基于SIRT3/β-catenin/PPARγ信号通路探讨丹皮酚对急性心肌梗死模型大鼠的治疗作用

Therapeutic Effect of Paeonol on Acute Myocardial Infarction Model Rats Based on SIRT3/β-catenin/PPARγSignaling Pathway

目的探讨丹皮酚对急性心肌梗死(AMI)模型大鼠的治疗作用及对沉默信息调节因子3(SIRT3)/β-连环蛋白(β-catenin)/过氧化物酶体增殖物激活受体γ(PPARγ)信号通路的影响。方法将50只无特定病原体(SPF)级雄性AMI模型SD大鼠随机分为模型组、卡托普利组、丹皮酚低剂量组、丹皮酚中剂量组和丹皮酚高剂量组,每组10只;另将10只健康雄性SD大鼠作为对照组。除对照组外,其余各组均采用左冠状动脉前降支结扎法建立AMI模型。建模成功后,卡托普利组及丹皮酚各剂量组分别灌胃10 mg/kg的卡托普利和4 mg/kg、8 mg/kg、16 mg/kg的丹皮酚;对照组和模型组给予等体积生理盐水,连续2周,每日1次。末次给药后次日,超声心动图测定大鼠心脏功能;苏木精-伊红(HE)染色观察心肌组织病理学变化;全自动生化分析仪检测血清肌酸激酶(CK)、肌酸激酶同工酶(CK-MB)、乳酸脱氢酶(LDH)、心肌肌钙蛋白T(cTnT)和心肌肌钙蛋白I(cTnI)水平;酶联免疫吸附(ELISA)法测定血清白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)含量及心肌组织丙二醛(MDA)、超氧化物歧化酶(SOD)及谷胱甘肽过氧化物酶(GSP-Px)水平;免疫印迹法(Western Blot)检测心肌组织SIRT3、β-catenin、PPARγ表达水平。结果丹皮酚可改善心肌细胞水肿,抑制心肌病理性改变。与模型组比较,丹皮酚各剂量组左室射血分数(LVEF)、左室短轴缩短率(LVFS)增加(P<0.05或P<0.01),左室质量指数(LVMI)降低(P<0.05),血清CK、CK-MB、LDH及cTnT、cTnI水平均减低(P<0.05或P<0.01),血清IL-1β、IL-6及TNF-α水平均减低(P<0.05或P<0.01),心肌组织SOD、GSP-Px水平均增加(P<0.01)、MDA降低(P<0.05),心肌SIRT3和β-catenin相对表达量均增加(P<0.05),PPARγ相对表达量均降低(P<0.05)。结论丹皮酚可改善AMI大鼠心功能,抑制氧化应激及炎症反应,其作用可能与调控心肌组织SIRT3/β-catenin/PPARγ信号通路有关。

Objective To explore the effect of paeonol on acute myocardial infarction(AMI)rats and silent information regulator 3(SIRT3)/β-catenin(β-catenin)/peroxisome proliferator-activated receptorγ(PPARγ)signaling pathway.Methods Fifty specific pathogen-free(SPF)male SD rats were randomly divided into model group,captopril group,paeonol low-dose group,paeonol medium-dose group,and paeonol high-dose group.10 rats in each group;10 healthy male SD rats were set as the control group except for the control group,the AMI models were established by ligation of left anterior descending coronary artery.After successful modeling,the captopril and paeonol groups were gavaged 10 mg/kg captopril and 4 mg/kg,8 mg/kg and 16 mg/kg paeonol respectively,and the control group and the model group were gavaged an equal volume of normal saline for 2 weeks,once a day.The next day after the last administration,the cardiac function of rats was detected by echocardiography.The pathological changes of myocardial tissue were observed by hematoxylin-eosin(HE)staining.Serum creatine kinase(CK)and creatine kinase were detected by automatic biochemical analyzer.The levels of CK isoenzyme(CK-MB),lactate dehydrogenase(LDH),cardiac troponin T(cTnT),and cardiac troponin I(cTnI)were detected.The levels of interleukin-1β(IL-1β),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α)content and myocardial tissue malondialdehyde(MDA),superoxide dismutase(SOD)and glutathione peroxidase(GSP-Px)were detected by enzyme-linked immunosorbent assay(ELISA).The levels of SIRT3,β-catenin and PPARγin the myocardial tissue were detected by Western Blot.Results Paeonol could improve myocardial edema and inhibit myocardial pathological changes.Compared with AMI model rats,left ventricular ejection fraction(LVEF),and left ventricular short-axis shortening rate(LVFS)were increased(P<0.05 or P<0.01),left ventricular mass index(LVMI)was decreased(P<0.05),and serum CK,CK-MB,LDH and cTnT,cTnI levels in each dose group of paeonol were decreased(P<0.05 or P<0.01),serum IL-1β,IL-6,and TNF-αlevels were decreased(P<0.05 or P<0.01),and myocardial tissue SOD and GSP-Px levels were increased(P<0.01),MDA was decreased(P<0.05),the expressions of SIRT3 andβ-catenin in myocardium were increased(P<0.05),and the expressions of PPARγwere decreased(P<0.05).Conclusion Paeonol could improve cardiac function in AMI rats,inhibit oxidative stress and inflammatory response,and its effects might be related to regulating the activity of SIRT3/β-catenin/PPARγsignaling pathway in myocardial tissue.