益骨胶囊含药血清对晚期糖基化终末产物作用下成骨细胞分化及BMP-2和OPG的影响

Effects of Yigu Capsule Containing Serum on the Osteoblast Differentiation and the Expressions of Osteoprotegerin and Bone Morphogenetic Protein 2 after Treatment by Advanced Glycation End Products in vitro

免费下载全文查看/发表评论下载PDF阅读器

DOI：

中文关键词: 益骨胶囊成骨细胞分化骨形态发生蛋白2 骨保护蛋白

英文关键词:Yigu Capsule osteoblast differentiation bone morphogenetic protein 2 osteoprotegerin

基金项目:国家自然科学基金资助项目(No.30772885,No.81173619);广东省高等学校中药有效性与安全性重点实验室开放基金(No.kf08011);广东中医药局科研基金资助项目(No.2010118)

作者	单位
朱晓峰	暨南大学附属第一医院
王廷春	广东博济新医临床研究中心
张荣华	暨南大学药学院中药学教研室
孙升云	暨南大学附属第一医院
韩莉	暨南大学附属第一医院
王攀攀	暨南大学药学院中药学教研室
杨丽	暨南大学药学院中药学教研室

摘要点击次数: 1565

全文下载次数: 3

中文摘要:

目的研究益骨胶囊含药血清对晚期糖基化终末产物(advanced glycation end products,AGEs)作用下成骨细胞分化活性及对骨形态发生蛋白2(bone morphogenetic protein2,BMP-2)和骨保护蛋白(osteopro-tegerin,OPG)的影响,探讨益骨胶囊防治骨质疏松(osteoporosis,OP)的作用机制。方法选取10月龄SD雌性大鼠40只,随机分为益骨胶囊高、中、低剂量组及空白组,每组10只。运用大鼠灌胃的方法制备含药血清和对照血清。二次酶消化法分离培养新生SD大鼠颅骨成骨细胞,传代后分为空白对照组、模型组、中药低、中、高剂量组。模型组用AGEs(400mg/L)处理,中药低、中、高剂量组分别用AGEs(400mg/L)和低、中、高剂量含药血清共同处理,分别用pNPP、ELISA、茜素红染色法检测成骨细胞碱性磷酸酶(alkaline phosphatase,ALP)活性、Ⅰ型胶原(typeⅠcollagen,ColⅠ)、骨钙素(bone gla protein,BGP)以及矿化结节,RT-PCR测定成骨细胞中BMP-2、OPG mRNA的表达,ELISA法检测BMP-2、OPG蛋白的表达量。结果原代分离培养的新生大鼠成骨细胞具有典型的成骨细胞特征;与空白对照组比较,模型组成骨细胞(OB)ALP、ColⅠ、BGP、BMP-2、OPG蛋白及BMP-2、OPG mRNA表达降低,矿化结节减少,差异有统计学意义(P<0.01)。与模型组比较,中药各剂量组ALP、ColⅠ、BGP、BMP-2、OPG蛋白及BMP-2、OPG mRNA表达升高,矿化结节增加,且随剂量增加而明显增加,差异均有统计学意义(P<0.05,P<0.01)。结论益骨胶囊含药血清提高AGEs作用下成骨细胞分化和矿化能力,提高BMP-2、OPG的表达,这可能是益骨胶囊防治OP的机制之一。

英文摘要:

Objective To observe the effects of Yigu Capsule(YGC) containing serum on the differentiation and the expressions of osteoprotegerin(OPG) and bone morphogenetic protein 2(BMP-2) in osteoblasts after treatment by advanced glycation end products(AGEs),and to explore the mechanisms of YGC for treating osteoporosis(OP).Methods Forty 10-month-old female Sprague Dawley(SD) rats were randomly divided into four groups equally,i.e.,the low dose YGC group,the moderate dose YGC group,the high dose YGC group,and the blank control group.The YGC containing serum and the control serum were prepared with the method of gastric perfusion.Primary osteoblasts of newborn SD rats was extracted and cultured,then they were passaged and divided into five groups,i.e.,the vehicle control group,the model group,the low dose YGC group,the moderate dose YGC group,and the high dose YGC group.The model group was treated by AGEs(400 mg/L),the three YGC groups were treated by AGEs(400 mg/L) and YGC containing serum at different concentrations.Alkaline phosphatase(ALP) activity,type Ⅰ collagen(ColⅠ),bone gla protein(BGP),and mineralization of osteoblasts were tested using pNPP,ELISA,and Alizarin dyeing.The mRNA and protein levels of BMP-2 and OPG were respectively determined using RT-PCR and ELISA.Results Primary osteoblasts from newborn SD rats could be used well in this experiment.Compared with the vehicle control group,the ALP,ColⅠ,BGP,the mRNA and protein levels of BMP-2 and OPG of osteoblasts decreased in the model group,and the mineralized nodes were reduced,showing statistical difference(P<0.01).Compared with the model group,the ALP,ColⅠ,BGP,the mRNA and protein levels of BMP-2 and OPG of osteoblasts increased in the three YGC groups,and the mineralized nodes increased in a dose-dependent manner,showing statistical difference(P<0.05,P<0.01).Conclusions YGC containing serum could promote the differentiation and mineralization of osteoblasts,and improve the expressions of OPG and BMP-2 after treatment by AGEs.These might be one of YGC’s mechanisms for treating OP.

关闭