当归多糖对糖尿病肾病KK-Ay小鼠肾脏AMPK信号通路及线粒体自噬的影响  

Effect of Angelica sinensis polysaccharides on AMPK signaling pathway and mitochondrial autophagy in kidney of diabetic nephropathy KK-Ay mice

作　　者：王江侠 杨丽霞[2] 米登海 魏瑞贤 崔阳阳 马仙康 WANG Jiang-xia;YANG Li-xia;MI Deng-hai;WEI Rui-xian;CUI Yang-yang;MA Xian-kang(Gansu University of Traditional Chinese Medicine,Lanzhou 730000,China;Gansu Province Academy of Chinese Medicine,Lanzhou 730050,China)

机构地区：[1]甘肃中医药大学,甘肃兰州730000 [2]甘肃省中医药研究院,甘肃兰州730050

出　　处：《中草药》2023年第10期3189-3196,共8页Chinese Traditional and Herbal Drugs

基　　金：陇药大品种二次开发及临床疗效评价行业技术中心(2019);甘肃省省属科研院所条件建设专项(20JR10RA432)。

摘　　要：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(AMPactivated 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,m TOR)蛋白表达;采用qRT-PCR检测肾组织AMPK、m TOR 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m RNA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 KKAy 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 t

关 键 词：当归多糖 糖尿病肾病 KK-AY小鼠 线粒体自噬 磷酸腺苷激活的蛋白激酶信号通路 

分 类 号：R285.5[医药卫生—中药学]