【摘要】 背景:航天环境中,失重对骨质代谢的影响被认为是对人体最严重的危害之一。目的:探讨复方强骨抗萎方对模拟失重条件下体外培养的成骨细胞的影响,拟揭示该方对抗骨丢失的分子生物学作用。设计:随机对照观察。单位:航天医学工程研究所。材料:实验于1997-04/12在北京航天医学工程研究所完成。选择SD乳鼠。药物:强骨抗萎方(熟地、骨碎补、龟板、怀牛膝等)。方法:乳鼠15只,随机分为5组,每组3只。分离乳鼠颅骨成骨细胞,体外培养后,分为正常对照、失重模型、强骨抗萎方大(2.0%)、中(1.0%)、小(0.5%)剂量5组。置回旋器,30r/min,连续回旋60h模拟失重,观察该方对回旋12,30,60h时大鼠成骨细胞碱性磷酸酶和骨钙素生成及碱性磷酸酶基因转录水平(mRNA)的影响。主要观察指标:强骨抗萎方对体外模拟失重条件下成骨细胞培养液中碱性磷酸酶活性、骨钙素含量及碱性磷酸酶mRNA表达的影响。结果:纳入的15只乳鼠均进入结果分析,实验中无脱失。①与正常对照组比较,模型组成骨细胞在回旋12,36,60h不同时间点的细胞培养液中的碱性磷酸酶活性均明显降低,其中12,60h时差异显著[(45.01±12.50),(96.18±12.34);(13.17±5.33),(137.36±137.86)nkat/L,P<0.05],碱性磷酸酶mRNA表达低微;骨钙素活性均减低,60h时差异显著[(30.3±2.75),(42.0±10.0)μg/L,P<0.05]。②与模型组比较,中药复方不同剂量组于回旋不同时间点细胞培养液中的碱性磷酸酶活性均有不同程度升高[其中60h的小、中、大剂量组分别为(143.70±123.86),(54.51±13.17);(156.53±133.69)nkat/L,P<0.05];碱性磷酸酶mRNA表达活性也较高;但骨钙素活性无明显变化。结论:该方能改善模拟失重条件下成骨细胞的功能,缓解其分化抑制状态,促进骨基质的形成和成熟。更多还原

【Abstract】 BACKGROUND In space environment the effect of weightlessness on bone metabolism is viewed as one of the most severe injuries to human body. OBJECTIVE To probe into the effect of compound Qianggu kangwei formula formula on osteoblasts cultured in vitro in simulated weightlessness so as to discover the actions of the formula on anti-bone loss in molecular biology. DESIGN Randomized controlled observation was designed. SETTING Institute of Space Medico-Engineering MATERIALS The experiment was performed in Beijing Institute of Space Medico-Engineering from April to December 1997 in which SD neonatal rats were employed. The medicine Qianggu kangwei formula shudi Radix Rehmanniae Praeparata gusuibu Rhizoma Drynariae guiban Plastrum Testudinis huainiuxi Radix Achyranthis Bidentatae etc. METHODS Fifteen neonatal rats were randomized into 5 groups 3 rats in each one. Skull osteoblasts of neonatal rats were separated and after cultured in vitro the osteoblasts were divided into 5 groups named as normal control weightlessness model group formula of large dosage 2.0% group of middle dosage 1.0% group and of small dosage 0.5% group. They were place in the gyroscope and rotated at 30 r/min continuously for 60 hours to simulate weightlessness and it was to observe the effects of such formula on the formations of osteoblastic alkaliphosphatase ALP and osteocalcin as well as ALP mRNA in 12 36 and 60 hours of rotation successively. MAIN OUTCOME MEASURES Effects of the formula on ALP activity osteocalcin content and ALP mRNA in osteoblasts cultured in vitro in simulated weightlessness. RESULTS Fifteen employed neonatal rats entered result analysis for all no one dropped out. ① Compared with normal control group ALP activity of osteoblasts in cell culture liquid was decreased remarkably in model group at various time spots of 12 hours 36 hours and 60 hours of rotation in which the significant difference presented in 12 hours and 60 hours 45.01±12.50 96.18±12.34 13.17±5.33 137.36±137.86 nkat/L P < 0.05 and ALP mRNA expression was weak osteocalcin activity was reduced in every time spot which presented significant difference in 60 hours 30.3±2.75 42.0±10.0 μg/L P < 0.05. ② Compared with model group in formula groups of different dosages ALP activities in cell culture liquid at various time spots were increased at different extents in which at the time spot of 60 hours the activities of small middle and large dosages were 143.70±123.86 54.51±13.17 and 156.53±133.69 nkat/L successively,P < 0.05 . ALP mRNA expression was relatively high but there was no obvious change in osteocalcin activity. CONCLUSION The formula improves osteoblastic function in simulated weightlessness releases its differentiating inhibition and promotes the formation and maturation of bone matrix.更多还原