CAJ | 학술논문

目的探讨甲状旁腺激素(parathyroid hormone,PTH)对大鼠成骨细胞增殖及相关细胞因子的调节作用.方法将传代的成骨细胞分为对照组、PTH持续给药组、PTH间歇给药组.持续给药组48 h中持续给药;PTH间歇给药组前6 h给予PTH,后42 h撤除;对照组予溶媒液.每48 h为一循环,持续作用8个循环,每个循环结束时,MTT法测定细胞增殖,检测碱性磷酸酶(ALP)活性,实时荧光定量PCR法(RT-PCR)测定胰岛素样生长因子-1(IGF-1)、骨形态发生蛋白-2(BMP-2)的表达水平.结果间歇给药组细胞增殖在第7、8个循环时明显增加;而持续给药组在第5个循环时达到峰值,随后逐渐低于其他两组,在第7、8个循环时抑制作用明显.1～4个循环时3组间ALP活性差异无统计学意义,第5～8个循环ALP活性显著增加;持续给药组第6～8个循环对ALP活性有明显抑制作用.间歇给药组细胞IGF-1 mRNA的表达在第6个循环时达到高峰后有所下降,但始终明显高于其他两组;持续给药组前期IGF-1 mRNA表达水平较高,而后期则明显低于对照组.间歇给药组BMP-2 mRNA表达水平持续升高;持续给药组前3个循环表达水平较高,随给药时间的增长呈下降趋势.结论间歇给予一定浓度的PTH可刺激成骨细胞的增殖与分化,后期作用大于前期.给药前期的ALP水平主要与IGF-1 mRNA表达水平相关,而后期则可能是通过BMP-2 mRNA表达的升高使ALP活性增强.
목적 탐토갑상방선격소(parathyroid hormone,PTH)대대서성골세포증식급상관세포인자적조절작용.방법 장전대적성골세포분위대조조、PTH지속급약조、PTH간헐급약조.지속급약조48 h중지속급약;PTH간헐급약조전6 h급여PTH,후42 h철제;대조조여용매액.매48 h위일순배,지속작용8개순배,매개순배결속시,MTT법측정세포증식,검측감성린산매(ALP)활성,실시형광정량PCR법(RT-PCR)측정이도소양생장인자-1(IGF-1)、골형태발생단백-2(BMP-2)적표체수평.결과 간헐급약조세포증식재제7、8개순배시명현증가;이지속급약조재제5개순배시체도봉치,수후축점저우기타량조,재제7、8개순배시억제작용명현.1～4개순배시3조간ALP활성차이무통계학의의,제5～8개순배ALP활성현저증가;지속급약조제6～8개순배대ALP활성유명현억제작용.간헐급약조세포IGF-1 mRNA적표체재제6개순배시체도고봉후유소하강,단시종명현고우기타량조;지속급약조전기IGF-1 mRNA표체수평교고,이후기칙명현저우대조조.간헐급약조BMP-2 mRNA표체수평지속승고;지속급약조전3개순배표체수평교고,수급약시간적증장정하강추세.결론 간헐급여일정농도적PTH가자격성골세포적증식여분화,후기작용대우전기.급약전기적ALP수평주요여IGF-1 mRNA표체수평상관,이후기칙가능시통과BMP-2 mRNA표체적승고사ALP활성증강.
Objective The effects of parathyroid hormone(PTH)on proliferation of osteoblast and the related cytokines expression were investigated.Methods The osteoblasts were divided into three groups according to the mode of exposure to PTH:intermittent exposure group, continuous exposure group,and vehicle-treated group.In the intermittent exposure group,the cells were exposed to PTH for the first 6 h of each 48 h incubation cycle, and then cultured in the absence of PTH during the remainder of the cycle.In the continuous exposure group.the cells were exposed to PTH for 48 h.Every 48 h was considered as a cycle.At each end of cycle, the cells were harvested to measure MTT value and ALP activity.mRNA level of IGF-1 and BMP-2 were determined by RT-PCR.Results MTF results showed the cells proliferation increased in the intermittent exposure group at 7th and 8th cycles.In the continuous exposure group,the proliferation of osteoblasts reached peak at 5th cycle,and decreased greatly at 7th and 8th cycles.ALP activity of three groups had no significant difference in 1-4 cycles.ALP activity of the group increased greatly in 5-8 cycles,but that of the continuous exposure group decreased greatly.The IGF-1 mRNA expression in the intermittent exposure group reached peak at the 6th cycle.Its expression level decreased gradually after that,although its level was always the highest among the three groups.In the continuous exposure group,IGF-1 mRNA expression level was higher during the early cycles, but decreased in the later cycles.BMP-2 mRNA expression level in the intermittent exposure group increased gradually.The BMP-2 expression level in the continuous exposure group was higher during the 1-3 cycles,but decreased greatly in 6-8 cycles.Conclusion PTH intermittent treatment can enhanced osteoblast proliferation and differentiation at the later period.The increase of ALP activity is related to IGF-1 and BMP-2 in early or later phase of proliferation separately.