葛根素抑制人成骨样MG-63细胞凋亡的分子机制研究  

A Study on Molecular Mechanism of Puerarin Inhibiting Apoptosis of Human Osteoblastic MG-63 Cells

作　　者：董瓅瑾[1] 高丹丹 杨静[2] 郭小芹[2] 王越[2] 金鑫[4] DONG Li-jin;GAO Dan-dan;YANG Jing;GUO Xiao-qin;WANG Yue;JIN Xin(Editorial Department of Journal,Logistics College of Chinese People's Armed Police Forces,Tianjin 300309, China;Staff Room of Pathogenic Biology,Logistics College of Chinese People's Armed Police Forces, Tianjin 300309, China;Staff Room of Military Medicine,Logistics College of Chinese People's Armed Police Forces, Tianjin 300309, China;Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China)

机构地区：[1]武警后勤学院学报编辑部,天津300309 [2]武警后勤学院病原生物学教研室,天津300309 [3]天津中医药大学,天津300192 [4]武警后勤学院军事药学教研室,天津300309

出　　处：《解放军医药杂志》2019年第4期16-21,共6页Medical & Pharmaceutical Journal of Chinese People’s Liberation Army

基　　金：国家自然科学基金项目(81273520;81572852);天津市自然科学基金项目(12JCZDJC2630;18JCZDJC332000);武警后勤学院科学技术研究项目(WHJ201712;WHJ201713)

摘　　要：目的研究葛根素抑制人成骨样MG-63细胞凋亡的分子机制。方法建立顺铂诱导的成骨细胞凋亡模型,DAPI染色法检测葛根素对MG-63细胞凋亡的影响,RT-PCR和蛋白免疫印迹法分别检测凋亡相关因子Bcl-xL mRNA和蛋白的表达水平,应用ER阻断剂ICI 182,780和小RNA干扰技术探讨葛根素调节成骨细胞凋亡的作用机制。结果顺铂10μmol/L组MG-63细胞凋亡率高于对照组,葛根素+顺铂组低于顺铂10μmol/L组(P <0. 05)。不同浓度葛根素组MG-63细胞Bcl-xL mRNA和蛋白表达水平均高于对照组(P <0. 05)。葛根素+ICI+顺铂组成骨细胞凋亡率高于葛根素+顺铂组(P <0. 05)。葛根素+顺铂组MG-63/scrambled、MG-63/ERα和MG-63/ERβshRNA细胞凋亡低于顺铂组(P <0. 05)。结论葛根素抑制顺铂诱导的人成骨样MG-63细胞凋亡作用是经ER途径,由ERα和ERβ共同介导的。Objective To study molecular mechanism of Puerarin inhibiting apoptosis of human osteoblastic MG-63 cells. Methods Apoptotic models of osteoblasts induced by Cisplatin were established. Effect of Puerarin on apoptosis of MG-63 cells was detected by DAPI staining. Expressions of Bcl-xL mRNA and protein were detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blot methods respectively. Mechanism of Puerarin regulating apoptosis of osteoblastic cells was investigated by using estrogen receptor (ER) blocking agent ICI 182, 780 and small RNA interference technology. Results Apoptotic rate of MG-63 cells in 10 μmol/L Cisplatin group was significantly higher than that in control group, and the rate in Puerarin+Cisplatin group was significantly lower than that in 10 μmol/L Cisplatin group ( P <0.05). Expressions of Bcl-xL in MG-63 cells in different concentrations Puerarin groups were significantly higher than that in control group ( P <0.05). Apoptotic rate of osteoblasts in Puerarin+ICI+Cisplatin group was significantly higher than that in Puerarin+Cisplatin group ( P <0.05). Apoptotic rates of MG-63/scrambled, MG-63/ERα and MG-63/ERβ shRNA in Puerarin+Cisplatin group were significantly lower than those in Cisplatin group ( P <0.05). Conclusion Puerarin inhibiting Cisplatin-induced apoptosis in human osteoblastic MG-63 cells is through ER pathway and mediated by both ERα and ERβ.

关 键 词：葛根素 成骨细胞 细胞凋亡 受体 雌激素 

分 类 号：R681[医药卫生—骨科学]