模拟微重力对小鼠肝kupffer细胞增殖及相关基因表达的影响  

Simulated microgravity affects the proliferation and related genes expression in murine liver Kupffer cells

作　　者：田西朋 孙宏伟[2] 周金莲[3] 李雨霏[2] 闫洪锋 徐冰心[4] 董满库[2] 张宏文[2] 崔彦[2] 

机构地区：[1]安徽医科大学解放军306临床学院,北京100101 [2]解放军第306医院普通外科 [3]解放军第306医院病理科 [4]解放军第306医院特种病科

出　　处：《中华肝胆外科杂志》2016年第8期557-561,共5页Chinese Journal of Hepatobiliary Surgery

基　　金：全军医学科研“十二五”重点项目（BWS11J051）;全军试验技术研究计划重点项目（SYFD1500128）

摘　　要：目的探讨旋转式细胞组织培养系统（RCCS）模拟微重力对小鼠肝Kupffer细胞增殖及相关基因表达的影响。方法应用RCCS建立模拟微重力细胞培养系统。将小鼠肝原代Kupffer细胞随机分为模拟微重力组（SMG）和正常重力组（NG）。分别于培养第3、5、7天收获细胞。采用血球计数板进行细胞计数，流式细胞仪检测细胞周期，实时荧光定量PCR测定PCNA、Ki．67、ERK、CDK2以及CyclinB基因表达水平。结果培养第3天时，SMG组细胞计数明显低于NG组（2．6±0．1比3．1±0．2，P〈0．05），培养第5、7天SMG组明显高于NG组（6．9±0．4比5．9±0．2，P〈0．05；8．4±0．3比6．5±0．3，P〈0．05）。流式细胞仪检测结果显示，培养第3天SMG组G0／G1期比例明显高于NG组（78．1±0．2比59．7±1．2，P〈0．05），S期、G2／M期比例明显低于NG组（12．0±0．4比27．6±0．9，P〈0．05；9．9±0．3比12．7±0．4，P〈0．05）；培养第5天，SMG组G0／G1期比例低于NG组（69．5±1．5比74．8±0．7，P〈0．05），S期、G2／M期比例高于NG组（21．2±1．5比17．3±0．4，P〈0．05；9．3±0．4比7．9±0．4，P〈0．05）；培养第7天，SMG组G0／G1期比例依然低于NG组（73．9±1．9比81．7±1．3，P〈0．05），S期、G2／M期比例亦高于NG组（18．9±1．9比12．1±0．8，P〈0．05；7．3±0．2比6．2±0．6，P〈0．05）。实时荧光定量PCR结果显示，与NG组相比，SMG组PCNA、Ki-67、ERK、CDK2以及CyclinB基因表达在培养3天时均下调，第5和7天呈现明显上调趋势。结论在RCCS模拟失重应激损伤期，小鼠肝Kupffer细胞增殖功能受到抑制，以后Kupffer细胞增殖能力在相关基因的参与调节下得以恢复并被激活强化。Objective To investigate the effects of simulated microgravity on the proliferation and expression of related genes in murine liver Kupffer cells. Methods The rotary cell culture system （RCCS） was used to simulate the microgravity environment and culture the murine liver Kupffer cells, which were divided into simulated microgravity group （SMG） and normal gravity group （NG） at random, and were harvested after being cultured for 3, 5 and 7 days, respectively. The cell count and cell cycle were detected by hemocytometer and flow cytometry. The Kupffer cell＇ s expression of PCNA, Ki-67, ERK, CDK2 and Cyclin B were assayed by real-time fluorescent quantitative PCR （qPCR）. Results After being cuhured for 3 days, the number of Kupffer cells in SMG group were less than that in NG group （2.6 ± 0.1 vs. 3.1 ± 0. 2, P 〈 0.05 ） , while after being cultured for 5 and 7 days, the number of Kupffer cells in SMG group were increased significantly compared with that in NG group （6.9 ± 0.4 vs. 5.9 ± 0.2, P 〈 0.05 ; 8.4 ± 0.3 vs. 6.5 ± 0. 3, P 〈 0. 05 ）. The flow cytometry also revealed that at 3rd days of culture, the G0/G1 phase of Kupffer cells in SMG group was significantly higher than that in NG group （78.1 ± 0.2 vs. 59.7 ± 1.2, P 〈 0.05 ）, while the S phase and the G2/M phase were significantly lower than that in NG group （ 12.0 ± 0.4 vs. 27.6 ±0.9, P〈0.05; 9.9 ±0.3 vs. 12.7 ±0.4, P〈0.05）. In contrary, the G0/G1 phase of Kupffer cells being cultured for 5 days in SMG group was decreased （ 69.5± 1.5 vs. 74.8 ± 0.7, P 〈 0.05 ）, and the S phase and G2/M phase were increased （21.2 ±1.5 vs. 17.3 ±0.4, P〈0.05; 9.3 ±0.4 vs. 7.9± 0.4, P 〈 0.05 ） significantly compared with those in NG group. After 7 days of culture, the G0/G1 phase of the SMG group was still decreased （73.9 ± 1.9 vs. 81.7 ± 1.3, P 〈 0.05 ） while the S phase and the G2/M phase were also increased （ 18.9 ± 1.9 vs. 12.1 ± 0.8, P 〈 0.05 ; 7.3 ± 0.2 vs. 6.2 ± 0.6） compared with

关 键 词：旋转式细胞组织培养系统 模拟微重力 肝脏 KUPFFER细胞 增殖 基因 

分 类 号：R852[医药卫生—航空、航天与航海医学]