异槲皮苷对成骨细胞分化及HDAC1和Runx2表达的影响

摘要

目的:探讨异槲皮苷对成骨细胞分化及对HDAC1和Runx2表达情况的影响.方法:不同浓度梯度异槲皮苷(1×10-8,1×10-7,1×10-6,1×10-5 mol/L)连续处理MC3T3-E1细胞系,在作用第3天,借助qRT-PCR技术对MC3T3-E1细胞系中Ⅰ型胶原蛋白(type 1 collagen,Col 1)、碱性磷酸酶(alkaline phosphatase,ALP)及骨桥蛋白(osteopontin,OPN)的表达进行检测,并对骨细胞分化关键酶ALP的活性和骨钙的形成进行检测,以评估成骨细胞的分化程度.通过qRT-PCR技术检测MC3T3-E1细胞中HDAC1及Runx2的表达水平.结果:异槲皮苷能促进成骨细胞MC3T3-E1中Col 1,ALP及OPN的上调表达,且差异具有统计学意义(P＜0.05),并且呈现出一定的剂量效应.异槲皮苷诱导时,成骨细胞分化过程中ALP活性显著增强,骨钙形成明显增加.在1×10-6mol/L异槲皮苷作用下,成骨细胞系中HDAC1 mRNA和蛋白水平均显著下调(P＜0.05),而Runx2 mRNA和蛋白水平均明显上调(P＜0.05).结论:异槲皮苷可能通过下调HDAC1表达、促进Runx2表达,有效地促进成骨细胞分化,为异槲皮苷应用于治疗骨质损伤类疾病提供新的依据.%Objective:To explore effect of the isoquercetin on osteoblast differentiation and expression level of HDAC 1 and Runx2.Methods:The type 1 collagen (Co11),alkaline phosphatase (ALP) and osteopontin (OPN) expression level in MC3T3-E1 cells was detected by using qRT-PCR after treated with 1×10 8,1×10-7,1×10-6,1×10-5 mol/L isoquercetin.T-he ALP activity was assayed using ALP detection kit.Mineralization of MC3T3-E1 was evaluated by Alizarin red staining.What's more,the mRNA levels of HDAC1 and Runx2 in MC3T3-E1 cells were observed by using qRT-P CR post-treatment by 1 × 10-8,1 × 10-7,1 × 10-6,1 × 10-5 mol/L isoquercetin.Results:T-he Col 1,ALP and OPN expression level turned significantly higher after treated with 1 × 10-8,1 × 10-7,1 × 10-6,1 × 10-5 mol/L isoquercetin (P＜0.05).There was a certain dose effect between them.As the same,the ALP activity and mineralization of MC3T3-E 1 was higher post-exposure to the isoquercetin.HDAC 1 in the MC3T3-E 1 cells had lower expression level after treated with the isoquercetin (P＜0.05).However,Runx2 in the MC3T3-E1 cells had higher expression level after exposed to the isoquercetin (P＜0.05).Conclusion:The isoquercetin can promote the expression of Runx2 through inhibiting the expression of HDAC 1,in order to improve the differentiation of osteoblast.The results provide a new theoretical basis for the application ofisoquercetin in the treatment of osteoporosis.