中国免疫学杂志2024,Vol.40Issue(5) :1036-1041.DOI:10.3969/j.issn.1000-484X.2024.05.020

BCG感染巨噬细胞对肾小管上皮细胞损伤和修复的影响

Effects of BCG-infected macrophages on renal tubular epithelial cell injury and repair

乔春林 吴子毅 孙湛 勾璇 王新敏 章乐
中国免疫学杂志2024,Vol.40Issue(5) :1036-1041.DOI:10.3969/j.issn.1000-484X.2024.05.020

BCG感染巨噬细胞对肾小管上皮细胞损伤和修复的影响

Effects of BCG-infected macrophages on renal tubular epithelial cell injury and repair

乔春林 1吴子毅 2孙湛 3勾璇 4王新敏 5章乐4
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作者信息

  • 1. 石河子大学医学院第一附属医院,石河子 832002;石河子大学医学院,石河子 832002;新疆地方与民族高发病教育部重点实验室,石河子 832002;国家卫生健康委中亚高发病防治重点实验室,石河子 832002
  • 2. 石河子大学医学院第一附属医院,石河子 832002;石河子大学医学院,石河子 832002;国家卫生健康委中亚高发病防治重点实验室,石河子 832002
  • 3. 新疆医科大学,乌鲁木齐 830001
  • 4. 石河子大学医学院,石河子 832002;新疆地方与民族高发病教育部重点实验室,石河子 832002;国家卫生健康委中亚高发病防治重点实验室,石河子 832002
  • 5. 石河子大学医学院第一附属医院,石河子 832002;新疆地方与民族高发病教育部重点实验室,石河子 832002;国家卫生健康委中亚高发病防治重点实验室,石河子 832002
  • 折叠

摘要

目的:探讨肾结核发生过程中结核分枝杆菌BCG 感染巨噬细胞对肾小管上皮细胞损伤和修复的影响.方法:Transwell建立BCG感染的M0型巨噬细胞(上室)与HK-2细胞(下室)共培养模型,100 ng/ml佛波酯(PMA)诱导THP-1人单核巨噬细胞24 h成为M0型巨噬细胞,建立BCG感染细胞模型,分别于感染12 h和24 h收集细胞总蛋白,Western blot检测M1型巨噬细胞标志物CD86及M2型巨噬细胞标志物CD206蛋白表达,ELISA检测细胞培养上清中M1型巨噬细胞极化标志因子IL-6和TNF-α表达、M2型巨噬细胞极化标志因子TGF-β表达;实验分为HK-2组、BCG+HK-2组、BCG+M0+HK-2组及M0+HK-2组,CCK-8检测各组HK-2细胞活力,Hoechst实验检测各组HK-2细胞凋亡,蛋白免疫印迹检测各组HK-2细胞上皮细胞标志物E-cadhren及成纤维细胞标志物 α-SMA等转分化指标表达.结果:BCG感染M0型巨噬细胞后,M1型巨噬细胞活力12 h高于24 h(P<0.05),M2型巨噬细胞活力24 h高于12 h(P<0.05);BCG感染的巨噬细胞与HK-2细胞共培养后,HK-2细胞活力24 h高于12 h(P<0.001),凋亡水平12 h高于24 h,上皮细胞标志蛋白E-cadherin蛋白水平24 h高于12 h(P<0.001),成纤维细胞标志蛋白α-SMA蛋白水平12 h高于24 h(P<0.01).结论:肾结核发生发展过程中,早期BCG感染巨噬细胞可能通过M1型极化促进肾小管上皮细胞炎症损伤;随着感染时间延长,可能通过M2型极化修复肾小管上皮细胞损伤,发挥保护作用.

Abstract

Objective:To investigate effect of Mycobacterium tuberculosis BCG-infected macrophages on damage and repair of renal tubular epithelial cells during development of renal tuberculosis.Methods:A co-culture model of BCG-infected M0 macrophages(upper chamber)and HK-2 cells(lower chamber)was established by Transwell,and THP-1 human monocyte macrophages were induced by 100 ng/ml phorbol ester(PMA)24 h to become M0 macrophages,and BCG infection cell model was established.Total cell protein was collected at 12 h and 24 h of infection,respectively.Western blot was used to detect expressions of M1 macrophage marker CD86 and M2 macrophage marker CD206 protein.M1 macrophage polarization marker cytokines IL-6 and TNF-α and M2 macrophage polarization marker cytokine TGF-β expressions in cell culture supernatant were detected by ELISA;experiment was divided into HK-2 group,BCG+HK-2 group,BCG+M0+HK-2 group and M0+HK-2 group,CCK-8 was used to detect viability of HK-2 cells in each group,and Hoechst test was used to detect HK-2 cells apoptosis in each group.Epithelial cell marker E-cadhren and fibroblast markerα-SMA expressions in HK-2 cells of each group were detected by Western blot.Results:After BCG infection of M0 macrophages,M1 macrophage viability was higher than 24 h at 12 h(P<0.05),and M2 macrophage was higher than 12 h at 24 h(P<0.05).After two cells co-culture,HK-2 cell viability was higher than 12 h at 24 h(P<0.001),apoptosis level was higher than 24 h at 12 h,epithelial cell marker protein E-cadherin protein level was higher than 12 h at 24 h(P<0.001),fibroblast level of cell marker protein α-SMA protein at 12 h was higher than that at 24 h(P<0.01).Conclusion:During development of renal tuberculosis,early BCG-infected macrophages may promote inflammatory injury of renal tubular epithelial cells through M1-type polarization;with prolongation of infec-tion time,they may repair renal tubular epithelial cells through M2-type polarization and plays an important protective role.

关键词

肾小管上皮细胞/巨噬细胞/极化/BCG/损伤

Key words

Tubular epithelial cells/Macrophage/Polarization/BCG/Injury

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基金项目

新疆地方与民族高发病教育部重点实验室开放基金(KF2021-5)

石河子大学科研项目(ZZZC20261A)

石河子大学医学院第一附属医院重点基金(ZD202006)

兵团指导性科技计划立项项目(2022)(2022ZD045)

兵团指导性科技计划立项项目(2022)(2022ZD073)

出版年

2024
中国免疫学杂志
中国免疫学会,吉林省医学期刊社

中国免疫学杂志

CSTPCDCSCD北大核心
影响因子:0.926
ISSN:1000-484X
参考文献量24
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