模拟微重力对人单核细胞白血病THP-1细胞脂多糖诱导的炎性细胞因子释放的影响

中华航空航天医学杂志 Pub Date : 2018-06-15 DOI:10.3760/CMA.J.ISSN.1007-6239.2018.02.008

Ying Zhang, Jing-Chan Yao, Tingting Li, Jieping Li, Jingyu Wang, Min Yuan, Jiang Cheng

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When the cells in simulated weightlessness group were cultured in the clinostat for 48 h, 3 bottles of cell for each group were centrifuged for 5 min with the speed of 1 000 r/min. Then the LPS cell was added to stimulate cells with the concentration of 1 μg/ml and the cultivation lasted for 8 h. The cells and the cultured medium were collected by centrifugation. The other 3 bottles of control group and simulated weightlessness group were added equivalent volume cultured medium with LPS. The cells and the cultured medium were collected by centrifugation 8 h later. The experiment was repeated for 3 times. 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引用次数: 1

摘要

目的观察模拟微重力对脂多糖(LPS)诱导的单核细胞白血病THP-1细胞炎性因子释放的影响。方法以人THP-1细胞为研究对象，分为对照组和模拟失重组。模拟失重组细胞复苏传代后，用恒温器以30 r/min的速度处理，在37℃培养箱中培养。对照组与模拟失重组同步设置，但不进行回转器处理。每组6瓶细胞检测3次。模拟失重组细胞在恒温器中培养48 h时，每组3瓶细胞以1 000 r/min的速度离心5 min。然后加入浓度为1 μg/ml的LPS细胞刺激细胞，培养8 h，离心收集细胞和培养基。另外3瓶对照组和模拟失重组分别添加等体积LPS培养基。8 h后离心收集细胞和培养基。实验重复3次。比较4种实验条件下收集的mRNA表达及TNF-α、IL-6、IL-1β浓度的变化。结果①与对照组比较，LPS组和模拟失重+ LPS组大鼠血清TNF-α、IL-6、IL-1β mRNA表达量显著升高(q=11.63 ~ 50.24, P<0.05);模拟失重组TNF-α、IL-1β mRNA表达量显著升高(q=5.56 ~ 3.44, P<0.05)。与LPS组比较，模拟失重+ LPS组TNF-α、IL-6、IL-1β mRNA表达量显著升高(q=9.08 ~ 26.50, P<0.01)。②与对照组比较，LPS组和模拟失重组小鼠血清TNF-α、IL-6、IL-1β浓度显著升高(q=3.02 ~ 32.52, P<0.05);模拟失重+ LPS组小鼠血清TNF-α、IL-6、IL-1β浓度显著升高(q=40.02 ~ 65.31, P<0.01)。与LPS组比较，模拟失重+ LPS组TNF-α、IL-6、IL-1β浓度显著升高(q=17.59 ~ 32.79, P<0.01)。结论模拟失重可显著增加THP-1细胞炎症因子的分泌，加重lps诱导的炎症反应。关键词:模拟失重;脂多糖;基因表达;细胞因子

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Effect of simulated microgravity on lipopolysaccharide induced inflammatory cytokines release in human monocytic leukemia THP-1 cells

Objective To observe the effect of simulated microgravity on lipopolysaccharide (LPS)-induced inflammatory cytokines release in human monocytic leukemia THP-1 cells. Methods The human′s THP-1 cells were chosen as the research objects and divided into control group and simulated weightlessness group. After being resuscitated and subcultured, the cells in simulated weightlessness group were treated by clinostat with the speed of 30 r/min and cultured in the 37℃ incubator. The control group was set up synchronously with the experimental conditions that were consistent with simulated weightlessness group but without clinostat treatment. Six bottles of cells for each group had been tested for 3 times. When the cells in simulated weightlessness group were cultured in the clinostat for 48 h, 3 bottles of cell for each group were centrifuged for 5 min with the speed of 1 000 r/min. Then the LPS cell was added to stimulate cells with the concentration of 1 μg/ml and the cultivation lasted for 8 h. The cells and the cultured medium were collected by centrifugation. The other 3 bottles of control group and simulated weightlessness group were added equivalent volume cultured medium with LPS. The cells and the cultured medium were collected by centrifugation 8 h later. The experiment was repeated for 3 times. The mRNA expression and the changes of the concentration of TNF-α, IL-6, IL-1β gathered under 4 experimental conditions were compared. Results ①Compared with those in the control group, the mRNA expressions of TNF-α, IL-6 and IL-1β were increased significantly in LPS group and simulated weightlessness + LPS group (q=11.63-50.24, P<0.05); mRNA expressions of TNF-α and IL-1β were increased significantly in simulated weightlessness group (q=5.56-3.44, P<0.05). Compared with LPS group, the mRNA expressions of TNF-α and IL-6, IL-1β were increased significantly in simulated weightlessness + LPS group (q=9.08-26.50, P<0.01). ②Compared with those in control group, the concentration of TNF-α, IL-6, IL-1β were increased significantly in LPS group and simulated weightlessness group (q=3.02-32.52, P<0.05); the concentrations of TNF-α, IL-6 and IL-1β were increased significantly in simulated weightlessness + LPS group (q=40.02-65.31, P<0.01). Compared with those in LPS group, the concentrations of TNF-α and IL-6, IL-1β were increased significantly in simulated weightlessness + LPS group (q=17.59-32.79, P<0.01). Conclusions Simulated weightlessness can significantly increase the secretion of inflammatory cytokines and aggravate the LPS-induced inflammatory reaction in THP-1 cells. Key words: Weightlessness simulated; Lipopolysaccharides; Gene expression; Cytokines

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中华航空航天医学杂志航空航天医学

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期刊介绍： The aim of Chinese Journal of Aerospace Medicine is to combine theory and practice, improve and popularize, actively advocate a hundred flowers bloom and a hundred schools of thought contend, advocate seeking truth from facts, promote the development of the related disciplines of aerospace medicine and human efficiency, and promote the exchange and penetration of aerospace medicine and human efficiency with other biomedical and engineering specialties. Topics of interest for Chinese Journal of Aerospace Medicine include: -The content of the journal belongs to the discipline of special medicine and military medicine, with the characteristics of multidisciplinary synthesis and cross-penetration, and mainly reflected in the aerospace industry, aerospace flight safety and efficiency, as well as the synthesis of special medicine, preventive medicine, environmental medicine, psychology, etc. -Military aeromedicine (Air Force, Navy and Army aeromedicine) and civil aeromedicine, with a balance of aerospace medicine are the strengths of the journal. -The change in aerospace medicine from a focus on promoting physiological compensatory adaptations to enhancing human performance under extreme environmental conditions is what the journal is helping to promote. -The expansion of manuscripts in high altitude medicine is also a special emphasis of the journal.