雷公藤甲素对中国仓鼠卵巢细胞凋亡的影响及黄体酮的干预效果
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摘要
目的探讨雷公藤甲素对中国仓鼠卵巢(CHO)细胞凋亡的影响及黄体酮的干预效果。方法将CHO细胞分为对照组、雷公藤甲素组、黄体酮组、雷公藤甲素+黄体酮组,分别给予等量培养液、0. 05μg/μl雷公藤甲素、0. 05μg/μl黄体酮、0. 05μg/μl雷公藤甲素联合0. 05μg/μl黄体酮干预。分别于干预后6 h及12 h检测4组CHO细胞凋亡率、细胞外信号调节蛋白激酶(ERK)磷酸化及即早基因c-fos表达水平。结果雷公藤甲素组干预后6 h及12 h的细胞凋亡率均高于对照组和黄体酮组(均P <0. 05),但两个时间点的细胞凋亡率差异无统计学意义(P> 0. 05)。雷公藤甲素+黄体酮组干预后6 h的细胞凋亡率高于对照组和黄体酮组,而低于雷公藤甲素组(均P <0. 05),且干预后12 h的细胞凋亡率低于干预后6 h(P <0. 05)。雷公藤甲素组的ERK磷酸化水平低于对照组,c-fos水平高于对照组,雷公藤甲素+黄体酮组的ERK磷酸化水平和c-fos水平均低于对照组和雷公藤甲素组(均P <0. 05)。结论雷公藤甲素可促进CHO细胞凋亡,可能与其引起ERK/c-fos异常表达有关,但干预时间延长并不增加细胞凋亡程度。黄体酮对雷公藤甲素所致仓鼠卵巢细胞损害具有保护功能。
Objective To investigate the apoptosis effects of triptolide on ovary( CHO) cells and the intervention effect of progesterone in Chinese hamster. Methods CHO cells were divided into control group,triptolide group,progesterone group,or triptolide + progesterone group,and treated with equivalent nutrient solution,0. 05 μg/μl triptolide,0. 05 μg/μl progesterone,0. 05 μg/μl triptolide combine with 0. 05 μg/μl progesterone,respectively. The apoptosis rate of CHO cells,phosphorylation of extracellular signal-regulated protein kinase( ERK) and expression level of immediately early gene c-fos were detected after 6 and 12 hours of intervention. Results The apoptosis rate of CHO cells in triptolide group was higher than that in control group or progesterone group after 6 and 12 hours of intervention( all P < 0. 05),but no statistical differences in apoptosis rates of two time points were found( P > 0. 05).The apoptosis rate of CHO cells in triptolide + progesterone group was higher than that in control group and progesterone group,but lower than that in triptolide group after 6 hours of intervention( all P < 0. 05),and the rate was even lower after 12 hours of intervention( P < 0. 05). The phosphorylation level of ERK was lower,and the expression level of c-fos was higher in triptolide group than in the control,and the phosphorylation level of ERK and expression level of c-fos were lower in the triptolide + progesterone group than in the control group or triptolide group( all P < 0. 05).Conclusion Triptolide can promote the apoptosis of CHO cells,which might be related to the abnormal expression of ERK/c-fos. However,with the prolonged intervention time of triptolide,the degree of apoptosis does not increase.Progesterone has a protective function against injure of CHO cells induced by triptolide in hamster.
Objective To investigate the apoptosis effects of triptolide on ovary( CHO) cells and the intervention effect of progesterone in Chinese hamster. Methods CHO cells were divided into control group,triptolide group,progesterone group,or triptolide + progesterone group,and treated with equivalent nutrient solution,0. 05 μg/μl triptolide,0. 05 μg/μl progesterone,0. 05 μg/μl triptolide combine with 0. 05 μg/μl progesterone,respectively. The apoptosis rate of CHO cells,phosphorylation of extracellular signal-regulated protein kinase( ERK) and expression level of immediately early gene c-fos were detected after 6 and 12 hours of intervention. Results The apoptosis rate of CHO cells in triptolide group was higher than that in control group or progesterone group after 6 and 12 hours of intervention( all P < 0. 05),but no statistical differences in apoptosis rates of two time points were found( P > 0. 05).The apoptosis rate of CHO cells in triptolide + progesterone group was higher than that in control group and progesterone group,but lower than that in triptolide group after 6 hours of intervention( all P < 0. 05),and the rate was even lower after 12 hours of intervention( P < 0. 05). The phosphorylation level of ERK was lower,and the expression level of c-fos was higher in triptolide group than in the control,and the phosphorylation level of ERK and expression level of c-fos were lower in the triptolide + progesterone group than in the control group or triptolide group( all P < 0. 05).Conclusion Triptolide can promote the apoptosis of CHO cells,which might be related to the abnormal expression of ERK/c-fos. However,with the prolonged intervention time of triptolide,the degree of apoptosis does not increase.Progesterone has a protective function against injure of CHO cells induced by triptolide in hamster.
引文
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[2]崔艳秋,陆莉,王晓民.雷公藤内酯醇及其衍生物的神经保护作用及机制[J].首都医科大学学报,2011,32(6):859-864.
[3]丁樱,杨晓青,李向峰,等.中成药雷公藤多苷对儿童性腺发育的影响[J].中医儿科杂志,2013,9(1):20-23.
[4]孙凤,杨兴华,马冬梅,等.雷公藤用药者生殖毒性发生率的Meta分析[J].中国药物警戒,2014,11(2):94-99,103.
[5]崔卫东,张士巧,赵幸福,等.天癸一号联合黄体酮治疗抗精神药物所致药源性闭经的疗效及安全性研究[J].中医药导报,2015,21(17):67-70.
[6] Liu J,Jiang ZZ,Liu L,et al. Triptolide induces adverse effect on reproductive parameters of female Sprague-Dawley rats[J]. Drug Chem Toxicol,2011,34(1):1-7.
[7] Zhang J,Liu L,Mu X,et al. Effect of triptolide on estradiol release from cultured rat granulosa cells[J]. Endocr J,2012,59(6):473-481.
[8]张武,胡永良. P44/42MAPK介导雷公藤甲素致肾上腺皮质细胞凋亡作用[J].辽宁中医药大学学报,2014,16(3):27-29.
[9]王凤兰.主动免疫联合黄体酮治疗习惯性流产患者的临床疗效[J].中国药物经济学,2016,11(11):80-81.
[10]杨春桃.不同剂量黄体酮治疗无排卵型月经失调的临床效果[J].中国妇幼保健,2015,30(31):5 489-5 491.
[11]杨彩虹.黄体酮不同给药途径治疗黄体功能不全先兆流产的疗效对比分析[J].北方药学,2017,14(1):97-98.
[12]胡利霞,涂雪松,陈玉环,等.安宫黄体酮联合补肾活血方治疗抗精神药物所致的药源性闭经的疗效及安全性研究[J].实用医技杂志,2016,23(7):742-744.