刘洋　刘修恒　王磊

[摘要] 目的 探討芹黄素联合缺血后处理对大鼠肾缺血再灌注损伤的影响。 方法 将50只大鼠随机分为5组：假手术组（sham组）、缺血再灌注组（I/R组）、芹黄素组（api组）、缺血后处理组（IPO组）、芹黄素+缺血后处理组（api+IPO组）。建立大鼠肾缺血再灌注损伤模型，处死动物取肾组织石蜡包埋切片，观察肾组织病理学变化，免疫组化检测Fas、Caspase-3和Bcl-2表达，Real-time PCR检测Fas、Caspase-3、PARP-1和Bcl-2表达水平，Western blot检测Caspase-3、Bax和Bcl-2表达水平。 结果 与sham组相比，I/R组病理组织显示肾小管损伤严重；免疫组化示Fas、Caspase-3蛋白表达增强，Bcl-2表达降低（P < 0.01）；Real-time PCR示Fas、Caspase-3和PARP-1 mRNA表达明显上调，Bcl-2 mRNA表达明显下调（P < 0.01）；Western blot示Caspase-3和Bax表达水平明显升高，Bcl-2表达水平明显降低，差异有高度统计学意义（P < 0.01）。与I/R组相比，api组、IPO组和api+IPO组病理组织显示肾小管损伤显著改善；免疫组化示Fas、Caspase-3蛋白表达下调，Bcl-2表达上调（P < 0.05）；Real-time PCR示Fas、Caspase-3和PARP-1 mRNA表达明显降低，Bcl-2 mRNA表达明显增高（P < 0.05）；Western blot示Caspase-3和Bax表达水平明显下降，Bcl-2表达水平明显升高，差异有统计学意义（P < 0.05）。 结论 芹黄素和缺血后处理均能减轻肾脏缺血再灌注损伤，其机制可能与其上调Bcl-2基因和下调Bax、Caspase-3等基因表达有关，从而抑制肾缺血再灌注损伤后所造成的细胞凋亡。

[关键词] 缺血再灌注；芹黄素；缺血后处理；凋亡

[中图分类号] R364.1 [文献标识码] A [文章编号] 1673-7210（2017）05（c）-0022-04

[Abstract] Objective To investigate the effects of apigenin combined with ischemic postconditioning on renal ischemia/reperfusion injury in rats. Methods Fifty rats were randomly divided into 5 groups： sham-operated group （sham group）， ischemia/reperfusion group （I/R group）， apigenin group （api group）， ischemic postconditioning group （IPO group） and api+IPO group. The rat model with renal ischemia/reperfusion injury was established， then the animals were sacrificed and nephridial tissue was taken to make paraffin-embedded sections. The pathologic changes of nephridial tissue were observed， the expression of Fas， Caspase-3， Bax were detected by immunohistochemistry， the expression of Fas， Caspase-3， PARP-1 and Bcl-2 were detected by real-time PCR， the expression of Caspase-3， Bax and Bcl-2 were detected by Western blot. Results Compared with the sham group， in I/R group， the pathological tissue showed that kidney tubules were damaged seriously； the immunohistochemistry showed that the expression of Fas and Caspase-3 were enhanced， the expression of Bcl-2 was decreased （P < 0.01）； Real-time PCR showed that the expression of Fas， Caspase-3 and PARP-1 mRNA were up-regulated significantly， the expression of Bcl-2 was down-regulated significantly （P < 0.01）； Western blot showed that the expression of Caspase-3 and Bax were increased significantly， the expression of Bcl-2 was reduced significantly， the differences were highly statistically significant （P < 0.01）. Compared with I/R group， in api+I/R group， IPO group and api+IPO group， the pathological tissue showed that kidney tubules were alleviated； the immunohistochemistry showed that the expression of Fas and Caspase-3 were reduced， the expression of Bcl-2 was increased （P < 0.05）； Real-time PCR showed that the expression of Fas， Caspase-3 and PARP-1 mRNA were down-regulated significantly， the expression of Bcl-2 was up-regulated significantly （P < 0.05）； Western blot showed that the expression of Caspase-3 and Bax were reduced significantly， the expression of Bcl-2 was increased significantly， the differences were statistically significant （P < 0.05）. Conclusion Both apigenin and ischemic postconditioning can reduce the renal ischemia/reperfusion injury， the mechanism may be related to up-regulating the expression of Bcl-2 and down-regulating the expression of Bax， Caspase-3， so as to inhibit the cell apoptosis caused by renal ischemia/reperfusion injury.

[Key words] Ischemia/reperfusion； Apigenin； Ischemic postconditioning； Apoptosis

肾缺血再灌注损伤是指肾组织缺血时和重获血流灌注或氧供应后，器官和组织发生损伤甚至功能衰竭[1]。肾缺血再灌注损伤常常发生在严重创伤、急性失血、中毒性休克和肾移植术后，极易引发急性肾衰竭[2]。缺血后处理是指组织或器官在长期缺血后再重获多次血流灌注或氧供应的过程，过程中缺血组织和器官的内源性保护机制被启动，缓解组织和器官的缺血再灌注损伤[3]。缺血后处理能抑制肾缺血再灌注损伤诱发的肾组织细胞凋亡，但机制尚不十分清楚[4-5]。芹黄素是一种天然多酚类化合物，它以黄色素的形式广泛存在于多种植物中，能产生明显的抗氧化作用，可清除体内过多的自由基，稳定细胞膜结构[6-7]。本研究旨在探讨芹黄素联合缺血后处理方案对肾缺血再灌注损伤的相关保护作用机制。

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