摘要
目的探讨健骨颗粒含药血清对成骨细胞G_1期调节蛋白的影响。方法采用酶消化法培养SD大鼠成骨细胞,健骨颗粒含药血清干预,以生理盐水大鼠血清为对照,运用流式细胞术检测成骨细胞增殖周期,免疫细胞化学法、RT-PCR法检测成骨细胞G_1期调节蛋白细胞周期蛋白(Cyclin)D1、CDK4、P21表达。结果 20%健骨颗粒含药血清能推进成骨细胞增殖周期进程,促进其增殖;成骨细胞核中存在Cyclin D1、CDK4、P21等蛋白表达;与生理盐水血清比较,健骨颗粒含药血清干预24、48、72 h均能提高成骨细胞CyclinD1、CDK4蛋白及mRNA表达(P<0.05或P<0.01),而降低P21表达(P<0.05,P<0.01)。结论健骨颗粒含药血清能从mRNA及蛋白层面上提高体外培养成骨细胞Cyclin D1、CDK4表达,抑制负性调节因子p21的表达,调节G_1期调节蛋白,促进成骨细胞增殖。
Objective To investigate the effects of Jiangu granule containing serum（JGG-serum） on the cyclins in rat＇s osteoblast at G_1 phase.Methods Osteoblasts isolated by enzymatic digestion from SD rats were cultured and intervened with JGG-serum or normal saline（as control） respectively.Cell generation cycle was detected by flow cytometry,and expressions of Cyclin D1,cyclin-dependent kinase 4（CDK4）,oncogene protein （P21） in the osteoblast were detected dynamically using immuno-cytochemical and RT-PCR technique. Results As compared with the control,the cell generation cycle and cell proliferation were proceeding quicker in the JGG-serum（20%） intervention group;with higher protein and mRNA expressions of Cyclin D1 and CDK4, as well as much lowered expressions of P21 in nuclei of osteoblast detected at all time points（24 h,48 h and 72 h after treatment,P0.05 or P0.01）.Conclusion JGG-serum can adjust the G_1 phase cyclins in osteoblast cultured in vitro,increase the mRNA and protein expressions of Cyclin D1 and CDK4,and inhibit P21 expression, so as to accelerate the proliferation of osteoblast.
出处
《中国中西医结合杂志》
CAS
CSCD
北大核心
2010年第9期966-969,共4页
Chinese Journal of Integrated Traditional and Western Medicine
基金
国家自然科学基金资助项目(No.30572402)
福建省科技厅社会发展处重点资助项目(No.2009Y0028)
福建省科技厅青年人才资助项目(No.2008F3054)
陈可冀中西医结合发展基金.福建省中西医结合老年性疾病重点实验室资助项目(No.ckj2008038o2008J1004-01)
