曲古抑菌素A抑制小鼠周围淋巴结细胞分泌功能  

Inhibitory role of trichostatin A in secretion function of mouse peripheral lymph node cells

作　　者：刘一佳 余静[2] 张伟 刘可 梁华平[2] 李遂焰[1] 严军[2] LIU Yijia;YU Jing;ZHANG Wei;LIU Keh;LIANG Huaping;LI Suiyan;YAN Jun(School of Life Science and Engineering,Southwest Jiaotong University,Chengdu,Sichuan Province,610031;State Key Laboratory of Trauma,Bums and Combined Injury,Research Institute of Surgery,Daping Hospital,Army Medical University(Third Military Medical University),Chongqing,400042;College of Emergency and Trauma,Hainan Medical University,Haikou,Hainan Province,570100,China)

机构地区：[1]西南交通大学生命科学与工程学院,成都610031 [2]陆军军医大学(第三军医大学)大坪医院野战外科研究部,创伤、烧伤与复合伤国家重点实验室重庆400042 [3]海南医学院急诊创伤学院,海口570100

出　　处：《第三军医大学学报》2020年第3期253-258,共6页Journal of Third Military Medical University

基　　金：国家自然科学基金面上项目(81471863);重庆市基础研究与前沿探索项目(CSTC2016jcyjA0048);重庆市自然科学基金(CSTC2018jcyjAX0258)~~

摘　　要：目的研究曲古抑菌素A(trichostatin A,TSA)对小鼠周围淋巴结细胞分泌功能的影响,并初步探讨其机制。方法颈椎脱臼法处死成年雄性C57BL/6小鼠,取周围淋巴结(peripheral lymph nodes, LN)、肠系膜淋巴结(mesenteric lymph nodes, MLN)、派伊尔结(Peyer’s patches, PP)、脾脏(spleen, SP)、骨髓(bone marrow, BM)和胸腺(thymus, Thy),采用qPCR检测各免疫器官中Musculin(MSC)、IL-17A和IL-22的mRNA水平。取小鼠高表达MSC、IL-17和IL-22的免疫组织制备细胞悬液,采用完全随机试验设计分为对照组、1μg/mL脂多糖(lipopolysaccharides, LPS)组、1μg/mL LPS+10 ng/mL TSA组、1μg/mL LPS+30 ng/mL TSA组和1μg/mL LPS+50 ng/mL TSA组,置于37℃、5%CO2培养箱中分别培养6、12 h和24 h。收集上清及细胞,采用ELISA检测细胞上清中TNF-α和IL-10的分泌水平,确定TSA处理的最佳时间点。采用qPCR检测该时间点细胞MSC、IL-17A和IL-22的mRNA水平。结果与野生型小鼠中枢免疫器官BM相比,小鼠LN高表达MSC、IL-17A和IL-22(P<0.01,P<0.05);同样与Thy相比,小鼠LN也高表达MSC、IL-17A和IL-22(P<0.05)。与LPS组相比,不同浓度TSA能够降低各时间点小鼠LN细胞分泌TNF-α和IL-10的水平(P<0.01,P<0.05),并下调IL-17A和IL-22的表达(P<0.01),同时上调MSC(P<0.01,P<0.05),其最佳时间点是12 h。结论 TSA能够抑制LPS刺激LN细胞的分泌功能,其机制可能与MSC、IL-17A和IL-22的表达变化有关。Objective To investigate the role of trichostatin A(TSA) and its mechanism on secretory function of peripheral lymph node cells in mice. Methods The adult male C57BL/6 mice were sacrificed and their peripheral lymph nodes(LN), mesenteric lymph nodes(MLN), Peyer’s patches(PP), spleen(SP), bone marrow(BM) and thymus(Thy) were collected. The mRNA levels of musculin(MSC), interleukin-17A(IL-17A) and interleukin-22(IL-22) in above immune organs were detected by real-time quantity polymerase chain reaction(qPCR). Then the cell suspensions with high mRNA levels of MSC, IL-17A and IL-22, were prepared and randomly divided into 6 groups, including control group, 1 μg/mL lipopolysaccharide(LPS) group, 1 μg/mL LPS+10 ng/mL TSA group, 1 μg/mL LPS +30 ng/mL TSA group and 1 μg/mL LPS + 50 ng/mL TSA group. After the above cells were cultured under 37 ℃ and 5% CO2 for 6, 12 or 24 h, the supernatant and cells were collected. The secretion levels of tumor necrosis factor-α(TNF-α) and interleukin-10(IL-10) from the supernatant were measured by enzyme linked immunosorbent assay(ELISA), while the optimal time point of TSA treatment was determined. And then, the mRNA levels of MSC, IL-17A and IL-22 were detected by qPCR at this time point. Results Our results suggested that the mouse LN had highly expressed MSC, IL-17A, and IL-22 when compared with BM(P<0.01, P<0.05), and similar results were seen when compared with Thy(P<0.05). Compared with the LPS group, different concentrations of TSA decreased the secretion levels of TNF-α and IL-10(P<0.01, P<0.05), and at the same time, the treatment down-regulated the expression levels of IL-17A and IL-22(P<0.01), while up-regulated that of MSC(P<0.01, P<0.05). The best time point was identified as 12 h. Conclusion TSA inhibits the secretory function of LN cells stimulated by LPS, which might be related with the changes of MSC, IL-17A and IL-22 expression.

关 键 词：曲古抑菌素A 周围淋巴结 musculin 白细胞介素-17A 白细胞介素-22 

分 类 号：R364.5[医药卫生—病理学] R967[医药卫生—基础医学]