模拟微重力环境对小鼠成纤维细胞增殖和创伤修复相关蛋白基因表达的影响

Effect of Simulated Microgravity Environment on Fibroblasts Proliferation and Expressions of Wound Healing Related Proteins' Genes in Mice

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 investigate effect of simulated microgravity environment by rotary cell culture system （RCCS） on fibroblasts proliferation （L929 cells） and expression changes of wound healing related proteins＇ genes in mice. Methods L929 cells were randomly divided into microgravity group and normal gravity group, and were respec-tively cultured in RCCS for 3, 5 and 7 ds. Cells cycles were detected by flow cytometry, and then mRNA expressions of wound healing related proteins were detected using quantitative real-time polymerase chain reaction （ PCR） . Cell-micro-carrier suspensions of both groups were obtained after 7 ds of culture, and the changes of structure morphology were ob-served. Results Compared with those in normal gravity group, L929 cell numbers in G1 phase on the 3rd, 5th and 7th ds culture were significantly less in microgravity group;in S phase cell numbers were increased on the 3rd and 5th ds culture, but the number was decreased on the 7th d culture, while in G2/M phase cell numbers were increased on the 3rd and 7th ds culture, and the number was decreased on the 7thd culture in microgravity group （P〈0. 05）. Compared with those in normal gravity group, in microgravity group, mRNA expressions of Collagen Ⅰ （ ColⅠ） , Collagen III （ ColⅢ） and transforming growth factor-β（TGF-β） were lower on the 3rd d culture （P〈0. 05）;on the 5th d culture, Caspase-3 mR-NA expression was lower, while mRNA expressions of ColⅠ, TGF-βand Bcl-2 were higher in microgravity group （ P〈0. 05）;mRNA expressions of wound healing related proteins in 7ds were higher on the 7th d culture （P〈0. 05）. Simula-ted microgravity environment after 7 ds of culture affected microfilament structure of L929 cells, but there were no signifi-cant differences between two groups. Conclusion RCCS simulated microgravity environment may change cycle conver-sion of fibroblast L929 cells, enhance ability of cell proliferation and affect expressions of wound healing related proteins＇ genes in mice.