健骨颗粒通过G_1/S期调控蛋白对成骨细胞增殖的影响  

Effects of Jiangu Granules on the proliferation of osteoblasts through G_1/S phase cell-cycle regulated proteins

作　　者：贾晓康 黄云梅[1] 李超雄[2] 林煜[2] 林向全[2] 黄美雅[1] 吴银生[1] 

机构地区：[1]福建中医药大学中西医结合研究院,福建福州350122 [2]厦门大学附属福州第二医院骨科,福建福州350007

出　　处：《中国医药导报》2016年第9期9-13,共5页China Medical Herald

基　　金：国家自然科学基金资助项目(81473706)

摘　　要：目的观察健骨颗粒对成骨细胞G_1/S期调控的影响,探讨健骨颗粒促进成骨细胞增殖的作用机制。方法制备健骨颗粒血清组、模型血清组和雌二醇血清组。采用酶消化法培养SD大鼠成骨细胞,健骨颗粒含药血清干预,以模型血清和雌二醇血清为对照。运用流式细胞术检测成骨细胞增殖周期,荧光定量PCR法检测成骨细胞G_1/S期调控蛋白Cyclin E、CDK2、p21和转录因子E2F-1 m RNA的表达。结果 15%的雌二醇血清与健骨颗粒血清干预48 h,成骨细胞增殖速度均明显快于模型血清组(P<0.01);G_0/G_1期成骨细胞比例明显降低(P<0.01),S期、G_2/M期细胞比例及增殖指数则明显高于模型血清组(P<0.01);与模型血清组比较,雌二醇血清组与健骨颗粒血清组能提高成骨细胞Cyclin E、CDK2及转录因子E2F-1 m RNA的表达(P<0.01),而降低p21的表达(P<0.01)。结论健骨颗粒通过调节成骨细胞G_1/S期调控机制,推进成骨细胞顺利通过G_1/S检测点,促进成骨细胞增殖。Objective To observe the effects of Jiangu Granules on the G1/S phase cell-cycle regulated proteins of osteoblasts, so as to reveal the mechanism of Jiangu Granules in promoting osteoblasts proliferation. Methods The Jiangu Granules-serum group, model-serum group and estradiol-serum group were prepared. Osteoblasts of SD rats were cultivated by enzymatic digestion and intervened with Jiangu Granules-serum, model-serum and estradiol-serum（as control） respectively. The cell cycles were analyzed by flow cytometry, while the fluorescence quantitative PCR was applied to measure cyclin E, CDK2, p21 and E2F-1 m RNA. Results The cell proliferation rates of osteoblasts intervened with estradiol serum and Jiangu Granules-serum for 48 hours were faster than that with the same concentration model-serum（P〈0.01）. Compared with the model-serum group, the proportion of osteoblasts in the G0/G1 phase were significantly reduced after intervention with 15% of estradiol serum and Jiangu Granules-serum for 48 hours（P〈0.01）,while the proportion of cells in S phase, G2/M phase and proliferation index was much higher（P〈0.01）. The expression of cycling E, CDK 2 and transcription factor E2F-1 m RNA of osteoblasts in the Jiangu Granules-serum group and the estradiol-serum group was higher than that of model-serum group（P〈0.01）, while the expression of p21 m RNA was lower than the model-serum group（P〈0.01）. Conclusion Jiangu Granules-serum can adjust the G1/S phase cellcycle regulated mechanism in osteoblast, so as to push the cells passing G1/S checkpoint and accelerate the proliferation of osteoblast.

关 键 词：绝经后骨质疏松症 成骨细胞 细胞周期 G1/S期 健骨颗粒 

分 类 号：R285.5[医药卫生—中药学] R681[医药卫生—中医学]