopathy model mice, and to clarify the mechanism of Yiqi Sanju Recipe inthe prevention and treatment of obesity-related glomerulopathy. Methods Sixty male C57BL/6 test mice at 6 weeks ofage were randomly divided into normal group (n=6); western medicine group (n=12); traditional Chinese medicinegroup (n=18); integrated traditional Chinese and western medicine group (n=12); model group (n=12); which were ga-vaged with normal saline and common feed; metformin and high-fat feed; Yiqi Sanju Recipe and high-fat feed; metfor-min, Yiqi Sanju Recipe, and high-fat feed; normal saline and high-fat feed; respectively. At the end of the 4th and 8th weekof treatment, the obesity body mass of mice in each group was observed, and the changes of 24 h urine protein, urine reti-nol-binding protein (RBP), and urine NAG were detected. The pathological changes of renal tissues were observed by HEstaining and immunohistochemistry under microscope. Meanwhile, the mRNA levels of transforming growth factor β 1(TGF-β1), monocyte chemotaxis protein-1 (McP-1), and phosphorylated adenylate activated protein kinase α (P-AMPKα)