LncRNA GSEC/CTCF通过调节巨噬细胞极化促进布氏杆菌性脊柱炎的机制研究
LncRNA GSEC/CTCF promotes Brucella spondylitis by regulating macrophage polarization
王 兴 阿日奔吉日嘎拉 高 飞 刘志恒 刘雄伟 李斯琴
作者信息
摘要
目的 探讨LncRNA GSEC在布氏杆菌性脊柱炎（BS）中的作用，及其对BS患者巨噬细胞极化的调节机制。方法 检测BS患者和健康志愿者血浆中M1和M2巨噬细胞比例，以及LncRNA GSEC表达水平。以佛波酯诱导人单核细胞株THP1分化为巨噬细胞。对照组细胞培养在补充了5%健康体检者血清的RPMI-1640培养基中；BS组细胞培养在补充了5%的BS患者血清的RPMI-1640培养基中；Sh-NC组和sh-GSEC组采用含sh-NC或sh-GSEC的慢病毒载体转染THP1细胞，sh-GSEC+LV-NC组和sh-GSEC+LV-CTCF组采用含sh-GSEC的慢病毒载体与含LV-NC或LV-CTCF的慢病毒载体共转染THP1细胞，并将上述4组细胞均培养在补充了5%的BS患者血清的RPMI-1640培养基中。RIP分析LncRNA GSEC与CTCF的结合；ELISA检测血清和细胞上清液中IL-1β、IL-6、TNF-α、IL-4、IL-10和TGF-β水平；流式细胞术分析M1和M2巨噬细胞比例。结果 与对照组相比，BS组患者血浆M1巨噬细胞和血清炎症因子IL-1β、IL-6、TNF-α水平显著升高，M2巨噬细胞和血清抗炎症因子IL-4、IL-10、TGF-β水平显著降低（P<0.05）。LncRNA GSEC表达在BS患者血浆中显著升高（P<0.01）。敲低GSEC抑制巨噬细胞M1极化，促进M2极化，抑制炎症因子分泌（P<0.05）。此外，发现CTCF表达在BS患者血浆中显著上调，与LncRNA GSEC表达呈显著正相关（R2=0.913，P<0.001）。过表达CTCF逆转了LncRNA GSEC敲低对巨噬细胞极化的调节作用（P<0.05）。结论 LncRNA GSEC/CTCF可能通过诱导巨噬细胞M1极化，抑制M2极化，促进布氏杆菌性脊柱炎。
Abstract
Objective To investigate the role of LncRNA GSEC in Brucella spondylitis (BS) and its regulation effect on macrophage polarization in BS patients. Methods The proportions of M1 and M2 macrophages and the expression levels of LncRNA GSEC in the plasma of BS patients and healthy volunteers were detected. Human monocyte cells THP1 were induced to differentiate into macrophages by Phorbol myristate acetate (PMA). The cells of the control group were cultured in RPMI-1640 medium supplemented with 5% of the serum of healthy people; cells of the BS group were cultured in RPMI-1640 medium supplemented with 5% of the serum of BS patients; THP1 cells were transfected with lentiviral vectors containing sh-NC or sh-GSEC in sh-NC group and sh-GSEC group, and THP1 cells were co-transfected with lentiviral vectors containing sh-GSEC and lentiviral vectors containing LV-NC or LV-CTCF in sh-GSEC+LV-NC group and sh-GSEC+LV-CTCF group, and the above four groups of cells were cultured in RPMI-1640 medium supplemented with 5% of the serum of BS patients. RIP analysis was performed to verify the interaction between LncRNA GSEC and CTCF; The levels of IL-1, IL-6, TNF-, IL-4, IL-10 and TGF- in serum and cell supernatants were detected with ELISA; The percentages of M1 and M2 macrophages were analyzed by the flow cytometry. Results Compared with the control group, the levels of M1 macrophages in plasma and inflammatory factors IL-1, IL-6, TNF- in serum were significantly increased, while the levels of M2 macrophages in plasma and anti-inflammatory factors IL-4, IL-10, TGF- in serum of BS group were significantly decreased (P<0.05). The expression of LncRNA GSEC was significantly upregulated in the plasma of BS patients (P<0.01). Knockdown of LncRNA GSEC inhibited macrophage M1 polarization, promoted M2 polarization, and inhibited the secretion of inflammatory factors (P<0.05). In addition, we found that CTCF expression was significantly upregulated in the plasma of BS patients, and it was significantly positively correlated with LncRNA GSEC expression (R2=0.913, P<0.001). Overexpression of CTCF reversed the regulatory effect of LncRNA GSEC knockdown on macrophage polarization (P<0.05). Conclusion LncRNA GSEC/CTCF promotes Brucella spondylitis by inducing M1 polarization of macrophages and inhibiting M2 polarization.关键词
布氏杆菌性脊柱炎/LncRNA GSEC/CTCF/巨噬细胞极化Key words
Brucella spondylitis/LncRNA GSEC/CTCF/Macrophage polarization引用本文复制引用
王 兴,阿日奔吉日嘎拉,高 飞,刘志恒,刘雄伟,李斯琴.LncRNA GSEC/CTCF通过调节巨噬细胞极化促进布氏杆菌性脊柱炎的机制研究[EB/OL].(2024-09-30)[2026-05-19].https://chinaxiv.org/abs/202409.00259.学科分类
基础医学/临床医学/内科学
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