Department of Respiratory Medicine, 306th Hospital of PLA, 9 North Anxiang Road, Chaoyang District, Beijing, 100101, China.
Department of Neurology, 306th Hospital of PLA, Beijing, China.
Inflammation. 2017 Apr;40(2):589-600. doi: 10.1007/s10753-016-0506-0.
We evaluated different lymphocyte populations and levels of plasma cytokines in peripheral blood as well as inflammatory infiltration and expressions of cytokines in lung tissues derived from macaque under long-term stimulated microgravity through being suspended in an antiorthostatic position so as to identify relevant immune parameters and to understand potential mechanisms of lung injury. Fifteen healthy male rhesus macaques were randomly divided into groups 1 (control, n = 5), groups 2 (head-down tilting for 6 weeks, n = 5), and groups 3 (head-down tilting for 6 weeks and recovery for 4 weeks, n = 5). Lymphocyte subsets in peripheral blood were analyzed using flow cytometry and the concentrations of 14 cytokines in plasma were measured with Luminex multiplexing technology. HE staining and transmission electron microscopy were employed to investigate the morphologies and subcellular structures of lung tissues. Immunohistochemistry and real-time PCR were employed to explore mRNA and protein expressions of cytokines in lung tissues. Immunohistochemical demonstrations were detected for CD3, CD4, CD8 T lymphocytes, CD20 B lymphocytes, and CD68 macrophages in lung tissues. Compared to group 1, groups 2 and 3 showed a decrease in the percentage of CD2+T cells, CD2+CD4+T helper cells, and CD2+CD8+cytotoxic T cells as well as an increase in the expression of CD95 on the surface of T lymphocytes in peripheral blood. The serum cytokine levels of IL-18 and TNF-α were increased in group 2 when compared to groups 1 and 3. HE and TEM observed changes in the structure and ultrastructure of lung tissues in groups 2 and 3. The number of CD3+T cell, CD4+T cell, CD8+T cells, and CD68+macrophage and the expression levels of IL-1β, IL-6, and IL-18 in lung tissues were increased in groups 2 when compared with groups 1 and 3. Our data suggested that long-term microgravity might alter the functions of immune system and cause lung damage, changing lymphocyte distribution and functions as well as cytokine production.
我们评估了猕猴在长期模拟微重力下被悬吊于反姿势位后外周血中的不同淋巴细胞群和血浆细胞因子水平以及肺部的炎症浸润和细胞因子表达,以确定相关的免疫参数,并了解肺部损伤的潜在机制。15 只健康雄性恒河猴随机分为 3 组:对照组(n=5)、头低位倾斜 6 周组(n=5)和头低位倾斜 6 周+恢复 4 周组(n=5)。使用流式细胞术分析外周血中的淋巴细胞亚群,使用 Luminex 多重分析技术测量血浆中 14 种细胞因子的浓度。采用 HE 染色和透射电镜观察肺组织的形态和亚细胞结构。采用免疫组化和实时 PCR 检测肺组织中细胞因子的 mRNA 和蛋白表达。免疫组化检测肺组织中 CD3、CD4、CD8 T 淋巴细胞、CD20 B 淋巴细胞和 CD68 巨噬细胞。与第 1 组相比,第 2 组和第 3 组外周血中 CD2+T 细胞、CD2+CD4+辅助性 T 细胞和 CD2+CD8+细胞毒性 T 细胞的百分比下降,T 淋巴细胞表面 CD95 的表达增加。与第 1 组和第 3 组相比,第 2 组血清中白细胞介素 18 和肿瘤坏死因子-α的水平升高。HE 和 TEM 观察到第 2 组和第 3 组肺组织的结构和超微结构发生变化。与第 1 组和第 3 组相比,第 2 组肺组织中 CD3+T 细胞、CD4+T 细胞、CD8+T 细胞和 CD68+巨噬细胞的数量以及白细胞介素 1β、白细胞介素 6 和白细胞介素 18 的表达水平增加。我们的数据表明,长期微重力可能改变免疫系统的功能,导致肺部损伤,改变淋巴细胞分布和功能以及细胞因子的产生。