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目的研究微重力细胞培养系统(RCCS)模拟微重力环境下小鼠成纤维细胞(L929细胞)增殖及创伤修复相关蛋白基因表达的变化。方法将L929细胞随机分为微重力组和正常重力对照组,分别在RCCS中培养3、5、7天,流式细胞仪检测细胞周期,实时荧光定量聚合酶链反应检测创伤修复蛋白mRNA表达水平。培养第7天取两组细胞-微载体悬液观察细胞微丝结构形态变化。结果微重力组L929细胞的G1期细胞在第3、5、7天明显少于正常重力对照组(P<0.05);与正常重力对照组相比,微重力组S期细胞在第3、5天有所增加,第7天减少,G2/M期细胞在培养第3、7天升高,培养第5天下降(P<0.05)。培养第3天微重力组Ⅰ型胶原蛋白(ColⅠ)和Ⅲ型胶原蛋白(ColⅢ)、β转化生长因子(TGF-β)mRNA表达低于正常重力对照组(P<0.05);培养第5天微重力组Caspase-3 mRNA表达低于正常重力对照组,ColⅠ、TGF-β及Bcl-2 mRNA表达均高于正常重力对照组(P<0.05);培养第7天微重力组创伤修复相关蛋白mRNA表达均高于正常重力对照组(P<0.05)。模拟微重力环境下培养7 d对L929细胞微丝结构有影响但两组间差异无明显差别。结论模拟微重力环境能改变L929细胞周期转化、增强细胞增殖能力,并影响创伤修复相关蛋白的表达。
Objective To study the changes of proliferation and expression of wound healing related gene in murine fibroblasts (L929 cells) under the condition of microgravity cell culture system (RCCS) simulated microgravity. Methods L929 cells were randomly divided into microgravity group and normal gravity control group. The cells were cultured in RCCS for 3, 5 and 7 days respectively. The cell cycle was detected by flow cytometry. The mRNA expression level of wound repair protein was detected by real-time fluorescence quantitative polymerase chain reaction . On the 7th day after culture, the morphological changes of microfilaments in the two groups of cells-microcarrier suspension were observed. Results In the microgravity group, the number of cells in G1 phase of L929 cells was significantly less than that of the normal control group on the 3rd, 5th and 7th day (P <0.05). Compared with the normal control group, Day and decreased on the 7th day. G2 / M phase cells increased on the 3rd and 7th day and decreased on the 5th day (P <0.05). On day 3, the mRNA expression of type Ⅰ collagen and type Ⅲ collagen Ⅲ and TGF-β in microgravity group were lower than those in normal control group (P <0.05) The expression of Caspase-3 mRNA in the microgravity group was lower than that in the normal control group (P <0.05). The mRNA expression of ColⅠ, TGF-β and Bcl-2 in the microgravity group was significantly higher than that in the normal gravity group mRNA expression were higher than the normal gravity control group (P <0.05). Under the simulated microgravity environment, the culture of L929 cells for 7 days had an effect on the microfilament structure, but there was no significant difference between the two groups. Conclusion Simulated microgravity can change the cell cycle of L929 cells, enhance the cell proliferation and affect the expression of wound repair related proteins.