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目的探讨细胞凋亡与肺缺血再灌注损伤的关系以及红花注射液的干预及机制。方法健康日本大耳白兔84只,随机分为对照组、缺血再灌注1、3、5h组和红花干预1、3、5h组。复制在体肺缺血再灌注损伤模型。采用电镜和原位缺口末端标记法观测肺组织细胞凋亡情况,并计算凋亡指数;免疫组织化学和原位杂交技术检测各组肺组织Bcl-2和Bax基因表达的变化。结果缺血再灌注组肺组织细胞凋亡指数和Bcl-2、Bax蛋白及mRNA均显著高于对照组(P<0.01)。红花干预组肺组织凋亡指数、Bax蛋白及Bax mRNA低于缺血再灌注组,而Bcl-2蛋白、Bcl-2mRNA以及Bcl-2与Bax的比值较缺血再灌注组上调(P<0.01或P<0.05)。电镜观察发现,缺血再灌注组肺毛细血管内皮细胞和Ⅱ型肺泡上皮细胞超微结构损伤明显,红花干预组损伤明显减轻。肺组织凋亡指数与Bax蛋白、Bax mRNA呈显著正相关(r分别=0.926,0.913;均P<0.01),与Bcl-2/Bax蛋白、Bcl-2/Bax mRNA的比值呈负相关(r分别=-0.367,-0.375;均P<0.01)。结论肺组织细胞凋亡参与了肺缺血再灌注损伤的发生,红花注射液可能通过下调Bax基因的表达,提高Bcl-2/Bax的比值抑制肺组织细胞凋亡,从而减轻肺缺血再灌注损伤。
Objective To investigate the relationship between apoptosis and lung ischemia-reperfusion injury and the intervention and mechanism of safflower injection. Methods Eighty-four healthy Japanese rabbits were randomly divided into control group, ischemia reperfusion group 1, 3, 5 h group and safflower intervention group 1, 3, 5 h group. Replicate models of lung ischemia reperfusion injury. Electron microscopy and in situ nick end labeling were used to observe the apoptosis of lung tissues, and the apoptosis index was calculated. The changes of Bcl-2 and Bax gene expression in lung tissues were detected by immunohistochemistry and in situ hybridization. Results The apoptosis index, Bcl-2, Bax protein and mRNA in the lung tissue in the ischemia-reperfusion group were significantly higher than those in the control group (P<0.01). The apoptotic index, Bax protein and Bax mRNA in lung tissue of safflower intervention group were lower than those in ischemia-reperfusion group, while the ratios of Bcl-2 protein, Bcl-2 mRNA and Bcl-2 to Bax were higher than those in ischemia-reperfusion group (P< 0.01 or P<0.05). Electron microscopy revealed that the ultrastructure of pulmonary capillary endothelial cells and type II alveolar epithelial cells in the ischemia-reperfusion group was significantly damaged, and the damage in the safflower intervention group was significantly reduced. The lung tissue apoptosis index was positively correlated with Bax protein and Bax mRNA (r=0.926, 0.913; all P<0.01), and negatively correlated with the ratio of Bcl-2/Bax protein and Bcl-2/Bax mRNA (r Respectively =-0.367, -0.375; all P < 0.01). Conclusions Apoptosis of lung tissue is involved in the occurrence of lung ischemia-reperfusion injury. Safflower injection may reduce the expression of Bax gene and increase the ratio of Bcl-2/Bax to inhibit the apoptosis of lung tissue, thus reducing the lung ischemia. Perfusion injury.