摘要
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，1×10^4／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1000strain的牵张应力；B组采用2000strain的牵张应力；C组采用3000strain的牵张应力；D组作为空白对照，采用0strain的牵张应力。4组牵张应力的刺激时间均为12h。牵张应力刺激12h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）ug/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2000strain的牵张应力促进成骨细胞骨钙素基因表达和增殖，3000strain则抑制成骨细胞骨钙素基因表达和增殖。
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 （MTF） 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 osteocalein 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） ug/L respectively. The ratio of osteoblast 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 osteoealein mRNA, level of osteocalein and ratio of osteoblast proliferation in group B and group C （ P 〈 0. 05 ）. There was no significant defferenee between group A and group D （ P 〉 0. 05 ）. Conclusion It demonstrates that tensile force of 2000 strain increases significantly the expression of osteoealcin mRNA and proliferation of osteoblast. Tensile force of 3000 strain declines significantly the expression of osteoealcin mRNA and the proliferation.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2012年第6期1163-1166,共4页
Chinese Journal of Experimental Surgery
基金
基金项目：国家十二五科技支撑计划资助项目（2012BA133806）
关键词
牵张应力
成骨细胞
骨钙素
基因表达
骨延长
Strain
Osteoblast
Osteocalcin
Expression of gene
Bone lengthening
