痰瘀同治调控NOX2/p47phox/LOX通路减轻氧化应激改善糖尿病大鼠心肌纤维化  

Modulation of NOX2/p47phox/LOX Pathway by Phlegm and Stasis Reduces Oxidative Stress and Improves Myocardial Fibrosis in Diabetic Rats

作　　者：李飞翔 储全根[1,2] 喻锦 储俊[1,2] LI Fei-xiang;CHU Quan-gen;YU Jin;CHU Jun(College of Traditional Chinese Medicine,Anhui University of Chinese Medicine,Hefei,230038;Key Labratory of Xin'an Medicine,Ministry of Education,Anhui University of Chinese Medicine,Hefei,230038)

机构地区：[1]安徽中医药大学中医学院,合肥230038 [2]安徽中医药大学新安医学教育部重点实验室,合肥230038

出　　处：《中国中西医结合杂志》2024年第10期1228-1234,共7页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金资助项目(No.81774189);安徽省中医药领军人才项目(No.ZYYLJRC201911);安徽省高校优秀科研创新团队计划(No.2022AH010037)。

摘　　要：目的观察痰瘀同治对糖尿病大鼠心肌还原性烟酰胺腺嘌呤二核苷酸磷酸氧化酶2(NOX2)/p47phox/赖氨酰氧化酶(LOX)通路的干预作用,探究其通过减轻氧化应激反应,改善糖尿病大鼠心肌纤维化病变的相关机制。方法选取健康雄性SD大鼠60只,随机分为:正常组、化痰组、化瘀组、痰瘀同治组、阿拉氯胺组和模型组,每组10只。单次腹腔注射链脲佐菌素(STZ)溶液55 mg/kg建立糖尿病模型,正常喂养3周后,化痰组、化瘀组、痰瘀同治组每日分别予小陷胸汤[4.05 g/(kg·d)]、血府逐瘀汤[7.02 g/(kg·d)]、抵当陷胸汤[8.10 g/(kg·d)]灌胃,阿拉氯胺组每日予阿拉氯胺[3 mg/(kg·d)]灌胃,连续给药8周后麻醉取材。HE及Masson染色观察大鼠心肌组织病理形态变化;免疫组化法检测大鼠心肌组织锰超氧化物歧化酶(MnSOD)表达水平;实时荧光定量聚合酶链式反应(Real-time PCR)检测大鼠心肌组织NOX2、p47phox及LOX的mRNA表达水平;Western Blot法检测大鼠心肌组织NOX2、p47phox及LOX蛋白表达水平。结果与正常组比较,模型组大鼠心肌纤维排列紊乱,部分断裂,胶原纤维增生,分布不均匀;MnSOD表达显著降低(P<0.01),NOX2、p47phox及LOX蛋白和mRNA表达水平显著升高(P<0.01)。与模型组比较,各用药组大鼠心肌组织病理损害明显改善,心肌纤维排列较为整齐,胶原纤维增生程度减轻;MnSOD表达明显增强(P<0.05,P<0.01),NOX2、p47phox及LOX蛋白和m RNA表达水平均不同程度下降(P<0.05,P<0.01)。组间比较发现,痰瘀同治组较化瘀组和化痰组心肌组织病理损害改善效果更佳,MnSOD表达水平增强更明显(P<0.01),NOX2、p47phox及LOX蛋白和mRNA表达水平较化瘀组和化痰组下降更明显(P<0.05,P<0.01)。结论痰瘀同治法能够减轻糖尿病大鼠心肌组织氧化应激反应,改善心肌纤维化,且效果优于单独使用化痰或化瘀之法,其机制可能与调控NOX2/p47phox/LOX信号通路有关。Objective To observe the interventional effect of phlegm-stasis co-treatment on myocardial NOX2/p47phox/LOX pathway in diabetic rats,and to investigate the mechanism related to its reduction of oxidative stress and improvement of myocardial fibrotic lesions in diabetic rats.Methods Totally 60 male SD rats were randomly divided into normal group,resolving phlegm group,dispersing blood stasis group,resolving phlegm and dispersing blood stasis group,alagebrium chloride group and model group,10 in each group.A single intraperitoneal injection of streptozotocin 55 mg·kg^(-1)was used to establish a model of diabetes mellitus.After normal feeding for 3 weeks,in the resolving phlegm group,dispersing blood stasis group,resolving phlegm and dispersing blood stasis group,Xiaoxianxiong Decoction(4.05 g·kg^(-1)·d^(-1)),Xuefu Zhuyu Decoction(7.02 g·kg^(-1)·d^(-1))and Didang Xianxiong Decoction(8.10 g·kg^(-1)·d^(-1))were given respectively by intragastriction,and in the alagebrium chloride group,alagebrium chloride(3 mg·kg^(-1)·d^(-1))was given by intragastriction,and the material was taken under anesthesia after 8 weeks of continuous administration.Histomorphological changes in rat myocardium observed by HE and Masson staining.The expression level of manganese-containing superoxide dismutase(MnSOD)was detected by immunohistochemistry.Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)was used to detect the mRNA expression levels of NOX2,p47phox and LOX in rat myocardial tissue.The protein expression levels of NOX2,p47phox and LOX in rat myocardial tissue were measured by Western Blot.Results Compared with the normal group,myocardial fibers in the model group were disorganized,partially broken,and collagen fibers were proliferated and unevenly distributed,MnSOD expression reduced(P<0.01),NOX2,p47phox and LOX protein and mRNA expression levels increased(P<0.01).Compared with the model group,the myocardial histopathological damage of rats in each dosing group was significantly improved,the myocar

关 键 词：糖尿病心肌病变 痰瘀同治 氧化应激反应 心肌纤维化 中药复方 

分 类 号：R285.5[医药卫生—中药学]