【摘要】 目的:通过体外培养成骨细胞(osteoblast,OB),观察巴戟天多糖对细胞凋亡的保护作用,探讨巴戟天促进成骨细胞活性的机制。方法:取巴戟天多糖和巴戟天水提液制备含药血清,用含药血清进行细胞培养。取24h内新生SD大鼠头盖骨成骨细胞,取2代培养的成骨细胞,分为对照组(培养过程中仅加入大鼠血清)、诱导凋亡组(对照组中加入全反式维甲酸)、巴戟天水提物组(诱导凋亡组中加入巴戟天水提物药物血清)、巴戟天多糖组(诱导凋亡组中加入巴戟天多糖药物血清),通过荧光显微镜观察,流式细胞术检测细胞凋亡,RT-PCR检测Bcl-2/Bax基因表达,对巴戟天多糖在细胞凋亡过程中的作用进行评价。结果:诱导凋亡组12h诱导的OB出现凋亡,细胞核或细胞质内可见致密的黄绿染色,染色质形成明亮的凝聚块,24h单个凋亡细胞与周围的细胞分离,细胞皱缩呈圆形或卵圆形,细胞变小,胞浆致密,细胞器相互靠近,染色质浓缩,形成不同形状和大小的块状。巴戟天水提物组、巴戟天多糖组凋亡率显著低于诱导凋亡组(P<0.01),且巴戟天多糖组低于巴戟天水提物组(P<0.05)。凋亡细胞Bcl-2 mRNA表达水平:对照组>巴戟天多糖组>巴戟天水提物组>诱导凋亡组。Bax mRNA表达水平:诱导凋亡组>巴戟天水提物组>对照组>巴戟天多糖组(P<0.01)。Bcl-2/Bax:对照组>巴戟天多糖组>巴戟天水提物组>诱导凋亡组(P<0.01)。结论:巴戟天在一定程度上可抑制全反式维甲酸诱导的成骨细胞凋亡,且巴戟天多糖的这种作用显著优于巴戟天水提物,说明巴戟天多糖是巴戟天抑制成骨细胞凋亡的主要成分之一。更多还原

【Abstract】 Objective:To explore the protection on apoptosis and the mechanism of promoting the cytoactive of osteoblast by Morinda Root Polysaccharide through the observations of the cultured osteoblast in vitro.Methods:Prepared blood serum with Morinda Root Polysaccharide and Morinda Root aqueous extract and cultured Osteoblast in vitro with it.The second generation osteoblasts in vitro were separated from the cranium of 24-hours newborn SD rat,which were divided into control group(adding only rat serum during cultivation),induction apoptosis group(adding trans-retinoic acid in control group),Morinda Root aqueous extract group(adding serum prepared by Morinda Root aqueous extract in induction apoptosis group)and Morinda Root Polysaccharide group(adding serum prepared by Morinda Root Polysaccharide in induction apoptosis group).Adopting fluorescence microscope,apoptosis detected by flow cytometry and gene expression of Bcl-2 and Bax detected by RT-PCR,to evaluate the effect of Morinda Root Polysaccharide on the course of osteoblast apoptosis.Results:The apoptotic rate of Morinda Root aqueous extract group and Morinda Root Polysaccharide group were significantly lower than that of induction apoptosis group(P<0.01).The apoptosis ratio of Morinda Root Polysaccharide group was lower than that of Morinda Root aqueous extract group(P<0.05).Expression level of Bcl-2 mRNA of apoptosis cell:control group> Morinda Root Polysaccharide group> Morinda Root aqueous extract group> induction apoptosis group(P<0.01).Expression level of Bax mRNA:induction apoptosis group> Morinda Root aqueous extract group> control group> Morinda Root Polysaccharide group(P<0.01).Bcl-2/Bax:control group> Morinda Root Polysaccharide group> Morinda Root aqueous extract group> induction apoptosis group(P<0.01).Conclusion:Morinda Root can inhibit the apoptosis of osteoblast induced by trans-retinoic acid in some extent.The above role of Morinda Root Polysaccharide is significant better than that of Morinda Root aqueous extract.It is indicated that Morinda Root Polysaccharide is one of the essential component of inhibiting osteoblast apotosis.更多还原