摘要
目的观察木犀草素(Lut)对慢性心力衰竭(CHF)大鼠心脏功能及心肌纤维化的改善作用,并探讨AMP活化的激酶(AMPK)/沉默信息调节因子3(SIRT3)通路的作用。方法采用缩窄腹主动脉法建立大鼠CHF模型,分组为CHF组、CHF+Lut低剂量(Lut-L)组、CHF+Lut高剂量(Lut-H)组和CHF+Lut-H+AMPK抑制剂(CC)组,另设假手术(Sham)组,每组10只。分别灌胃给予生理盐水、25 mg/kg Lut、100 mg/kg Lut、100 mg/kg Lut+0.2 mg/kg CC、生理盐水,连续8周。超声心动图检查心功能;苏木素伊红(HE)和马森(Masson)染色观察心肌组织形态;ELISA法检测心肌中肿瘤坏死因子(TNF)-α、白介素(IL)-6、丙二醛(MDA)、IL-1β、乳酸脱氢酶(LDH)、单核细胞趋化蛋白1(MCP-1)及谷胱甘肽(GSH)水平;Werstern blot检测Smad2/3、Ⅰ型胶原(COL-Ⅰ)、Ⅲ型胶原(COL-Ⅲ)、p-Smad2/3、AMPK、核因子κB(NF-κB)、p-NF-κB、p-IκBα、IκBα、锰超氧化物歧化酶(MnSOD)、p-AMPK、SIRT3蛋白表达。结果与Sham组相比,CHF组、CHF+Lut-H+CC组心肌纤维排列混乱无序,可见炎症细胞等损伤,胶原蛋白沉积分数、LVESD、LVEDP、LVEDD、MDA、LDH、TNF-α、IL-1β、IL-6、MCP-1水平及p-Smad2/3/Smad2/3、COL-Ⅰ、COL-Ⅲ、p-NF-κB/NF-κB、p-IκBα/IκBα、核/质NF-κB蛋白水平增高(P<0.05),LVSP、LVFS、LVEF、LVPWd、SOD、GSH水平及p-AMPK/AMPK、SIRT3、MnSOD蛋白水平降低(P<0.05)。与CHF组相比,CHF+Lut-L组、CHF+Lut-H组上述损伤减轻,胶原蛋白沉积分数、LVESD、LVEDP、LVEDD、MDA、LDH、TNF-α、IL-1β、IL-6、MCP-1水平及p-Smad2/3/Smad2/3、COL-Ⅰ、COL-Ⅲ、p-NF-κB/NF-κB、p-IκBα/IκBα、核/质NF-κB蛋白水平降低,CHF+Lut-H组均低于CHF+Lut-L组(P<0.05);LVSP、LVFS、LVEF、LVPWd、SOD、GSH水平及p-AMPK/AMPK、SIRT3、MnSOD蛋白水平升高,CHF+Lut-H组均高于CHF+Lut-L组(P<0.05)。与CHF+Lut-H组相比,CHF+Lut-H+CC组心肌损伤加重,胶原蛋白沉积分数、LVESD、LVEDP、LVEDD、MDA、LDH、TNF-α、IL-1β、IL-6、MCP-1水平及p-Smad2/3/Smad2/3、COL-Ⅰ、COL-Ⅲ、p-NF-κB/NF-κB、p-IκBα/IκBα、核/质NF-κB蛋白水平升高(P<0.05),LVSP、LVFS、LVEF、LVPWd、SOD、GSH水平及p-AMPK/AMPK、SIRT3、MnSOD蛋白水平降低(P<0.05)。结论Lut可能通过活化AMPK/SIRT3通路,上调MnSOD抗氧化蛋白表达,抑制NF-κB入核活化,减轻心肌氧化应激和炎症,降低CHF大鼠心肌纤维化,改善其心功能。
This study was designed to observe the therapeutic effect of luteolin(Lut)on heart function and myocardial fibrosis in rats with chronic heart failure(CHF),and explore the roles of AMP-activated kinase(AMPK)/silent information regulator 3(SIRT3)pathway in the treatment.A rat CHF model was established by the method of constricting the abdominal aorta.The model rats were divided into CHF group,CHF+Lut low-dose(Lut-L)group,CHF+Lut high-dose(Lut-H)group and CHF+Lut-H+CC(AMPK suppressant)group,with 10 rats in each group,and another sham operation(Sham)group was prepared as control.Echocardiography was used to check heart function;hematoxylin-eosin(HE)and Masson stainings were used to observe myocardial tissue morphology;ELISA method was used to detect the levels of myocardial tumor necrosis factor(TNF)-α,interleukin(IL)-6,malondialdehyde(MDA),IL-1β,lactate dehydrogenase(LDH),monocyte chemoattractant protein 1(MCP-1)and glutathione(GSH);Western blot was used to detect the expression of Smad2/3,typeⅠcollagen(COL-Ⅰ),typeⅢcollagen(COL-Ⅲ),p-Smad2/3,AMPK,nuclear factorκB(NF-κB),manganese superoxide dismutase(MnSOD),p-AMPK,SIRT3 proteins.Compared with the Sham group,the arrangement of myocardial fibers in the CHF group and the CHF+Lut-H+CC group was chaotic and disordered,with inflammatory cells and other injuries,and the collagen deposition fraction,the levels of LVESD,LVEDP,LVEDD,MDA,LDH,TNF-α,IL-1β,IL-6,MCP-1 and the protein levels of p-Smad2/3/Smad2/3,COL-Ⅰ,COL-Ⅲ,p-NF-κB/NF-κB,p-IκBα/IκBαand nuclear/plasma NF-κB were increased(P<0.05),while LVSP,LVFS,LVEF,LVPWd,SOD,GSH levels and p-AMPK/AMPK,SIRT3,MnSOD protein levels decreased(P<0.05).Compared with CHF group,the above-mentioned injury reduced in CHF+Lut-L group and CHF+Lut-H group,and the indexes mentioned above reversed,especially in CHF+Lut-H group(P<0.05).In CHF+Lut-H+CC group,AMPK suppressant CC could block these effects of Lut(P<0.05).Taken togetherm Lut may activate AMPK/SIRT3 pathway,up-regulate the expression of MnSOD antioxidant protein,inhibit the activation of NF-κB into the nucleus,reduce myocardial oxidative stress,inflammation and reverse myocardial fibrosis in CHF rats,and improve their heart function.
作者
吕芳
李卫萍
田朝霞
李锦平
LYU Fang;LI Weiping;TIAN Chaoxia;LI Jinping(Fenyang College of Shanxi Medical University,Lyuliang 032200,China)
出处
《免疫学杂志》
CAS
CSCD
北大核心
2022年第5期407-415,共9页
Immunological Journal
基金
吕梁市重点研发项目(2020SHFZ35)。
