CAJ | 학술논문

目的观察不同的机械牵张应力对成骨细胞骨钙素基因表达和增殖的影响,探讨骨延长的分子生物学机制.方法从新生SD大鼠颅盖骨分离培养成骨细胞,1×104/cm2密度接种至细胞加力板上,随机分为4组,每组8份.A组采用1000 strain的牵张应力；B组采用2000 strain的牵张应力；C组采用3000 strain的牵张应力；D组作为空白对照,采用0 strain的牵张应力.4组牵张应力的刺激时间均为12 h.牵张应力刺激12 h后提取成骨细胞RNA,逆转录-聚合酶链反应( RT-PCR)一步法分析成骨细胞骨钙素mRNA表达,放射免疫法测定细胞培养上清液中骨钙素含量,噻唑蓝(MTT)比色法测定成骨细胞增殖率.结果 A、B、C、D组细胞骨钙素基因表达比值分别为0.421 ±0.022、0.446±0.015、0.361±0.018、0.415±0.014；细胞培养上清液中骨钙素含量分别为(3.201 ±0.216)、(3.457 ±0.096)、(2.641 ±0.019)、(3.159±0.322) μg/L；细胞增殖率分别为(0.2869±0.0079)％、(0.2971 ±0.0105)％、(0.2778 ±0.0084)％、(0.2861±0.0125)％.B、C、D组比较,差异均有统计学意义(P＜0.05)；A组与D组比较差异无统计学意义(P＞0.05).结论 2000 strain的牵张应力促进成骨细胞骨钙素基因表达和增殖,3000 strain则抑制成骨细胞骨钙素基因表达和增殖.
목적 관찰불동적궤계견장응력대성골세포골개소기인표체화증식적영향,탐토골연장적분자생물학궤제.방법 종신생SD대서로개골분리배양성골세포,1×104/cm2밀도접충지세포가력판상,수궤분위4조,매조8빈.A조채용1000 strain적견장응력；B조채용2000 strain적견장응력；C조채용3000 strain적견장응력；D조작위공백대조,채용0 strain적견장응력.4조견장응력적자격시간균위12 h.견장응력자격12 h후제취성골세포RNA,역전록-취합매련반응( RT-PCR)일보법분석성골세포골개소mRNA표체,방사면역법측정세포배양상청액중골개소함량,새서람(MTT)비색법측정성골세포증식솔.결과 A、B、C、D조세포골개소기인표체비치분별위0.421 ±0.022、0.446±0.015、0.361±0.018、0.415±0.014；세포배양상청액중골개소함량분별위(3.201 ±0.216)、(3.457 ±0.096)、(2.641 ±0.019)、(3.159±0.322) μg/L；세포증식솔분별위(0.2869±0.0079)％、(0.2971 ±0.0105)％、(0.2778 ±0.0084)％、(0.2861±0.0125)％.B、C、D조비교,차이균유통계학의의(P＜0.05)；A조여D조비교차이무통계학의의(P＞0.05).결론 2000 strain적견장응력촉진성골세포골개소기인표체화증식,3000 strain칙억제성골세포골개소기인표체화증식.
Objective To study the effects of differential strain on the expression of osteocalcin mRNA and the proliferation of osteoblast,and to explore the molecular mechanism of bone lengthening.Methods The osteoblast was collected from newborn SD rat' s,skull and cultured,and then randomly divided into 4 groups (8 samples for each group):group A,group B,group C and group D.In group A,the cells were treated with a tensile force of 1000 strain.In group B,the cells were treated with a tensile force of 2000 strain.In group C,the cells were treated with a tensile force of 3000 strain.In group D,the cells were cultured not to add any force.The period of mechanical stimulation was 12 hours in all the four groups.The cells were collected and total RNA were obtained after being treated with strain of 12 hours.The expression of osteocalcin mRNA of osteoblast was analysed with reverse transcription-polymerase chain reaction (RT-PCR).The amount of osteocalcin in culture medium was detected by the method of radioimmunoassay.The ratio of proliferation of osteoblast was measured using methyl thiazol tetrazolium (MTT)reduction assay.Results The expression of osteocalcin mRNA in group A,B,C and D was 0.421 ±0.022,0.446 ±0.015,0.361 ± 0.018 and 0.415 ± 0.014 respectively.The level of osteocalcin in the culture medium in group A,B,C and D was (3.201 ±0.216),(3.457 ±0.096),(2.641 ±0.019) and (3.159 ±0.322) μg/L respectively.The ratio of ostenblast proliferation in group A,B,C and D was (0.2869 ±0.0079)％,(0.2971 ±0.0105)％,(0.2778 ±0.0084)％ and (0.2861 ± 0.0125)％ respectively.Compard with group D,there were significant differences of the expression of osteocalcin mRNA,level of osteocalcin and ratio of osteoblast proliferation in group B and group C ( P ＜ 0.05).There was no significant defference between group A and group D ( P ＞ 0.05 ).Conclusion It demonstrates that tensile force of 2000 strain increases significantly the expression of osteocalcin mRNA and proliferation of osteoblast.Tensile force of 3000 strain declines significantly the expression of osteocalcin mRNA and the proliferation.