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目的:探讨葛花总黄酮对大鼠心肌缺血再灌注损伤的保护作用及其作用机制研究。方法:取Wistar大鼠72只,随机分为假手术组,模型组,阳性药组(复方丹参片,240 mg·kg~(-1)),葛花总黄酮高、中、低剂量组(20,40,60 mg·kg~(-1)),预防给药28 d,通过夹闭冠状动脉30 min后恢复供血再灌注的方法复制大鼠心肌缺血再灌注损伤动物模型。造模完成后,通过2,3,5-氯化三苯基四氮唑(TTC)染色测定各组大鼠心肌梗死面积,通过苏木素-伊红(HE)染色观察心肌组织病理性形态学改变,检测血清中总抗氧化能力(T-AOC),水平腺嘌呤核苷三磷酸(ATP),二磷酸腺苷(ADP),天门冬氨酸氨基转移酶(AST),乳酸脱氢酶(LDH),磷酸肌酸激酶(CPK)水平;酶联免疫吸附测定(ELISA)检测心肌细胞组织匀浆液细胞肿瘤因子-α(TNF-α),白细胞介素-6(IL-6),白细胞介素-1β(IL-1β)含量水平;ELISA检测心肌细胞组织细胞凋亡基因B细胞淋巴瘤/白血病-2(Bcl-2),Bcl-2相关X蛋白(Bax),半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)含量表达。结果:与模型组比较,葛花总黄酮高、中剂量组治疗组大鼠心肌梗死面积显著降低(P<0.05,P<0.01);葛花总黄酮高、中剂量治疗组大鼠心肌组织病变和心肌细胞凋亡状况均好转,其中葛花总黄酮高剂量组治疗组效果最为显著;葛花总黄酮高、中剂量治疗组大鼠血清中AST,CPK,ATP,ADP,LDH水平明显降低,心肌组织中TNF-α,IL-6,IL-1β含量明显降低(P<0.05,P<0.01);葛花总黄酮高、中剂量组上调大鼠心肌组织中Bcl-2含量,降低Bax,Caspase-3含量。结论:葛花总黄酮能够降低心肌梗死面积、改善组织病理学改变,提示葛花总黄酮对心肌缺血再灌注大鼠具有保护作用,其作用机制可能与葛花总黄酮能降低炎症细胞因子浸润,降低心肌组织细胞凋亡有关。
Objective: To investigate the protective effect of Pueraria lobata flavonoids on myocardial ischemia-reperfusion injury in rats and its mechanism. Methods: Seventy two Wistar rats were randomly divided into four groups: sham-operation group, model group, positive control group (240 mg · kg -1) 20, 40 and 60 mg · kg ~ (-1)) for 28 days. The rats were subjected to myocardial ischemia-reperfusion injury by means of clamping the coronary artery for 30 min and then reperfusion. After modeling, the area of myocardial infarction in each group was determined by TTC staining. Pathological changes of myocardial tissue were observed by hematoxylin-eosin staining (T-AOC), adenosine triphosphate (ATP), adenosine diphosphate (ADP), aspartate aminotransferase (AST), lactate dehydrogenase ) And creatine phosphokinase (CPK). The levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin The levels of Bcl-2, Bcl-2, Cysteinyl aspartate Caspase-3 expression. Results: Compared with the model group, the area of myocardial infarction in the Pueraria lobata flavonoids treated group was significantly decreased (P <0.05, P <0.01); the Pueraria lobata flavonoids high and medium dose treatment group myocardial tissue changes And the apoptosis status of cardiomyocytes were all improved. The effect of the high-dose Pueraria lobata flavonoids treatment group was the most significant. The levels of AST, CPK, ATP, ADP and LDH in the serum of the rats treated with Pueraria lobata flavonoids were significantly decreased, The contents of TNF-α, IL-6 and IL-1βwere significantly decreased in myocardial tissue (P <0.05, P <0.01); Pueraria total flavonoids high and medium dose groups upregulated the content of Bcl- Caspase-3 content. Conclusion: Pueraria total flavonoids can reduce myocardial infarct size and improve histopathological changes, suggesting that Pueraria total flavonoids have a protective effect on myocardial ischemia-reperfusion rats, its mechanism may be associated with Pueraria total flavonoids can reduce the infiltration of inflammatory cytokines , Reduce myocardial cell apoptosis related.