人洗涤血小板促进成骨细胞增殖与前列腺素E2的作用  

Human washed platelets promote the proliferation of osteoblasts and prostaglandin E2 production

作　　者：李洪涛[1] 段建民[1] 匡威[1] 菊地宽高 片山直 

机构地区：[1]解放军广州军区广州总医院口腔科,广东省广州市510010 [2]日本明海大学齿学部,日本

出　　处：《中国组织工程研究》2012年第42期7797-7803,共7页Chinese Journal of Tissue Engineering Research

基　　金：广东省科技计划项目(2011B031800201);课题名称:改良富血小板血浆对骨髓间充质干细胞与乳牙髓干细胞体外扩增及成骨分化作用的对比实验研究~~

摘　　要：背景:对于富血小板血浆促进组织再生的理想血小板浓度、哪些成分担当重要作用以及通过何种机制发挥作用等基础问题目前还不是很清楚。目的:观察洗涤血小板对小鼠成骨细胞株——MC3T3-E1的增殖及其产生前列腺素E2作用的相关性。方法:将从健康成人男性志愿者身上采集并制备的洗涤血小板经反复液氮冻溶后作用于小鼠成骨细胞株MC3T3-E1,分别加入体积分数5%-15%的洗涤血小板、富血小板血浆、乏血小板血浆或和其他样品(消炎痛、肿瘤坏死因子α和转化生长因子β抑制剂SB431542)培养。采用细胞和前列腺素E2测定试剂盒测定细胞增殖与前列腺素E2的生成量,采用RT-PCR测定环氧酶2mRNA的表达。结果与结论:体积分数5%洗涤血小板作用于MC3T3-E1细胞1h后开始表达环氧酶2mRNA、并诱导产生前列腺素E2,作用3h时环氧酶2mRNA的表达达到峰值,而前列腺素E2的产生量在作用后6h达到峰值(40.5μg/L)。随着体积分数的升高,洗涤血小板对MC3T3-E1细胞增殖的促进作用逐渐降低,并且当其体积分数达到15%时呈现对MC3T3-E1细胞增殖的显著抑制作用,而洗涤血小板对MC3T3-E1细胞产生前列腺素E2的作用随着其浓度的倍比增加而显著增强。添加消炎痛会明显抑制5%洗涤血小板对MC3T3-E1细胞增殖及前列腺素E2产生的促进作用,而添加肿瘤坏死因子α(100μg/L)则会明显增大洗涤血小板对MC3T3-E1细胞产生前列腺素E2的促进作用。另外,SB431542(15μmol/L)可明显抑制体积分数5%的洗涤血小板对MC3T3-E1细胞增殖及前列腺素E2产生的促进作用。提示洗涤血小板促进MC3T3-E1增殖与其诱导该细胞生成前列腺素E2有密切的相关性。BACKGROUND：Platelet rich plasma can reactivate the bone tissue,while some basic problems are still unknown such as the ideal platelet concentration,the important elements and relevant mechanisms.OBJECTIVE：To investigate the correlation of washed platelet between mouse osteoblast MC3T3-E1 cell proliferation and prostaglandin E2 production.METHODS：Washed platelet was obtained from young healthy male volunteer and acted on MC3T3-E1 after repeatedly extracted from liquid nitrogen frozen.Cell quantification kit and prostaglandin E2 assay kit were applied to detect cell proliferation and PGE2 production.Reverse transcription-PCR was used to detect the cyclooxygenase-2 mRNA expression.RESULTS AND CONCLUSION：After 5% washed platelet acted on MC3T3-E1 cells for 1 hour,cyclooxygenase-2 mRNA expression and prostaglandin E2 production were promoted.Cyclooxygenase-2 mRNA expression reached a peak value at 3 hours;while prostaglandin E2 production reached a peak value at 6 hours（40.5μg/L）.The promotive effect of washed platelet on the proliferation of MC3T3-E1 cells was decreased with the concentration increased.Washed platelet at 15% inhibited the MC3T3-El cell proliferation.However,prostaglandin E2 production of MC3T3-E1 cells was promoted with the washed platelet concentration increased.Indomethacin significantly inhibited the effect of 50 moL/L washed platelet on MC3T3-E1 cell proliferation and prostaglandin E2 production.However,tumor necrosis factor-2（100μg/L） could significantly promote the effect of washed platelet on prostaglandin E2 production of MC3T3-E1 cells.Moreover,SB431542（15μmol/L） significantly inhibited the promotive effect of 5% washed platelet on MC3T3-E1 cell proliferation and prostaglandin E2 production.These results suggest that washed platelet promoting MC3T3-El proliferation is closely related to prostaglandin E2 production

关 键 词：富血小板血浆 洗涤血小板 小鼠 成骨细胞 增殖 前列腺素E2 组织工程 

分 类 号：R318[医药卫生—生物医学工程]