大黄酸哌嗪雌酚酮抑制人成骨样MG-63细胞分泌IL-6分子机制的研究  

Study on the molecular mechanism for inhibitory effects of rhein-piperizinyl-estrone on IL-6 secretion in human osteoblast-like MG-63 cells

作　　者：朱雅琴[1,2] 孙维甲[3] 杨震[1] 祁琳[4] 毛立群[5] 王越[1,4] 

机构地区：[1]天津市职业与环境危害生物标志物重点实验室,300162 [2]武警总部烟台疗养院,264003 [3]武警总医院医务部计划科,北京10003 [4]武警后勤学院免疫学教研室,天津300162 [5]武警后勤学院微生物学教研室,天津300162

出　　处：《免疫学杂志》2012年第8期669-674,共6页Immunological Journal

基　　金：国家自然科学基金项目(81041071);中国博士后科学基金面上项目(20080441340);武警医学院科学技术研究面上;青年基金项目(WYM201105;WY200902;WYQ201105)

摘　　要：目的探讨大黄酸哌嗪雌酚酮(rhein-piperizinyl-estrone,命名为LC)调节人成骨样MG-63细胞分泌IL-6的分子机制。方法在原工作基础上,选择兼有2种雌激素受体(estrogen receptor,ER)亚型表达的人成骨样MG-63细胞为研究模型,采用ELISA、RT-PCR及荧光素酶报告基因检测、小RNA干扰及免疫印迹等技术,探讨LC对人成骨细胞产生的骨吸收调节因子IL-6表达的作用及作用机制。结果 LC可抑制MG-63细胞IL-6表达和IL-6基因启动子的转录活性,该作用可被纯ER阻断剂ICI182,780完全阻断,应用小RNA干扰技术进一步证实LC对成骨细胞IL-6产生的抑制作用是由ERα和ERβ共同介导的。PD98059(MEK1/2抑制剂)和Wortmannin(PI3K抑制剂)可分别阻断LC诱导的成骨细胞ERK和Akt的活化作用。结论 LC抑制成骨细胞产生IL-6是经ER途径、由ERα和ERβ共同介导的,LC还可通过活化Ras/MEK/ERK和PI3K/Akt信号通路对成骨细胞发挥作用。This study aimed to investigate the molecular mechanism for inhibitory effect of rhein-piperizinylestrone （LC） on IL-6 secretion in human osteoblast-like MG-63 cells. Based on our previous studies, human osteoblast-like MG-63 cells containing two estrogen receptors （ER） isoforms were selected as model for this study. Enzyme-linked immnosorbent assay （ELISA）, RT-PCR, lueiferase reporter assay, small interfering double-stranded RNAs （siRNA） technology and Western blot were performed to investigate the effect of LC on IL-6 expression in osteoblast-derived cells and underline the molecular mechanism. We found that LC inhibited IL-6 expression and IL-6 promoter activity of human osteoblastic MG-63 cells. But treatment with the ER antagonist ICI 182,780 abrogated the above actions of LC on osteoblast-derived cells. In conclusion, the effects of LC on IL-6 production are mediated by both ERα and ERβ. Furthermore, LC functioned at least partially through activation of Ras/MEK/ ERK and PI3K/Akt signaling.

关 键 词：大黄酸哌嗪雌酚酮 成骨细胞 IL-6 雌激素受体 Ras/MEK/ERK和PI3K/Akt通路 

分 类 号：R737.33[医药卫生—肿瘤] R737.9[医药卫生—临床医学]