外周血清素对离体Sprague-Dawley大鼠成骨细胞的影响

发布时间：2018-03-17 13:12

  本文选题：血清素　切入点：5-羟色氨酸　出处：《山西医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的研究血清素与SD大鼠成骨细胞(Osteoblasts,OBs)的剂量相关性。方法(1)贴壁培养与分离新生大鼠颅骨细胞。通过观察细胞形态和荧光染色鉴定。(2)向P3代细胞培养液中混进不同浓度(10-5 mol/L、10-6 mol/L、10-7 mol/L、10-8 mol/L、10-9 mol/L、0mol/L)血清素干预。(3)第5天用MTT法分析干预因素与增殖的关系。(4)第10天分别用微量酶标法测碱性磷酸酶(Alkaline phosphatase,ALP)水平和免疫荧光法测Ⅰ型胶原蛋白(Collagen-Ⅰ,Col-Ⅰ)水平分析干预因素与分化的关系。(5)第15天用ELISA法测骨钙素(Osteocalcin,OCN)水平,分析干预因素与分化的关系。统计学方法通过SPSS 20.0收集整理并分析数据。计量资料用均数±标准差((?)±s)表示,不同组间比较采用单因素方差分析,多组间两两之间比较采用LSD-t检验,P0.05表示差异有统计学意义。结果不同浓度(10-5 mol/L、10-6 mol/L、10-7 mol/L、10-8 mol/L、10-9 mol/L、0mol/L)血清素干预,MTT法测得平均光密度值分别为:0.400±0.028,0.379±0.023,0.379±0.047,0.401±0.038,0.420±0.038,0.618±0.064,任意组与空白对照组比较,差异有统计学意义(P0.05);微量酶标法测得ALP平均水平分别为:0.046±0.002,0.017±0.002,0.026±0.002,0.046±0.002,0.051±0.002,0.073±0.003,任意组与空白对照组比较,差异有统计学意义(P0.05);免疫荧光法测得Col-Ⅰ平均水平分别为:45.581±6.378,33.110±1.161,23.327±0.784,15.731±2.243,27.331±2.430,33.592±2.684,除10-6 mol/L组外任意组与空白对照组比较,差异有统计学意义(P0.05);ELISA法测得OCN平均水平分别为:3.002±1.639,1.711±0.370,2.193±0.551,2.439±1.469,2.703±1.175,8.021±3.036,任意组与空白对照组比较,差异有统计学意义(P0.05)。结论血清素对成骨细胞成负性效应,这种负效应与血清素浓度呈剂量相关性,随其浓度增高呈先强后弱关系。
[Abstract]:Objective to study the dose correlation between serotonin and osteoblasts of SD rats. Methods 1) adherent culture and isolation of neonatal rat cranial bone cells. 10 ~ (-5) mol / L ~ (-6) mol / L ~ 10 ~ (-7) mol / L ~ (10 ~ (-8) mol / L ~ (10 ~ (-9) mol 路L ~ (-1)) serotonin) on day 5 (MTT) was used to analyze the relationship between intervention factors and proliferation. Analysis of the relationship between intervention factors and differentiation. (5) on the 15th day, the osteocalcin (Osteocalcinia) level was measured by ELISA method. The relationship between intervention factors and differentiation was analyzed. Statistical method was used to collect and analyze data by SPSS 20.0. ) means that univariate ANOVA was used for the comparison between different groups. Results the mean optical density of 10 ~ (-5) mol 路L ~ (-1) 10 ~ (-6) mol 路L ~ (-1) 10 ~ (-7) mol / L ~ (10 ~ (-7)) mol / L ~ (10 ~ (-8)) mol / L ~ (10 ~ (-9) mol / L ~ (10 ~ (-9))) was 0. 400 卤0. 028 0. 379 卤0. 023 0. 379 卤0. 047 0. 401 卤0. 038 ~ 0. 420 卤0. 038 ~ (18 卤0. 064), respectively. The average level of ALP measured by microenzyme mapping method was 0.046 卤0.002n 0.017 卤0.002n 0.026 卤0.002n 0.046 卤0.002n 0.051 卤0.002n 0.073 卤0.003 respectively, which was compared with that of the control group. The average level of Col- 鈪，

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