促红细胞生成素对氯化钴诱导的PC12细胞凋亡的拮抗作用

Effect of EPO on the CoCl_2-induced apoptosis in PC12 cells

目的观察促红细胞生成素(EPO)对氯化钴(CoCl2)诱导的PC12细胞缺氧损伤凋亡的影响并探讨其作用机制。方法将PC12细胞分为四组:空白对照组、CoCl2处理组、重组人促红细胞生成素(rhE-PO)处理组、rhEPO+CoCl2处理组。应用MTT,乳酸脱氢酶(LDH),流式细胞术(FCM)及Hoechst33258染色法检测rhEPO对氯化钴诱导的细胞活性下降及凋亡的影响。通过逆转录PCR(RT-PCR)法检测Survivin表达情况。结果 MTT结果显示rhEPO预处理能够提高PC12细胞活力,0.6mmol/LCoCl2单独处理组细胞存活率仅为(43.07±5.9)%,rhEPO处理24h后细胞存活率上升明显至(77.89±3.4)%(P<0.01)。LDH检测,FCM及Hoechst33258结果表明CoCl2处理能诱导PC12细胞损伤及凋亡,而预先采用2UrhEPO处理PC12细胞可抑制CoCl2的细胞毒性作用,减少细胞缺氧性损伤及凋亡。RT-PCR显示rhEPO处理组PC12细胞Survivin表达较CoCl2处理组明显升高。结论 EPO处理能抑制CoCl2诱导的PC12细胞凋亡,其细胞保护作用的机制可能与上调PC12细胞的Survivin表达有关。

Objective To investigate the effects of erythropoietin （EPO） on cobalt chloride （ CoC12 ） -induced PC12 cells apoptosis and the possible mechanisms. Methods PC12 cells were divided into four groups： control group, CoC12 group ,rhEPO group, and rhEPO ± CoC12 group. The PC12 cells viability and toxicity were measured by MTT assay and LDH assay, respectively. Flow cytometry and Hoechst33258 staining was used to determine chromatin distribution changes of PC12 cells. The expression of Survivin mRNA was detected by reverse transcriptase polymerase chain reaction（RT-PCR）. Results MTT assay showed that the cell viability of PC12 cells treated with CoC12 was decreased to （ 43.07 ± 5.9 ） %, while rhEPO treatment increased the cell viability to （ 77.89 ±3.4 ） % （P 〈 0.01 ）. The similar results of LDH assay were observed, suggesting that rhEPO protected the PC12 cells against CoC12-induced apoptosis. Flow cytometry and Hoeehst33258 staining demonstrated that rhEF＇O treatment significantly inhibited the apoptosis of PC12 cells induced by CoC12. RT-PCR showed that rhEPO treatment induced remarkable increase in the expression of Survivin in PC12 cells cultures as compared with CoC12 treated group. Conclusions EPO protects PC12 cells from apoptosis induced by CoC12. The protective effect of EPO are executed by upregulating the expression of Survivin in PC12 cell cultures.