痰瘀同治调控TLR4/NF-κB/IκB通路对糖尿病大鼠心肌炎症反应的影响  

Co-Treatment of Phlegm and Stasis Improves Myocardial Inflammation in Rats with Diabetes Mellitus via TLR4/NF-κB/IκB Pathway

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

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：《中国实验方剂学杂志》2021年第22期59-64,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81774189)。

摘　　要：目的:观察痰瘀同治对糖尿病大鼠心肌Toll样受体4(TLR4)/核转录因子-κB(NF-κB)/核转录因子-κB抑制蛋白(IκB)信号通路的干预作用,探讨其改善糖尿病大鼠心肌炎症病变的相关机制。方法:选取清洁级雄性SD大鼠45只,随机分为正常组、化痰组、化瘀组、痰瘀同治组、阿拉氯胺组、模型组。除正常组外,其余各大鼠单次腹腔注射链脲佐菌素(STZ)55 mg·kg^(-1)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给予小陷胸汤(4.05 g·kg^(-1)),血府逐瘀汤(7.02 g·kg^(-1)),抵当陷胸汤(8.10 g·kg^(-1))灌胃,阿拉氯胺组每日予阿拉氯胺(3 mg·kg^(-1))灌胃,连续给药8周后麻醉取材。采用酶联免疫吸附测定法(ELISA)检测心肌组织TLR4蛋白及肿瘤坏死因子-α(TNF-α)表达水平;免疫组化法检测NF-κB p65和NF-κB抑制蛋白α(IκBα)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TLR4,NF-κB p65,IκBα及TNF-α的mRNA表达水平。结果:与正常组比较,模型组TLR4,NF-κB p65,IκBα及TNF-α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TLR4,NF-κB p65,IκBα及TNF-α蛋白和mRNA表达水平均有不同程度下降(P<0.01);组间比较发现,痰瘀同治组TLR4,NF-κB p65,IκBα及TNF-α蛋白和mRNA表达水平比化瘀组和化痰组下降更明显(P<0.05,P<0.01)。结论:痰瘀同治法能够改善糖尿病大鼠心肌炎症病变,且效果优于单独使用化痰法或化瘀法,其机制可能与抑制TLR4/NF-κB/IκB信号通路的激活有关。Objective:To observe the intervention of phlegm-stasis co-treatment on the myocardial Tolllike receptor 4(TLR4)/nuclear factor-κB(NF-κB)/nuclear factor-κB inhibitor(IκB)signaling pathway,and to investigate its mechanism in improving myocardial inflammation in rats with diabetes mellitus(DM). Method:Forty-five male SD rats of SPF grade were randomly divided into a normal group,a phlegm-resolving(Xiao Xianxiongtang,4.05 g·kg^(-1))group,a stasis-resolving(Xuefu Zhuyutang,7.02 g·kg^(-1))group,a co-treatment(Didang Xianxiong decoction,8.10 g·kg^(-1))group,an alagebrium chloride(3 mg·kg^(-1))group,and a model group. Except for normal group, the other rats was induced by a single intraperitoneal injection of55 mg·kg^(-1) streptozotocin(STZ)to establish DM model. After adaptive feeding for three weeks,the rats were treated correspondingly by gavage daily for eight weeks. Rats were sampled under anesthesia. Enzyme-linked immunosorbent assay(ELISA)was used to detect the protein expression of TLR4 and tumor necrosis factoralpha(TNF-α) in myocardial tissues. The expression levels of NF-κB p65 and IκBα were detected by immunohistochemistry. NF-κB p65,IκBα,TNF-α,and TLR4 mRNA expression levels were detected by realtime fluorescence-based quantitative polymerase chain reaction(Real-time PCR). Result: The protein and mRNA levels of TLR4,NF-κB p65,IκBα,and TNF-α were higher in the model group than those in the normal group(P<0.01). TLR4,NF-κB p65,IκBα,and TNF-α protein and mRNA expression levels were reduced to varying degrees in the groups with drug intervention as compared with those in the model group(P<0.01). The inter-group comparison revealed that the co-treatment group showed more manifest reduction in protein and mRNA expression levels of TLR4,NF-κB p65,IκBα,and TNF-α than the phlegm-resolving group and the stasis-resolving group(P<0.05,P<0.01). Conclusion: The co-treatment of phlegm and stasis can improve myocardial inflammation in DM rats,with superior effect to either the phlegm-resolving m

关 键 词：糖尿病心肌病变 痰瘀同治 抵当陷胸汤 Toll样受体4(TLR4)/核转录因子-κB(NF-κB)/NF-κB抑制蛋白(IκB)信号通路 炎症反应 

分 类 号：R2-0[医药卫生—中医学] R289