参七糖络丸激活Nrf2/HO-1/NQO1信号通路改善2型糖尿病小鼠骨骼肌氧化应激损伤  

Activation of Nrf2/HO-1/NQO1 Signaling Pathway by Shenqi Tangluo Pill Improves Oxidative Stress Injury of Skeletal Muscle of Type 2 Diabetes Mellitus Mice

作　　者：裴晓丽 梁永林[1] 段永强[2] 朱向东 宋冰 白敏[2] 赵芸慧 赵思晨 刘中唐 苏蓓蓓 PEI Xiaoli;LIANG Yonglin;DUAN Yongqiang;ZHU Xiangdong;SONG Bing;BAI Min;ZHAO Yunhui;ZHAO Sichen;LIU Zhongtang;SU Beibei(Gansu University of Chinese Medicine,Lanzhou 730000,China;Key Laboratory of Ministry of Education of Traditional Chinese Medicine Prevention and Treatment of High-Incidence Diseases in Ningxia,Yinchuan 750000,China;Gansu Industry Technology Center for Laboratory Animal,Lanzhou 730000,China)

机构地区：[1]甘肃中医药大学,兰州730000 [2]宁夏区域高发病中医药防治教育部重点实验室,银川750000 [3]甘肃省实验动物行业技术中心,兰州730000

出　　处：《中国实验方剂学杂志》2024年第7期131-139,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：宁夏回族自治区重点研发计划项目(2022BEG02034);宁夏回族自治区重点研发计划项目引才专项(2022BSB03084)。

摘　　要：目的:基于核因子E_(2)相关因子2(Nrf2)/血红素氧合酶-1(HO-1)/醌氧化还原酶1(NQO1)通路探讨参七糖络丸对2型糖尿病小鼠骨骼肌氧化应激损伤的干预作用及机制。方法:选取SPF级7周龄雄性db/db小鼠60只适应性饲养1周,按随机数字表法分为模型组、参七糖络丸低、中、高剂量组(19、38、76 g·kg^(-1)))和二甲双胍组(0.26 g·kg^(-1))灌胃,每组12只,另选12只同周龄雄性db/m小鼠为空白组。连续干预8周。检测小鼠空腹血糖(FBG);采用酶联免疫吸附测定法(ELISA)检测空腹血清胰岛素(FINS)水平,计算稳态模型胰岛素抵抗指数(HOMA-IR)、胰岛素敏感指数(HOMA-ISI);口服葡萄糖耐量实验(OGTT)、胰岛素耐量实验(ITT);生化试剂盒检测骨骼肌组织超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活性及丙二醛(MDA)、还原型烟酰胺腺嘌呤二核苷酸磷酸(NADPH)含量;苏木素-伊红(HE)染色观察各组小鼠骨骼肌组织病理学改变;免疫荧光法(IF)检测骨骼肌组织中活性氧(ROS)及4-羟基壬烯醛(4-HNE)水平;蛋白免疫印迹法(Western blot)检测骨骼肌组织中Nrf2、HO-1、NQO1、谷氨酸-半胱氨酸连接酶催化亚基(GCLC)蛋白的表达水平。结果:与空白组比较,模型组小鼠FBG、FINS、HOMA-IR均明显升高(P<0.05),HOMA-ISI降低(P<0.05);OGTT、ITT结果显示,小鼠血糖在各时间节点均明显升高(P<0.05),糖耐量和胰岛素耐量明显受损;骨骼肌组织中SOD、GSH-Px活力明显降低(P<0.05),MDA、NADPH含量明显升高(P<0.05);骨骼肌组织肌纤维排列疏松,细胞核紊乱,见炎细胞浸润;骨骼肌组织中ROS、4-HNE表达水平明显升高(P<0.05);骨骼肌组织中Nrf2、HO-1、NQO1、GCLC蛋白表达水平明显降低(P<0.05)。与模型组比较,二甲双胍组FBG、FINS、HOMA-IR均明显降低(P<0.05),HOMA-ISI升高(P<0.05);OGTT、ITT结果显示,二甲双胍组小鼠血糖在各时间节点均明显降低(P<0.05);骨骼肌组织中SOD、GSH-Px活性明显升高(P<0.05),MDObjective:To investigate the effect and mechanism of Shenqi Tangluo pill (SQTLP) on oxidative stress injury of skeletal muscle of type 2 diabetes mellitus (T2DM) mice based on nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1)/NAD(P)H quinone oxidoreductase 1 (NQO1) pathway.Method:A total of 60 7-week-old male db/db mice[specific pathogen-free (SPF) grade]were selected and fed for one week for adaption.They were divided into the model control group,SQTLP low-,medium-and high-dose (19,38,and 76 g·kg^(-1)) groups and metformin group (0.26 g·kg^(-1)) by gavage.Each group consisted of 12 mice.Twelve male db/m mice of the same age were selected as the blank group.The intervention was implemented continuously for 8 weeks.Fasting blood glucose (FBG) was detected.Fasting serum insulin (FINS) levels were detected by enzyme-linked immunosorbent assay (ELISA),and the homeostasis model assessment-insulin resistance (HOMA-IR) index and the homeostasis model assessmentinsulin sensitivity index (HOMA-ISI) were calculated.Oral glucose tolerance test (OGTT) and insulin tolerance test (ITT) were conducted.The activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) and the contents of malondialdehyde (MDA) and reduced nicotinamide adenine dinucleotide phosphate(NADPH) in skeletal muscle tissues were detected by biochemical kits.Hematoxylin-eosin (HE) staining was used to observe the pathological changes in skeletal muscle tissues.The levels of reactive oxygen species (ROS) and 4-hydroxynonenal (4-HNE) in skeletal muscle tissue were detected by immunofluorescence (IF).The expression levels of Nrf2,HO-1,NQO1 and glutamate-cysteine ligase catalytic subunit (GCLC) proteins in skeletal muscle tissues were detected by Western blot.Result:Compared with those in the blank group,FBG,FINS and HOMA-IR in the model group were significantly increased (P<0.05),while HOMA-ISI was decreased (P<0.05).The results of OGTT and ITT showed that blood glucose was significantly increased at all time points (P

关 键 词：参七糖络丸 2型糖尿病胰岛素抵抗 骨骼肌 氧化应激 核因子E_(2)相关因子2(Nrf2)/血红素氧合酶-1(HO-1)/醌氧化还原酶1(NQO1)通路 

分 类 号：R2-0[医药卫生—中医学] R33R289R587.1