摘要
目的探讨利多卡因调节高迁移率族蛋白B1(HMGB1)/Toll样受体4(TLR4)信号通路对慢性心力衰竭大鼠的心脏保护作用。方法将大鼠分为正常组、模型组、利多卡因低剂量组、利多卡因高剂量组和EP(HMGB1抑制剂)组,采用异丙肾上腺素皮下注射制备慢性心力衰竭大鼠模型。超声检测各组大鼠心功能;ELISA试剂盒测定血清心钠肽(ANP)、脑钠肽(BNP)、白细胞介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)水平变化;HE染色和Masson染色观察大鼠心脏组织病理学变化;RT-qPCR检测HMGB1 mRNA和TLR4 mRNA表达;Western blot检测HMGB1、TLR4、NF-κB、p-NF-κB、Bcl-2、Bax相关蛋白表达。结果正常组大鼠HE染色结果显示心肌细胞形态结构正常,Masson染色结果显示心肌组织致密有序排列,组织间没有胶原蛋白沉积。与正常组相比,模型组大鼠心肌细胞褶缩,形态结构排列不规则,有较多炎性细胞浸润,组织间有大量胶原蛋白沉积,心肌纤维化程度较重,大鼠左室舒张末期内径(LVEDD)和左室收缩末期内径(LVESD)水平、心钠肽(ANP)、脑钠肽(BNP)、IL-1β和TNF-α水平、HMGB1 mRNA和TLR4 mRNA水平、HMGB1、TLR4、p-NF-κB/NF-κB、Bax蛋白表达水平显著升高,左室射血分数(LVEF)和左室短轴缩短率(LVFS)水平、Bcl-2蛋白表达水平显著降低(P<0.05);与模型组相比,利多卡因低、高剂量组和EP组大鼠心肌细胞损伤显著改善,心肌组织中胶原蛋白显著减少,心肌纤维化程度显著减轻,大鼠LVEDD、LVESD、ANP、BNP、IL-1β和TNF-α水平、HMGB1 mRNA和TLR4 mRNA水平、HMGB1、TLR4、p-NF-κB/NF-κB、Bax蛋白表达水平显著降低,LVEF和LVFS水平、Bcl-2蛋白表达水平显著升高(P<0.05);与利多卡因高剂量组相比,EP组大鼠各项指标差异无统计学意义(P>0.05)。结论利多卡因可抑制HMGB1/TLR4信号通路,降低慢性心力衰竭大鼠的炎症损伤,改善其心功能。
Objective To investigate the cardioprotective effect of lidocaine on chronic heart failure in rats by regulating the high mobility group B1(HMGB1)/Toll like receptor 4(TLR4)signaling pathway.Methods The rat model of chronic heart failure was established by subcutaneous injection of isoproterenol.The rats were grouped into normal group,model group,low-dose lidocaine group,high-dose lidocaine group and EP(HMGB1 inhibitor)group.Ultrasound was applied to detect the cardiac function of rats in each group.ELISA was used to measure the levels of serum atrial natriuretic peptide(ANP),brain natriuretic peptide(BNP),IL-1βand TNF-α.HE staining and Masson staining were applied to observe the histopathological changes of rat heart.RT-qPCR was applied to detect the expression of HMGB1 and TLR4 mRNA.Western blot was applied to detect the expression of HMGB1,TLR4,NF-κB,p-NF-κB,Bcl-2,and Bax related proteins.Results HE staining showed normal morphology and structure of myocardial cells in the normal group,while the Masson staining showed dense and orderly arrangement of myocardial tissue,with no collagen deposition between tissues.Compared with the normal group,the myocardial cells of the model group rats were convoluted,irregularly arranged in morphology and structure,with more inflammatory cell infiltration,a large amount of collagen deposition between tissues,and a heavier degree of myocardial fibrosis.Meantime,the levels of LVEDD and LVESD,the levels of ANP,BNP,IL-1βand TNF-α,the levels of HMGB1 and TLR4 mRNA,the expression of HMGB1,TLR4,p-NF-κB/NF-κB,and Bax proteins in rats were obviously increased,but the levels of LVEF and LVFS,and the expression of Bcl-2 protein were obviously reduced in the model group(P<0.05).Compared with the model group,the myocardial cell damage in the low-dose,high-dose lidocaine groups,and EP group was obviously improved,the collagen protein in myocardial tissue was clearly reduced,and the degree of myocardial fibrosis was obviously reduced.Meantime,the levels of LVEDD and LVESD,ANP,BNP,IL-1βand TNF-α,the levels of HMGB1 and TLR4 mRNA,and the expression of HMGB1,TLR4,p-NF-κB/NF-κB,and Bax proteins were obviously reduced,while the levels of LVEF and LVFS,and the expression of Bcl-2 protein were obviously increased(P<0.05).Compared with the high-dose lidocaine group,there was no statistically obvious difference in various indicators of the EP group rats(P>0.05).Conclusion Lidocaine can inhibit the HMGB1/TLR4 signaling pathway,reduce inflammatory damage,and improve cardiac function in rats with chronic heart failure.
作者
冯昌盛
钱朵
耿媛
FENG Changsheng;QIAN Duo;GENG Yuan(Affiliated Hospital of North Sichuan Medical College,Nanchong 637000,China)
出处
《广东药科大学学报》
CAS
2023年第4期25-30,共6页
Journal of Guangdong Pharmaceutical University
基金
四川省卫生和计划生育委员会科研课题(21PJ128)。
