骨碎补总黄酮对晚期糖基化终末产物作用下成骨细胞分化及p38和ERKl/2蛋白激酶表达的影响

目的 研究骨碎补总黄酮(TFDF)对晚期糖基化终末产物(AGEs)作用下成骨细胞分化活性及对细胞外信号调节蛋白激酶(ERK1/2)和p38丝裂原活化蛋白激酶(p38MAPK)信号蛋白的影响,探讨骨碎补总黄酮防治骨质疏松的作用机理.方法 二次酶消化法分离培养新生SD大鼠颅骨成骨细胞以及活性观察；pNPP、ELISA、茜素红染色法分别检测不同质量浓度晚期糖基化终末产物对成骨细胞碱性磷酸酶活性(ALP)、Ⅰ型胶原(Col I)、骨钙素(BGP)以及矿化能力的影响,并观察骨碎补总黄酮对晚期糖基化终末产物作用下成骨细胞分化的干预；Westem-hlot检测骨碎补总黄酮对晚期糖基化终末产物作用下,成骨细胞p38和ERK1/2蛋白磷酸化的影响.结果 晚期糖基化终末产物(200、400、800 mg/L)可以降低成骨细胞表达ALP、Col I、BGP,降低其矿化能力.骨碎补总黄酮可以剂量依赖性的提高晚期糖基化终末产物(400 mg/L)作用下成骨细胞表达ALP、Col I、BGP和矿化能力.骨碎补总黄酮(50 mg/L)可以提高晚期糖基化终末产物(400 mg/L)作用下p38和ERK1/2的蛋白磷酸化.结论 骨碎补总黄酮可以提高晚期糖基化终末产物作用下成骨细胞分化和矿化能力,其机制可能和提高p38和ERK1/2信号蛋白的磷酸化有关.

Effects of total flavonoids in Drynaria fortunei on osteoblasts differentiation and expression of p38 and ERK1/2 signaling protein in relation to advanced glycation end products in vitro

AIM To observe the effects of total flavonoids in Drynaria fortunei(TFDF) on osteoblasts differentiation in the treatment with advanced glycation end products(AGEs),and to observe the expression of p38MAPK and ERK1/2 signaling protein in osteoblasts.METHODS Primary osteoblasts of newborn Sprague-daweley rats were extracted and cultured and their biological characteristics were observed.pNPP,ELISA,Alizarin dying were used to test ALP,TypeⅠcollagen,osteocalcin and mineralization of osteoblasts in the treatment with different concentration of AGEs respectively and after treatment with different concentration of TFDF and AGEs(400 mg/L).Western-blot was used to detect p38 and ERK1/2 protein phosphorylation after treatment by TFDF and AGEs(400 mg/L).RESULTS The levels of ALP,TypeⅠcollagen,osteocalcin and mineralization of osteoblasts in the treatment with 400 mg/L AGEs were less than those in the control group in a dose-dependent manner.TFDF(50 mg/L) increased protein phosphorylation of p38 and ERK1/2 of osteoblasts associated with AGEs(400 mg/L).CONCLUSION TFDF can increase the differentiation and mineralization of osteoblasts which were decreased by AGEs,where protein phosphorylation of p38 and ERK1/2 is concerned.