摘要
目的探讨p38MAPK及Caspase3在高浓度左炔诺孕酮（LNG）诱导体外培养人子宫肌瘤细胞（UtLMC）凋亡过程中的作用。方法原代培养UtLMC传代后，加入不同浓度LNG，流式细胞术测定其对细胞凋亡率的影响，并以AO／EB双染法区分早、晚期凋亡细胞和坏死细胞；Western blot测定p38MAPK、Caspase3在LNG诱导细胞凋亡中的活性变化。结果流式细胞术分析，加药组细胞凋亡率随LNG浓度的增加而逐渐升高，与对照组相比有显著性差异（t值分别为8．458、25．582、58．415、127．390，均P〈0．05）；AO／EB双染后发现，对照组无明显凋亡细胞，10μg／mL LNG作用UtLMC后，早期凋亡细胞多于阴性对照组，随着LNG剂量的增加，晚期凋亡细胞也逐渐增多，核浓聚、偏位，被染成桔红色；在10μg／mL以上LNG诱导UtLMC凋亡中，p38MAPK磷酸化水平升高（t值分别为96．278、165．878，均P〈0．05），Caspase3被明显激活（t值分别为31．527、53．190，均P〈0．05）。结论高浓度LNG诱导UtLMC凋亡的机制涉及多途径、多靶点，可能与直接激活p38MAPK信号通路，诱导Caspase3活化有关。
Objective To explore the function of p38 MAPK and Caspase 3 in levonorgestrel （LNG） induced apoptosis in cultured human uterine leiomyoma cells （UtLMC） in vitro. Methods After passaging on primary cultured UtLMC, different concentrations of LNG were given. Apoptosis ratios of cultured celia were analyzed by flow cytometry and AO/EB double staining was applied to discriminate the apoptotic cells from dead ones. The western blot was used to determine phosphorylation level of p38 MAPK and Caspase 3 in LNG-induced UtLMC apoptosis. Results Flow cytometry analysis indicated that the apoptosis rate in LNG treatment group was enhanced with the increasing of LNG concentration, and there were significant differences between LNG group and control group （ t value was 8. 458, 25. 582, 58. 415 and 127. 390, respectively, all P 〈 0.05 ）. The results of AO/EB double staining showed that there was no obvious apoptosis in control group, and there were more viable apoptotic cells in 10μg/mL LNG treatment group than negative control group. With dose increasing, the number of non-viable apoptotic cells increased, and increased uptake and dislocation of nuclear in reddish yellow occurred. The phosphorylation level of p38 MAPK was up-regulated by 10μ g/mL and 20μg/mL LNG treatment （t value was 96. 278 and 165. 878, respectively, both P 〈 0.05）, and Caspase 3 was activated （t value was 31. 527 and 53. 190, respectively, both P 〈 0.05）. Conclusion The underlying molecular mechanism by which LNG participates in the induction of human UtLMC apoptosis involves multiple channels and targets. It may be due to the direct activation of p38MAPK signal pathway and induced activation of Caspase 3.
出处
《中国妇幼健康研究》
2012年第4期492-494,共3页
Chinese Journal of Woman and Child Health Research
