缺氧培养下M2型巨噬细胞上清液及杜仲总黄酮对成骨细胞生物行为学的影响  

Influence of M2 macrophage supernatant combined with Eucommia flavonoids on the biological behavior of osteoblasts under hypoxic conditions

作　　者：张文博[1] 张贤[2] 

机构地区：[1]南京中医药大学,江苏省南京市210023 [2]南京中医药大学无锡附属医院骨科,江苏省无锡市214071

出　　处：《中国组织工程研究》2017年第12期1819-1825,共7页Chinese Journal of Tissue Engineering Research

基　　金：江苏省"333"工程资助项目(BRA2012031)~~

摘　　要：背景:含成骨细胞成分的组织工程材料在骨科领域广泛应用,如何提高成骨细胞对缺氧条件的耐受能力,改善缺氧条件下成骨细胞的生物学行为,是组织工程的重要研究目标。目的:探究M2型巨噬细胞上清液和杜仲总黄酮对缺氧环境培养下的成骨细胞生物学行为的影响,为组织工程研究提供基础。方法:复苏MC3T3-E1细胞,分离、诱导及培养M2型巨噬细胞提取上清液,分别为空白对照组、M2型巨噬细胞上清液组、杜仲总黄酮组及M2型巨噬细胞上清液联合杜仲总黄酮组。空白对照组细胞不进行干预。M2型巨噬细胞上清液组、杜仲总黄酮组及M2型巨噬细胞上清液联合杜仲总黄酮组分别加入M2型巨噬细胞上清液和/或100 mg/L杜仲总黄酮。结果与结论:M2型巨噬细胞上清液组、杜仲总黄酮组及M2型巨噬细胞上清液联合杜仲总黄酮组细胞活性明显高于对照组,且M2型巨噬细胞上清液联合杜仲总黄酮组S期和G_2/M期细胞百分比显著高于对照组。M2型巨噬细胞上清液联合杜仲总黄酮组Runx2蛋白、Ⅰ型胶原蛋白、碱性磷酸酶相对mRNA水平、碱性磷酸酶相对活性以及矿化钙离子相对浓度均显著高于其他各组。提示M2型巨噬细胞上清液联合杜仲总黄酮能够促进缺氧条件下成骨细胞的增殖和分化,维持成骨细胞正常矿化功能和成骨作用,提高成骨细胞对缺氧条件的耐受能力。BACKGROUND： Tissue-engineered materials containing osteoblasts have been widely used in the treatment of orthopedic diseases. How to improve hypoxic tolerance and the biological behavior of osteoblasts under hypoxic conditions is an important goal of the research on tissue engineering.OBJECTIVE： To explore the effect of M2 macrophage supernatant and Eucommia flavonoids on the biological behaviorof osteoblasts under hypoxia to pave ways for tissue engineering. METHODS：After MC3T3E1 cells were resuscitated, and M2 macrophages were isolated, induced and cultured, all cells were divided into control, M2 macrophage, Eucommia flavonoids and combination groups. The control group received no intervention. Cells in the M2 macrophage, Eucommia flavonoids and combination groups were exposed to M2 macrophages or/and 100 mg/L Eucommia flavonoids, respectively. RESULTS AND CONCLUSION：The cell viability in the M2 macrophage, Eucommia flavonoids and combination groups was significantly higher than that in the control group. The percentage of cells in the S and G2/M phases in the combination group was significantly higher than that in the control group. The relative mRNA expression levels of Runx2, collagen type Ⅰ and alkaline phosphatase, alkaline phosphatase activity and relative concentration of calcium ions in the combination group were significantly higher than those in the other three groups. To conclude, M2 macrophage supernatant combined with Eucommia flavonoids can promote osteoblast proliferation and differentiation under hypoxic conditions, maintain normal mineralization and osteogenesis of osteoblasts, and improve the tolerance ability of osteoblasts to hypoxia.

关 键 词：组织构建 骨细胞 缺氧 M2型巨噬细胞 杜仲 总黄酮 成骨细胞 细胞增殖 成骨分化 矿化作用 

分 类 号：R318[医药卫生—生物医学工程]