摘要
目的:探讨五味子合剂(SM)对糖尿病肾病(diabetic nephropathy,DN)大鼠肾脏的保护作用,及其对自噬及Akt/mTOR通路的影响。方法:SPF级SD大鼠60只,随机分为对照组、模型组、达格列净(Dapagliflozin,0.9 g·kg^(-1)·d^(-1),DA)组、SM高剂量(6 g·kg^(-1)·d^(-1),SM-H)组、SM中剂量(3 g·kg^(-1)·d^(-1),SM-M)组及SM低剂量(1.5 g·kg^(-1)·d^(-1),SM-L)组,每组10只。除对照组外,其余各组均采用高糖高脂饮食联合STZ单次腹腔注射构建DN模型。造模成功后,干预12周。连续观测大鼠血糖(Glu)及24 h尿蛋白定量(24 h UTP)变化。干预结束后,检测肾功能,肾脏指数(KI),与肾组织病理变化。Western Blotting检测肾组织中自噬泡标记蛋白(LC3)、自噬底物(P62)、蛋白激酶B(Akt)、磷酸化Akt(p-Akt)、雷帕霉素靶蛋白(mTOR)以及磷酸化mTOR(p-mTOR)的蛋白表达量;免疫荧光染色检测肾组织中LC3、P62的表达情况。结果:与对照组相比,模型组Glu、24 h UTP、Scr、BUN及KI均显著升高(P<0.05或P<0.01),肾组织病理损伤明显加重;LC3Ⅱ/LC3Ⅰ蛋白表达量比值降低,P62蛋白表达量以及p-Akt/Akt、p-mTOR/mTOR表达量比值(P<0.05或P<0.01)升高。与模型组相比,SM各剂量组及DA组Glu、24 h UTP、Scr、BUN及KI均显著降低(P<0.05或P<0.01),肾组织病理损伤明显改善,SM-H组上述变化最为明显;并且SM-H组LC3Ⅱ/LC3Ⅰ蛋白表达量比值升高,P62蛋白表达量以及p-Akt/Akt、p-mTOR/mTOR表达量比值(P<0.05或P<0.01)降低。结论:SM可激活DN大鼠肾脏自噬,促进自噬降解,从而发挥肾保护作用,其机制可能与抑制Akt/mTOR通路有关。
Objective:To investigate the protective effect of Schisandra Chinensis Mixture(SM)on the kidney of diabetic nephropathy(DN)rats,and its effect on autophagy and Akt/mTOR pathway.Methods:Sixty:SPF SD rats were randomly divided into control group,model group,Dapagliflozin(0.9 g·kg^(-1)·d^(-1),DA)group,SM high dose(6 g·kg^(-1)·d^(-1),SM-H)group,SM medium dose(3 g·kg^(-1)·d^(-1),SM-M)group and SM low dose(1.5 g·kg^(-1)·d^(-1),SM-L)group,with 10 rats in each group.Except for the control group,the other groups were treated with high-sugar and high-fat diet combined with STZ single intraperitoneal injection to construct DN model.After successful modeling,the intervention lasted for 12 weeks.The changes of blood glucose(Glu)and 24-hour urinary protein(24 h UTP)were observed continuously.After the intervention,renal function,kidney index(KI),and pathological changes of renal tissue were detected.Western Blotting was used to detect the expression of autophagic vacuole marker protein(LC3),autophagic substrate(P62),protein kinase B(Akt),phosphorylated Akt(p-Akt),rapamycin target protein(mTOR)and phosphorylated mTOR(p-mTOR)in renal tissue.The expression of LC3 and P62 in renal tissue was detected by immunofluorescence staining.Results:Compared with the model group,Glu,24 h UTP,Scr,BUN and KI in all the SM dose groups and DA group were significantly decreased(P<0.05 or P<0.01),and the pathological damage of renal tissue was significantly improved,and the above changes were most obvious in the SM-H group;the ratio of LC3Ⅱ/LC3Ⅰprotein expression in SM-H group was increased,the expression of P62 protein and the ratio of p-Akt/Akt and p-mTOR/mTOR expression were decreased(P<0.05 or P<0.01).Conclusion:SM can activate autophagy in the kidney of DN rats,promote autophagy degradation,and thus play a role in renal protection.The mechanism may be related to the inhibition of Akt/mTOR pathway.
作者
张术姣
张帅星
张泽钰
白雪慧
郑慧娟
邓媛媛
张赛
马钰
刘伟敬
张勉之
ZHANG Shujiao;ZHANG Shuaixing;ZHANG Zeyu(Beijing University of Chinese Medicine Dongzhimen Hospital,Beijing,100700)
出处
《中国中西医结合肾病杂志》
2024年第12期1041-1044,I0009,I0010,共6页
Chinese Journal of Integrated Traditional and Western Nephrology
基金
国家自然科学基金资助项目(No.82074242,82074361,82374382)
天津市中医药研究院附属医院科研项目(No.2022008)
北京中医药大学“解码中医”揭榜挂帅项目(No.2023-JYB-JBZD-037)。
