目的 研究当归多糖对糖尿病肾病（diabetic nephropathy，DN）KK-Ay小鼠肾脏磷酸腺苷激活的蛋白激酶（AMP-activated protein kinase，AMPK）信号通路及线粒体自噬的影响。方法 SPF级雄性KK-Ay小鼠用高糖高脂饲料喂养，随机分为模型组、厄贝沙坦（25 mg/kg）组和当归多糖高、中、低剂量（400、200、100 mg/kg）组，每组10只；将10只雄性C57BL/6J小鼠作为对照组。给予药物干预4周，观察小鼠一般情况，每周称定体质量并检测血糖；末次给药后，心脏取血并处死小鼠，分离血清检测尿微量白蛋白（urine microalbuminuria，U-ALB）、肌酐（creatinine，SCr）、尿素氮（urea nitrogen，BUN）；采用苏木素-伊红（HE）染色观察肾组织病理变化；采用Western blotting检测肾组织线粒体自噬相关蛋白[微管相关蛋白1轻链3（microtubule-associated protein 1 light chain 3，LC3）、p62、Nix]和线粒体裂变蛋白[线粒体动力相关蛋白1（dynamin-related protein 1，Drp1）]的表达；采用免疫组化法检测肾组织AMPK、哺乳动物雷帕霉素靶蛋白（mammalian target of rapamycin，mTOR）蛋白表达；采用qRT-PCR检测肾组织AMPK、mTOR mRNA表达。结果 与模型组比较，各给药组U-ALB、SCr、BUN水平均明显降低（P＜0.05、0.01），呈剂量相关性；肾组织病理变化有所改善；肾组织线粒体自噬相关蛋白LC3Ⅱ/LC3Ⅰ、Nix蛋白表达水平显著降低（P＜0.01），p62蛋白表达水平显著升高（P＜0.05）；线粒体裂变蛋白Drp1表达下调（P＜0.01）；AMPK、mTOR蛋白及mRNA表达显著下调（P＜0.05、0.01）。结论 当归多糖能改善DN小鼠肾损伤，延缓DN发病进展，其作用机制与抑制AMPK信号通路介导的线粒体自噬有关。

Objective To study the effects of Angelica sinensis polysaccharides (ASP) on AMP activated protein kinase (AMPK) signaling pathway and mitochondrial autophagy in kidney of KK-Ay mice with diabetes nephropathy (DN). Methods SPF male KK-Ay mice were fed with high sugar and high fat diet and randomly divided into model group, irbesartan (25 mg/kg) group, ASP high-, medium-, and low-dose (400, 200, 100 mg/kg) groups, with 10 mice in each group, 10 male C57BL/6J mice were used as control group. Drugs were given for intervention for four weeks, the general condition of mice was observed, body weight and blood sugar were measured weekly; After the last administration, blood was taken from heart and mice were euthanized. Serum was separated and tested for urinary microalbuminuria (U-ALB), creatinine (SCr), and urea nitrogen (BUN); Pathological changes of renal tissue was observed by hematoxylin eosin (HE) staining; Western blotting was used to detect the expressions of mitochondrial autophagy related proteins [microtubule associated protein 1 light chain 3 (LC3), p62, Nix] and mitochondrial fission protein [mitochondrial related protein 1 (Drp1)] in renal tissue; Immunohistochemical method was used to detect the expressions of AMPK and mammalian target of rapamycin (mTOR) protein in renal tissue; The expressions of AMPK and mTOR mRNA in renal tissue were detected by qRT-PCR. Results Compared with model group, levels of U-ALB, SCr and BUN in each treatment group were significantly reduced (P < 0.05, 0.01), showing a dose-dependent relationship; The pathological changes of renal tissue was improved; The expression levels of mitochondrial autophagy related proteins LC3II/LC3I and Nix in renal tissue were significantly reduced (P < 0.01), while the expression level of p62 protein was significantly increased (P < 0.05); The expression of mitochondrial fission protein Drp1 was downregulated (P < 0.01); AMPK, mTOR protein and mRNA expressions were significantly downregulated (P < 0.05, 0.01). Conclusion ASP can improve renal injury in DN mice and delay the progression of DN. Its mechanism is related to the inhibition of AMPK signaling pathway mediated mitochondrial autophagy.

R285.5

2021年国家中医药管理局青年岐黄学者项目（国中医药人教函[2022]6号）