Objective To investigate the role of optic atrophy (0PA1) in human renal tubular cells (HK-2) injured by postasphyxial-serum in neonates and effects of recombinant human erythropoietin on it. Methods The experiment was designed as control group, asphyxia group and rhEPO-treated group. The serum of neonates at 24 hours after asphyxia (Apgar Scoring less than 4) was applied as attacking factor. The morphological change of HK-2 cells was observed under inverted microscope, the cell viability was measured by CCK-8 methods, the expression of 0PA1 in cytoplast was detected by using of immunohistochemical method, the morphological change of mitochondria was observed under transmission electron microscope (TEM). Results Under inverted microscopy, HK-2 cells in control group grew very quickly. The morphological change of HK-2 was more serious in asphyxia group than that in control group. The morphological change of HK-2 was improved after the treatment of rhEPO. The cell viability (OD) was decreased in the asphyxia group (0.41 ± 0.06, P < 0.05). After the treatment of rhEPO, the cell viability (OD) was obviously increased (0.60 ± 0.08, P < 0.05). The expression of 0PA1 (OD) was significantly increased in asphyxia group (0.21 ± 0.02, P < 0.05). After the treatment of rhEPO, the expression of 0PA1 was obviously decreased (0.36 ± 0.02, P < 0.05). Under TEM, mitochondria in asphyxia group appeared extensive swelling and vacuolar degeneration with less matrix and obscure or vanished mitochondria cristae; but in control and rhEPO group, mitochondrial structure was integrated, with uniform matrix and visible mitochondria cristae. Conclusions RhEPO could relieve the injury of renal tubular cells induced by postasphyxial-serum in neonate. The pretreatment with rhEPO could increase the expression of 0PA1, then improve the morphological change of mitochondria induced by postasphyxial-serum.%目的 探讨视神经萎缩症蛋白(OPA1)在新生儿窒息后血清诱导人近曲肾小管上皮细胞(HK-2)凋亡中的表达变化及重组人促红细胞生成素(rhEPO)干预的影响.方法 将HK-2分为对照组、窒息组和rhEPO组,以新生儿重度窒息后24 h血清作为攻击因素,光镜及投射电子显微镜下观察HK-2线粒体形态变化,CCK-8法检测HK-2细胞活力,免疫组化检测HK-2的OPA1表达.结果 对照组及rhEPO组HK-2生长良好,窒息组HK-2发生凋亡相关改变.与对照组HK-2的细胞活力(0.74 ± 0.08)相比,窒息组细胞活力(0.41 ± 0.06)明显降低,而rhEPO组细胞活力(0.60 ± 0.08)则较窒息组提高,差异均有统计学意义(P均＜ 0.05).与对照组HK-2的OPA1表达(0.47 ± 0.02)相比,窒息组细胞OPA1表达(0.21 ± 0.02)显著降低,而rhEPO组(0.36 ± 0.02)较窒息组有所升高,差异均有统计学意义(P均＜ 0.05).对照组和rhEPO组HK-2的线粒体结构完整,可见线粒体嵴;窒息组HK-2的线粒体嵴消失,膜肿胀,空泡变性.结论 rhEPO可能通过增加OPA1表达而抑制线粒体分裂,减轻新生儿窒息后血清诱导HK-2细胞凋亡.
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