健骨颗粒含药血清对大鼠成骨细胞G_1期调节蛋白的影响

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(P0.05或P0.01),而降低P21表达(P0.05,P0.01)。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。

Effects of Jiangu Granule Containing Serum on the Cyclins in Rat's Osteoblast at G1 Phase

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.