加减补阳还五汤通过SIRT1途径激活自噬改善糖尿病肾病小鼠肾组织损伤

Modified Buyang Huanwu Decoction improves renal tissue damage in diabetic kidney disease mice by activating autophagy through SIRT1 pathway

关键词:  糖尿病肾病  加减补阳还五汤  沉默信息调节因子1  自噬  凋亡

Key words:  diabetic kidney disease  modified Buyang Huanwu Decoction  silent information regulator 1  autophagy  apoptosis

摘要:

[目的] 探讨加减补阳还五汤是否通过沉默信息调节因子1（SIRT1）途径激活自噬改善糖尿病肾病（DKD）小鼠肾组织损伤。[方法] 将DKD小鼠随机分为模型组、中药组，db/m组小鼠作为正常组。给药8周后，检测小鼠血糖（FBG）、糖化血红蛋白（HBA1c）、总胆固醇（TC）、三酰甘油（TG）、尿素氮（BUN）、肌酐（Scr）、尿微量白蛋白（mALB）水平；实时定量反转录聚合酶连锁反应（qRT-PCR）和免疫组化检测小鼠肾组织Wilms肿瘤基因1（WT1）、nephrin及podocin的表达；蛋白质免疫印迹法（Western blot）检测小鼠肾组织SIRT1、人微管相关蛋白轻链3B（LC3B）、自噬相关蛋白7（Agt7）、自噬受体蛋白p62（P62）、Bcl-2同源结构域蛋白抗体（Beclin1）、B细胞淋巴瘤因子2（Bcl-2）、BCL-2相关X蛋白（Bax）、半胱氨酸蛋白酶-3（Caspase-3）蛋白表达；同时光镜观察肾脏病理形态学变化。[结果] 中药干预后，小鼠肾脏病理改变轻于模型组；尿微量白蛋白（mAlb）、血脂（TC、TG）下降，肾功能（BUN、Scr）改善；同时抑制了肾组织P62、Bax、Caspase-3表达，上调了SIRT1、Beclin-1、Agt7、Bcl-2、LC3B-Ⅱ/LC3B-Ⅰ的表达，并改善了肾组织足细胞标志蛋白WT1、nephrin及podocin的表达（P＜0.05或P＜0.01），但对血糖水平无明显影响（P＞0.05）。[结论] 加减补阳还五汤可通过增强DKD小鼠肾脏细胞自噬，并抑制其凋亡，保护足细胞，进而发挥肾脏保护作用。其中SIRT1途径可能是其促进自噬的潜在途径。

Abstract:

[Objective] To investigate whether the modified Buyang Huanwu Decoction can improve the renal tissue damage of diabetic kidney disease（DKD） mice by activating autophagy through SIRT1 pathway. [Methods] The mice with DKD were randomly divided into model group and traditional Chinese medicine group，and the db/m group was regarded as the normal group. After 8 weeks of administration，the levels of fasting blood glucose（FBG），glycosylated hemoglobin（HBA1c），total cholesterol（TC），triglyceride（TG），urea nitrogen（BUN），creatinine（Scr） and urinary microalbumin（mALB） were measured. The expression of wilms tumor 1 protein（WT1），nephrin and podocin in mouse kidney tissue were detected by qRT-PCR and immunohistochemistry. Western blot was used to detect the protein expressions of silent information regulator 1（SIRT）1，human microtubule-associated protein light chain 3B（LC3B），autophagy-related protein 7（Agt7），Sequestosome 1（P62），Beclin1，B-cell lymphoma-2（Bcl-2），BCL-2-associated X protein（Bax） and Caspase-3 in mice kidney tissue. At the same time，the pathological changes of kidney were observed under light microscope. [Results] After Chinese medicine intervention，the renal pathological changes in mice were lighter than those in the model group. The levels of mALB，blood lipids（TC，TG） were decreased. The renal function（BUN，Scr） was improved. At the same time，it inhibited the expressions of P62，Bax and Caspase-3 in renal tissue，up-regulated the expression of SIRT1，Beclin-1，Agt7，LC3B-Ⅱ/LC3B-I and Bcl-2. It also improved the expression of podocyte marker proteins WT1，nephrin and podocin in renal tissue（P＜0.05 or P＜0.01），but had no significant effect on blood glucose levels（P＞0.05）. [Conclusion] The modified Buyang Huanwu Decoction can exert a renal protective effect by enhancing autophagy，inhibiting apoptosis and protecting podocytes in mouse kidney tissue. Among them，the SIRT1 pathway may be its potential way to promote autophagy.