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目的探讨不同构型视黄酸[全反式视黄酸(all-trans retinoic acid,ATRA);9顺式视黄酸(9-cis-retinoicacid,9-Cis-RA)]对缺氧缺糖损伤的PC12细胞凋亡的影响,且通过何种受体途径作用。方法实验分组为:未刺激组(0μmol/L RA);1、5、10μmol/L,20μmol/L ATRA;1、5、10、20μmol/L9-CisRA。倒置显微镜观察缺氧缺糖损伤前后细胞形态变化,流式细胞术研究经不同浓度ATRA和9-Cis-RA刺激的缺氧缺糖损伤PC12细胞凋亡水平。ELISA监测ATRA和9-Cis-RA处理4 h后PC12细胞的LDH释放水平以了解其细胞膜损伤水平的差异。结果 ATRA组细胞凋亡率在1μmol/L浓度时与未刺激组无显著差异(P>0.05),从5μmol/L起随着浓度的增高凋亡率显著增高(P<0.05),在10、20μmol/L浓度组均显著高于相同浓度的9-Cis-RA组(P<0.05)和未刺激组(P<0.05);而9-Cis-RA在20μmol/L组凋亡率低于未刺激组,但无显著性差异(P>0.05)。乳酸脱氢酶(LDH)释放水平在ATRA刺激组从5μmol/L起随浓度的增高而显著性增高(P<0.01),在5、10、20μmol/L浓度均显著高于未刺激组和9-Cis-RA组(P<0.05),而在各浓度9-Cis-RA组与未刺激组均无显著性差异(P>0.05)。在RA信号系统中ATRA特异激活retinoic acid receptors(RARs),而9-Cis-RA同时激活RARs和retinoid X receptors(RXRs)。结论在缺氧缺糖损伤的PC12细胞ATRA可通过激活RARs受体途径发挥促凋亡效应,但这种效应可以被激活RXRs受体途径而抑制。
Objective To investigate the effect of different configurations of retinoic acid [all-trans retinoic acid (ATRA); 9-cis-retinoic acid (9-Cis-RA) Effects of Apoptosis on Injured PC12 Cells and via Which Receptor Pathway. Methods The experimental groups were as follows: unstimulated group (0μmol / L RA), 1,5,10μmol / L, 20μmol / L ATRA, 1,5,10,20μmol / L 9-CisRA. The morphological changes of cells before and after hypoxia-glucose deprivation were observed by inverted microscope. The apoptosis of PC12 cells induced by hypoxia-glucose deprivation (ATRA) and 9-Cis-RA was studied by flow cytometry. The level of LDH release in PC12 cells after 4-hour treatment with ATRA and 9-Cis-RA was monitored by ELISA to find out the difference in the level of cell membrane damage. Results The apoptotic rate of ATRA group was not significantly different from that of unstimulated group (P> 0.05), and the apoptosis rate of ATRA group increased significantly from 5μmol / L (P <0.05) The apoptosis rate of 9-Cis-RA in 20μmol / L group was significantly lower than that of 9-Cis-RA group (P <0.05) and non-stimulated group Stimulation group, but no significant difference (P> 0.05). The level of lactate dehydrogenase (LDH) release increased significantly from 5μmol / L in ATRA stimulation group (P <0.01), and was significantly higher in 5,10,20μmol / L ATRA stimulation group than in unstimulated group and 9 (P <0.05). However, there was no significant difference between the 9-Cis-RA group and the unstimulated group (P> 0.05). ATRA activates retinoic acid receptors (RARs) specifically in the RA signaling system, while 9-Cis-RA activates both RARs and retinoid X receptors (RXRs). Conclusion ATRA can induce apoptosis in PC12 cells exposed to oxygen and glucose deprivation, but this effect can be inhibited by activating the RXRs receptor pathway.