基于SIRT6/NRF2通路研究健脾固肾化瘀方治疗糖尿病肾脏病的机制  

The Mechanism of Jianpi Gushen Huayu Decoction(健脾固肾化瘀方)in the Treatment of Diabetic Kidney Disease Based on SIRT6/NRF2 Pathway

作　　者：刘珍[1] 董礼 陈晓婷 张辉 潘保朝 苏秀海[1] 王庆海[1] 吕树泉[1] LIU Zhen;DONG Li;CHEN Xiaoting;ZHANG Hui;PAN Baochao;SU Xiuhai;WANG Qinghai;LYU Shuquan(Cangzhou Hospital of Integrated Traditional Chinese Medicine and Western Medicine of Hebei Province,Cangzhou Hebei 061000,China)

机构地区：[1]河北省沧州中西医结合医院,河北沧州061000

出　　处：《中医药导报》2025年第1期32-36,41,共6页Guiding Journal of Traditional Chinese Medicine and Pharmacy

基　　金：河北省中医药管理局2023年度中医药类科学研究课题计划项目(2023257)。

摘　　要：目的:研究健脾固肾化瘀方(JPGS)治疗糖尿病肾脏病的效果并从沉默调节蛋白6(SIRT6)/核因子类红细胞相关因子2(NRF2)通路介导的氧化应激与炎症反应研究JPGS治疗糖尿病肾脏病的作用机制。方法:60只C57BL/6J小鼠适应性喂养1周后,随机分为正常组(n=50)及造模组(n=50)。正常组小鼠给予正常小鼠维持饲料,造模组小鼠高糖高脂饮食喂养8周。8周后,造模组小鼠腹腔注射STZ进行造模。模型成功的小鼠随机分为模型组、SIRT6激动剂组、JPGS低剂量组、JPGS中剂量组及JPGS高剂量组,每组10只。采用试剂盒测定小鼠24 h尿蛋白定量,血清肌酐、尿素氮。对肾脏同一部位进行苏木素-伊红、Masson染色。采用试剂盒测定小鼠肾组织丙二醛水平,超氧化物歧化酶和谷胱甘肽过氧化物酶活性、活性氧水平、白细胞介素-1β、IL-6、肿瘤坏死因子-α水平。采用Western blotting检测SIRT6/NRF2通路相关因子[SIRT6、NRF2、血红素氧合酶-1、谷氨酸半胱氨酸连接酶催化亚基、NAD(P)H醌氧化还原酶1]表达,采用RT-qPCR检测GCLC mRNA、NQO1 mRNA、SOD1 mRNA、GPX1 mRNA表达。结果:JPGS能降低糖尿病肾脏病小鼠的Cr、BUN、24 h-UPQ,减轻肾组织病理损伤,降低MDA、ROS水平,增强SOD、GSH-PX活性,降低IL-1β、IL-6、TNF-α水平,升高SIRT6、NRF2、HO-1、GCLC、NQO1表达水平,升高GCLC mRNA、NQO1 mRNA、SOD1 mRNA及GPX1 mRNA的表达。结论:JPGS可以激活SIRT6/NRF2通路抑制氧化应激与炎症反应以缓解糖尿病肾脏病的肾损伤。Objective:To evaluate the effect of Jianpi Gushen Huayu decoction(JPGS)in the treatment of diabetic kidney disease and to explore the mechanism from the oxidative stress and inflammatory mediated by sirtuin-6(SIRT6)/nuclear factor-erythroid 2 related factor 2(NRF2)pathway.Methods:After one week of adaptive feeding,60 C57BL/6J mice were randomly divided into normal group(n=50)and modeling group(n=50).Rats were given maintenance feed in normal group,while carbohydrate and high fat diet(HFD)in modeling group for 8 weeks.After 8 weeks,mice in modeling group were received intraperitoneal injection of STZ for modeling.The successfully modeled mice were randomly divided into model group,SIRT6 agonist group,low-dose JPGS group,medium-dose JPGS group,and high-dose JPGS group,10 mice in each group.Kit was used to determine 24-h urine protein quantification(24 h-UPQ),serum creatinine(Cr)and urea nitrogen(BUN)in mice.Hematoxylin-eosin and Masson staining were performed on the same part of the kidney.The malondialdehyde(MDA),superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),reactive oxygen species(ROS),IL-1β,IL-6 and TNF-αin kidney tissue were measured by kits.Western blotting was used to detect the expression of SIRT6/NRF2 pathway-related factors [SIRT6, NRF2, HO-1, GCLC, NQO1], and RT-qPCR was used to detect the expression of GCLC mRNA, NQO1 mRNA, SOD1 mRNA, and GPX1 mRNA. Results: JPGS reduced Cr, BUN, 24h-UPQ, attenuated renal histopathological injury, decreased MDA and ROS levels, enhanced SOD and GSH-Px activities, lowered IL-1β, IL-6, and TNF-α levels, elevated the expression levels of SIRT6, NRF2, HO-1, GCLC, and NQO1, and increased expression of GCLC mRNA, NQO1 mRNA, SOD1 mRNA and GPX1 mRNA in mice of diabetic kidney disease. Conclusion: JPGS can inhibit oxidative stress and inflammatory by activating SIRT6/NRF2 pathway to alleviate DKD kidney injury.

关 键 词：糖尿病肾脏病 健脾固肾化瘀方 氧化应激 炎症 SIRT6 NRF2 

分 类 号：R285.5[医药卫生—中药学]