CAJ | 학술논문

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2 mRNA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1 h后开始表达环氧酶2 mRNA、并诱导产生前列腺素E2,作用3 h时环氧酶2 mRNA的表达达到峰值,而前列腺素E2的产生量在作用后6 h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.
배경:대우부혈소판혈장촉진조직재생적이상혈소판농도、나사성분담당중요작용이급통과하충궤제발휘작용등기출문제목전환불시흔청초.목적:관찰세조혈소판대소서성골세포주——MC3T3-E1적증식급기산생전렬선소E2작용적상관성.방법:장종건강성인남성지원자신상채집병제비적세조혈소판경반복액담동용후작용우소서성골세포주MC3T3-E1,분별가입체적분수5%-15%적세조혈소판、부혈소판혈장、핍혈소판혈장혹화기타양품(소염통、종류배사인자α화전화생장인자β억제제SB431542)배양.채용세포화전렬선소E2측정시제합측정세포증식여전렬선소E2적생성량,채용RT-PCR측정배양매2 mRNA적표체.결과여결론:체적분수5%세조혈소판작용우MC3T3-E1세포1 h후개시표체배양매2 mRNA、병유도산생전렬선소E2,작용3 h시배양매2 mRNA적표체체도봉치,이전렬선소E2적산생량재작용후6 h체도봉치(40.5μg/L).수착체적분수적승고,세조혈소판대MC3T3-E1세포증식적촉진작용축점강저,병차당기체적분수체도15%시정현대MC3T3-E1세포증식적현저억제작용,이세조혈소판대MC3T3-E1세포산생전렬선소E2적작용수착기농도적배비증가이현저증강.첨가소염통회명현억제5%세조혈소판대MC3T3-E1세포증식급전렬선소E2산생적촉진작용,이첨가종류배사인자α(100μg/L)칙회명현증대세조혈소판대MC3T3-E1세포산생전렬선소E2적촉진작용.령외, SB431542(15μmol/L)가명현억제체적분수5%적세조혈소판대MC3T3-E1세포증식급전렬선소E2산생적촉진작용.제시세조혈소판촉진MC3T3-E1증식여기유도해세포생성전렬선소E2유밀절적상관성.
BACKGROUND:Platelet rich plasma can reactivate the bone tissue, while some basic problems are stil 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 cel 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. Cel quantification kit and prostaglandin E2 assay kit were applied to detect cel 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 cel s 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 cel s was decreased with the concentration increased. Washed platelet at 15%inhibited the MC3T3-El cel proliferation. However, prostaglandin E2 production of MC3T3-E1 cel s was promoted with the washed platelet concentration increased. Indomethacin significantly inhibited the effect of 50 moL/L washed platelet on MC3T3-E1 cel 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 cel s. Moreover, SB431542 (15μmol/L) significantly inhibited the promotive effect of 5%washed platelet on MC3T3-E1 cel proliferation and prostaglandin E2 production. These results suggest that washed platelet promoting MC3T3-El proliferation is closely related to prostaglandin E2 production