【摘要】：目的觀察新型合成化合物Z-甲基-5-(3,5-二甲氧基苯甲撐基)-2-亞氨-1-咪唑-4-酮(DIMO)是否對大鼠心肌缺血再灌注損傷具有保護作用,并且探究其發(fā)揮作用的信號通路。方法在體實驗采用86只成年雄性SD大鼠建立急性大鼠在體心肌缺血再灌注(I/R)損傷模型。隨機分為4組:假手術組(Sham組)、單純缺血再灌注對照組(I/R組)、DIMO1mg/kg后處理組(DIMO-1組)、DIMO 2mg/kg后處理組(DIMO-2組)。除Sham組外,其余各組均缺血30 min,再灌注4 h。于再灌注4 h末,提取心臟,采用氯化三苯基四氮唑染色法(TTC法)測定心肌梗死范圍;采用免疫印跡法(Western blot技術)檢測Cathepsin B、Cathepsin L、Beclin-1、LC3Ⅱ/Ⅰ及P62蛋白表達水平;采用免疫共沉淀和Western blot檢測RIP1K和RIP3K的相互作用。離體實驗通過培養(yǎng)原代心肌細胞,通過氧糖剝奪(OGD)方法建立缺氧缺糖細胞模型。隨機分為4組:對照組(non-OGD組)、對照給藥組(non-OGD+DIMO組)、模型組(OGD/R組)、模型給藥組(OGD/R+DIMO組)。OGD3h,復氧12h后,采用LDH漏出率檢測心肌細胞損傷;Annexin-V和碘化丙啶(PI)雙染色法檢測心肌細胞的凋亡壞死率;免疫共沉淀和Western blot檢測RIP1K和RIP3K的相互作用;采用Western blot檢測Cathepsin B、Cathepsin L、Beclin-1、LC3Ⅱ/Ⅰ及P62蛋白表達水平。結果體內實驗:與Sham組相比,I/R組心肌梗死范圍增大,心肌細胞損傷增加,Cathepsin B、Cathepsin L、Beclin-1、LC3Ⅱ/Ⅰ及P62蛋白表達上調(P0.05),RIP1K表達上調(P0.05)。與I/R組比較,DIMO-1與DIMO-2組心肌梗死范圍減小,心肌細胞損傷減少,Cathepsin B、Cathepsin L、Beclin-1、LC3Ⅱ/Ⅰ及P62蛋白表達下調(P0.05),RIP1K表達下調(P0.05)。體外實驗:與non-OGD組比較,OGD/R組Cathepsin B、Cathepsin L、Beclin-1、LC3Ⅱ/Ⅰ及P62蛋白表達上調(P0.05),LDH漏出率和細胞凋亡壞死率增多(P0.05)。與OGD/R組比較,OGD/R+DIMO組Cathepsin B、Cathepsin L、Beclin-1、LC3Ⅱ/Ⅰ及P62蛋白表達下調(P0.05),LDH漏出率和細胞凋亡壞死率降低(P0.05)。結論新型合成化合物DIMO對大鼠在體心肌I/R損傷具有保護作用,其機制可能是通過作用于抑制壞死性凋亡及自噬水平,進而減少再灌注期間的心肌細胞死亡。
[Abstract]:Objective to investigate the protective effect of a new synthetic compound Z-methyl-5- (3O5-dimethoxybenzomethyl) -2-imidazole-4-one (DIMO) on myocardial ischemia-reperfusion injury in rats. And explore its role in the signal pathway. Methods 86 adult male SD rats were used to establish acute myocardial ischemia-reperfusion (I / R) injury model in vivo. They were randomly divided into four groups: sham operation group (Sham group), ischemia reperfusion control group (I / R group) and DIMO1mg/kg post treatment group (DIMO-1 group), DIMO 2mg/kg post treatment group (DIMO-2 group). With the exception of Sham group, all the other groups were subjected to ischemia for 30 min, and reperfusion for 4 h. At the end of 4 h after reperfusion, the heart was extracted and the myocardial infarction size was determined by triphenyl tetrazolium chloride staining (TTC), and the expression levels of Cathepsin Bon Cathepsin L, Beclin-1C 3 鈪，

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