哌芳安他对心脏缺血再灌注损伤的保护作用

Protective effects of pivanampeta on ischemia-reperfusion injury of rat heart

目的:观察哌芳安他(pivanampeta,Piv)对大鼠在体心肌缺血再灌注损伤的影响。方法:实验动物分为2组,一组为缺血30min再灌注30min组,再分为假手术组、对照组和Piv9mg·kg-1用药组,测定有关心功能指标和心肌梗死面积;另一组为缺血30min再灌注2h组,再分为假手术组、模型组以及Piv3、6和9mg·kg-1用药组,各用药组于缺血前30min静脉注射给药,再灌注2h后取心脏标本石蜡包埋后切片,免疫组化法检测Bax、Bcl2、caspase3、MMP2、和PPARγ蛋白质表达,原位杂交方法检测MMP2和PPARγmRNA表达,TUNEL法和DNA凝胶电泳观察心肌细胞凋亡。结果:(1)与对照组比较,Piv3mg·kg-1组坏死面积(nec)与缺血面积(aar)之比减少21%(P<0.01),坏死面积与左室面积(lv)之比减少22%(P<0.01)。心率、反映心脏收缩功能的指标+dpdtmax和Vmax以及反映心脏舒张功能的指标-dpdtmax分别在再灌注1min和30min明显改善(P<0.05)。(2)TUNEL法检测,Piv各亚组降低细胞凋亡作用显著(P<0.05),但DNA凝胶电泳检测,模型组、Piv3、6mg·kg-1组可见到DNA梯带,假手术组和Piv9mg·kg-1组则无。(3)免疫组化检查,Piv3、6和9mg·kg-1呈剂量依赖性减少Bax、caspase3、MMP2蛋白质和MMP2的mRNA表达,增加Bcl2、PPARγ(peroxisomeproliferatoractivatedreceptorγ)蛋白质和PPARγmRNA表达。结论:Piv预处理对心肌缺血再灌注损伤有保护作用,表现为减少心肌梗死面积、改善心功能、减少心肌细胞凋亡。

AIM: To observe the effects of pivanampeta (Piv) on ischemia-reperfusion injury of rat heart in vivo. METHEDS: Sprague-Dawley rats were randomly divided into two groups. One was 30 min reperfusion group, which was subdivided into sham, control and Piv groups with 30 min ischemia followed by 30 min reperfusion to detect some data related to cardiac function and the area of myocardium infarction. Another was 2 h reperfusion group, which was further subdivided into such sections as follows: sham, model, Piv 3, 6 and 9 mg·kg -1 groups. Apart from the sham, Piv and NS (control) groups were administered intravenously 30 min before occlusion. Then hearts were excised, paraffined and cut into 4 μm think. Immunohistochemistry, in situ hybridization, TUNEL and DNA agarose gel electrophoresis were performed to detect the expression of Bax, Bcl-2, caspase-3, MMP-2 and PPAR γ protein and MMP-2, and PPARγ mRNA. RESULTS: Compared with control group, the ratio of nec/aar after Piv injected decreased 21% (P< 0.05), while nec/lv reduced to 22%(P< 0.05).Heart rate, +dp/dt_ max and V_ max representing the systolic function of heart as well as -dp/dt_ max which was the indicator of diastole improved dramaticly at 1 min and 30 min after reperfusion, respectively(P< 0.05). In dose-dependent manner, the expression of Bax and caspase-3 protein, MMP-2 protein and mRNA depressed, while the PPARγ protein and mRNA enhanced by Piv. The apoptotic index of subgroups injected Piv reduced with important significance by TUNEL compared with model(P< 0.05). DNA ladder existed in model, Piv 3、6 mg·kg -1, rather than Piv 9 mg·kg -1 by agarose gel electrophoresis. CONCLUSION: Piv shows the protective ability against I/R injury of myocardium. Piv administration may reduce the area of MI and improve cardiac function as well as decrease apoptotic cardiomyocytes.