论文部分内容阅读
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。
Objective To investigate the molecular mechanisms by which rhein-piperizinyl-estrone (named as LC) regulates the secretion of IL-6 by human osteoblast-like MG-63 cells. Methods Based on the previous work, we selected two human osteoblast-like MG-63 cells expressing both estrogen receptor (ER) subtypes as experimental models, and detected by ELISA, RT-PCR and luciferase reporter assay , Small RNA interference and Western blotting to explore the effect of LC on the expression of bone resorption-regulated factor IL-6 induced by human osteoblasts and its mechanism. Results LC could inhibit the expression of IL-6 and the transcriptional activity of IL-6 promoter in MG-63 cells. This effect could be completely blocked by ICI182,780, a pure ER blocker. LC- Inhibition of cellular IL-6 production is mediated by ERα and ERβ. PD98059 (MEK1 / 2 inhibitor) and Wortmannin (PI3K inhibitor) blocked LC-induced activation of ERK and Akt, respectively. Conclusion LC inhibits the production of IL-6 by osteoblasts through the ER pathway mediated by both ERα and ERβ. LC can also activate osteoblasts through activation of Ras / MEK / ERK and PI3K / Akt signaling pathways.