模拟微重力下柚皮苷对T细胞共育的成骨细胞增殖分化的影响
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篇名: 模拟微重力下柚皮苷对T细胞共育的成骨细胞增殖分化的影响
TITLE: Effects of naringin on the proliferation and differentiation of osteoblasts co -cultured with T cells under simulated microgravity
摘要: 目的 在模拟微重力情况下,探索柚皮苷作用于T细胞-成骨细胞共育体系后,对成骨细胞增殖分化的影响。方法本实验设正常重力对照组、正常重力柚皮苷组、模拟微重力对照组、模拟微重力柚皮苷组,其中对照组分别在正常重力或模拟微重力条件下将T细胞与成骨细胞共育,柚皮苷组分别在对照组基础上加入1×10-5mol/L柚皮苷溶液。显微镜下观察成骨细胞形态学,CCK-8法检测成骨细胞增殖率,碱性磷酸酶(ALP)试剂盒检测ALP活性,实时荧光定量聚合酶链式反应检测Runt相关转录因子2(Runx2)、白细胞介素6(IL-6)mRNA相对表达水平,Westernblot法检测Runx2和IL-6蛋白相对表达水平。结果与正常重力对照组比较,模拟微重力对照组成骨细胞密度降低,仅有少量聚集,细胞形态以圆形居多;与模拟微重力对照组比较,模拟微重力柚皮苷组成骨细胞成梭形或多角形,形态饱满,成鱼群状聚集。与正常重力对照组比较,正常重力柚皮苷组成骨细胞增殖率、ALP活性、Runx2mRNA和蛋白相对表达水平显著升高,IL-6mRNA和蛋白相对表达水平显著降低(P<0.05);模拟微重力对照组成骨细胞增殖率、ALP活性、Runx2mRNA和蛋白相对表达水平显著降低,IL-6mRNA和蛋白相对表达水平显著升高(P<0.05)。与模拟微重力对照组比较,模拟微重力柚皮苷组成骨细胞增殖率、ALP活性、Runx2mRNA和蛋白相对表达水平显著升高,IL-6mRNA和蛋白相对表达水平显著降低(P<0.05)。结论在T细胞-成骨细胞共育体系中,模拟微重力可以抑制成骨细胞的增殖分化能力;该条件下柚皮苷可以改善成骨细胞的增殖分化能力,其作用机制与IL-6水平的降低及Runx2水平的升高有关。
ABSTRACT: OBJECTIVE To explore the effects of naringin on the proliferation and differentiation of osteoblasts in T cell - osteoblast co -culture system under simulated microgravity . METHODS This experiment was setting normal gravity control group , normal gravity naringin group ,simulated microgravity control group and simulated microgravity naringin group .In the control group,T cells were co -cultured with osteoblasts under normal gravity or simulated microgravity conditions . In the naringin group , 1×10-5 mol/L naringin solution was added additionally on the basis of control group .The morphology of osteoblasts was observed under the microscope . The proliferation rate of osteoblasts was detected by CCK -8 method. The alkaline phosphatase (ALP)activity was detected by ALP kit . mRNA relative expressions of Runt related transcription factor 2(Runx2)and interleukin -6(IL-6)were detected by real -time fluorescence quantitative polymerase chain reaction . The relative expression of Runx 2 and IL -6 protein were detected by Western blot . RESULTS Compared with normal gravity control group ,the density of osteoblasts in the simulated microgravity control group decreased ,only a small number of cells were aggregated ,and the cell morphology was mostly round ; compared with the simulated microgravity control group ,the osteoblasts in the simulated microgravity naringin group formed spindle or polygonal ,plump in shape ,and clustered in groups of fish . Compared with normal gravity control group ,the proliferation rate of osteoblasts ,ALP activity ,relative expressions of Runx 2 mRNA and protein in the normal gravity naringin group were significantly increased ,while the relative expressions of IL -6 mRNA and protein were significantly decreased (P< 0.05);the proliferation rate of osteoblasts ,ALP activity ,relative expressions of Runx 2 mRNA and protein in the simulated microgravity control group were significantly decreased ,while relative expressions of IL -6 mRNA and protein were significantly increased(P<0.05). Compared with the simulated microgravity control group ,the proliferation rate of osteoblasts ,ALP activity , relative expressions of Runx 2 mRNA and protein in the simulated microgravity naringin group were significantly increased ,while relative expressions of IL -6 mRNA and protein were significantly decreased (P<0.05). CONCLUSIONS In the T cell -osteoblast co-culture system ,simulated microgravity can inhibit the proliferation and differentiation of osteoblasts ;naringin can improve the proliferation and differentiation ability of osteoblasts under this condition, and its mechanism of action is related to the decrease of IL -6 level and the increase of Runx 2 level.
期刊: 2022年第33卷第19期
作者: 陈建兴,尹文哲,孙奇峰,周森,张亚龙,王翌
AUTHORS:
关键字: 柚皮苷;成骨细胞;模拟微重力;增殖分化;T细胞
KEYWORDS: naringin;osteoblasts;simulated microgravity ;
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