Lactuside B对大鼠脑缺血再灌注损伤大脑皮质bcl-2、bax mRNA及其蛋白表达的影响
Effect of lactuside B on the expression of bcl-2 and bax mRNA and their protein in rats’ cerebral cortex after cerebral ischemia-reperfusion injury
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摘要:
探讨高翅果菊根茎中主要成分lactuside B对脑缺血再灌注损伤大鼠大脑皮质bcl-2、bax mRNA及其蛋白表达的影响。建立大鼠大脑中动脉缺血再灌注损伤模型, 各组给予相应的实验药物, 分别在缺血再灌注24和72 h处死所需动物, 采用TTC染色法检测脑梗死体积, 逆转录多聚酶链反应和免疫组化二步法分别检测bcl-2、bax mRNA及其蛋白在大鼠大脑皮质的表达情况。结果表明, lactuside B各剂量组均可缩小大鼠脑皮质的梗死体积, 提高bcl-2 mRNA的表达、降低bax mRNA的表达, 且12.5和25 mg·kg−1剂量组bcl-2、bax mRNA的比值较高, 与模型组比较有显著性差异 (P < 0.05或P < 0.01); 总体上12.5和25 mg·kg−1剂量组作用较好, 优于阳性对照药尼莫地平 (P < 0.05或P < 0.01), 用药72 h的脑梗死体积、bcl-2、bax mRNA与24 h比较有显著性差异 (P < 0.05或P < 0.01)。另外, bcl-2和bax蛋白的表达情况与其基因表达趋势基本一致。以上结果提示, lactuside B可能通过上调bcl-2 mRNA增加其蛋白的表达和下调bax mRNA减少其蛋白的表达, 在一定程度上发挥较好的抗脑缺血作用。
Abstract:This study is to investigate the effect of the major chemical composition in rhizome of Pterocypsela elata, lactuside B, on expression of bcl-2, bax mRNA and their protein in rats’ cerebral cortex after cerebral ischemia-reperfusion injury. First, middle cerebral artery ischemia-reperfusion injury model was established, and each group was treated with the corresponding medicines. Animals were separately sacrificed at 24 h and 72 h. The brain infarct volumes were detected by TTC dye, bcl-2 and bax mRNA expression was checked by RT-PCR, and the proteins of bcl-2 and bax were explored by two-step immunohistochemistry in cerebral cortex of rats. Lactuside B can reduce brain infarct volume of cerebral cortex of rats, increase the expression of bcl-2 mRNA and decrease that of bax mRNA. Moreover, the ratio of bcl-2 to bax mRNA is higher in 12.5 and 25 mg·kg−1 dose group, respectively, which is significantly different from that of model group (P < 0.05 or P < 0.01). Generally, either 12.5 or 25 mg·kg−1 dose group is better than positive control medicine nimodipine (P < 0.05 or P < 0.01). In addition, the expression of bcl-2 and bax protein is consistent with their gene expression. Infarct volume and the ratio of bcl-2 to bax mRNA expression are significantly different (P < 0.05 or P < 0.01) between 72 h and 24 h group. The results demonstrated that lactuside B could play a good role in resisting cerebral ischemia by upregulating the expression of bcl-2 mRNA and protein and downregulating that of bax mRNA and protein.
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