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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。
Background: The impact of weightlessness on bone metabolism in aerospace settings is considered to be one of the most damaging effects on the human body. OBJECTIVE: To investigate the effect of Qianggu Fangzongwei Prescription on osteoblasts cultured in vitro under simulated weightlessness, and to reveal the molecular biological effects of Qiangfang Kanguizhi Prescription on bone loss. Design: Randomized controlled observation. Unit: Institute of Aerospace Medicine. MATERIALS: Experiments were performed at Beijing Aerospace Medical Engineering Research Institute from April to December in 1997. Choose SD suckling mice. Drugs: Strong bone anti-wilt (Rehmannia, Drynaria, turtle shell, Achyranthes, etc.). Methods: 15 suckling mice were randomly divided into 5 groups with 3 rats in each group. Osteoblasts of suckling rat calluses were isolated and cultured in vitro. The cells were divided into five groups: normal control group, weightlessness model group, strong osteoporosis group (2.0%), medium (1.0%) and small (0.5%) dose group. Set gyrator, 30r / min, continuous gyroscope 60h simulated weightlessness observed on the side of the cyclotron 12,30,60 h rat osteoblasts alkaline phosphatase and osteocalcin generation and alkaline phosphatase gene transcription (mRNA) Impact. MAIN OUTCOME MEASURES: The effect of Qianggu Kang Wei Decoction on alkaline phosphatase activity, osteocalcin content and alkaline phosphatase mRNA expression in osteoblast culture medium in vitro under simulated weightlessness conditions. Results: All the 15 suckling mice were included in the result analysis without any loss in the experiment. ① Compared with the normal control group, alkaline phosphatase activity of osteoblasts in model group decreased significantly at 12, 36 and 60 h (P <0.05), and the difference was significant at 12 and 60 h [(45.01 ± 12.50) and (96.18 ± 12.34), (13.17 ± 5.33), (137.36 ± 137.86) nkat / L, P <0.05]. The alkaline phosphatase mRNA expression was low and the activity of osteocalcin decreased at 60h (30.3 ± 2.75 ), (42.0 ± 10.0) μg / L, P <0.05]. Compared with the model group, alkaline phosphatase activity of cell culture fluid in different dosage groups of Chinese herbal compound at different time points increased to different extents (143.70 ± 123.86, , (54.51 ± 13.17); (156.53 ± 133.69) nkat / L, P <0.05]. The alkaline phosphatase mRNA expression activity was also higher. However, osteocalcin activity did not change significantly. Conclusion: This recipe can improve the function of osteoblasts under simulated weightlessness, relieve the differentiation inhibition and promote the formation and maturation of bone matrix.