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目的研究大鼠肠缺血再灌注后肠损伤程度与聚腺苷二磷酸核糖聚合酶-1(PARP-1)激活程度的相关关系,探索肠缺血再灌注后PARP-1过度激活的水平。方法通过阻断腹腔干和肠系膜上动脉后恢复血运,制作鼠肠缺血再灌注模型,按不同缺血时间分组,再灌注2h后,处死动物取材,分别作HE染色、聚腺苷二磷酸核糖(PAR)的免疫组化染色,用TUNEL方法检测组织细胞凋亡情况,用Westernblot检测凋亡早期信号PARP-1片段p85和凋亡诱导因子(AIF)的表达情况。结果随着缺血时间的延长,肠损伤程度和PAR表达阳性率呈显著增高(P<0.05)且二者呈正相关(r=0.924,P<0.01)。PAR表达阳性率在20%左右时,凋亡早期信号PARP-1p85片段、AIF表达和TUNEL阳性率较假手术组显著升高(P<0.05);PAR表达阳性率在40%左右时,细胞坏死和组织损伤显著增加(P<0.05)。结论在大鼠肠缺血再灌注损伤中,肠损伤程度与PARP-1激活程度呈正相关,PARP-1激活率在20%左右时,组织损伤以细胞凋亡为主;PARP-1激活率在40%左右时,组织损伤以细胞坏死为主。
Objective To study the correlation between the degree of intestinal damage and the activation of PARP-1 after intestinal ischemia-reperfusion in rats and to explore the over-activation of PARP-1 after intestinal ischemia-reperfusion. Methods The model of murine intestinal ischemia-reperfusion was established by blocking the celiac trunk and the superior mesenteric artery. The rats were divided into different groups according to different ischemic time. After 2 hours of reperfusion, the animals were sacrificed and stained for HE staining. (PAR) immunohistochemical staining. TUNEL method was used to detect the apoptosis of tissue cells. Western blot was used to detect the expression of PARP-1 fragment p85 and apoptosis-inducing factor (AIF) in the early stage of apoptosis. Results With the extension of ischemic time, the degree of intestinal injury and the positive rate of PAR expression were significantly increased (P <0.05), and the two were positively correlated (r = 0.924, P <0.01). The positive rate of PARP-1p85, AIF and TUNEL in early apoptotic cells was significantly higher than that in sham operation group (P <0.05) when the positive expression rate of PAR was about 20%; while the positive rate of PAR was about 40% And tissue damage increased significantly (P <0.05). CONCLUSIONS: The degree of intestinal damage is positively correlated with the degree of PARP-1 activation in rat intestinal ischemia-reperfusion injury. When the PARP-1 activation rate is about 20%, the cell apoptosis is dominated by apoptosis. The activation rate of PARP-1 is About 40%, tissue damage mainly to cell necrosis.