miR-216a-5p在HMGB1介导的表皮葡萄球菌感染致腹膜透析相关腹膜炎中的作用
作者:
作者单位:

1.山东第一医科大学附属省立医院医院感染管理办公室, 山东 济南 250021;2.山东大学附属省立医院肾内科, 山东 济南 250021

作者简介:

通讯作者:

李卫光  E-mail: emlwg@sina.com

中图分类号:

R378.1;R446.5

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The role of miR-216a-5p in HMGB1-mediated peritoneal dialysis-associa-ted peritonitis induced by Staphylococcus epidermidis infection
Author:
Affiliation:

1.Department of Healthcare-associated Infection Mana- gement, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan 250021, China;2.Department of Nephrology, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250021, China

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    摘要:

    目的 探讨miR-216a-5p在高迁移率蛋白B1(HMGB1)介导的表皮葡萄球菌感染致小鼠腹膜透析相关腹膜炎(PDAP)中的作用及机制。 方法 选取健康雄性C57BL/6J小鼠,随机分为对照组、感染组、感染+HMGB1抑制剂组,收集腹腔积液及腹膜组织分别进行白细胞计数、HE和免疫组织染色;Real-time PCR和Western Blot检测白细胞介素-1α(IL-1α)、白细胞介素-6(IL-6)、肿瘤坏死因素-α(TNF-α)、HMGB1、核转录因子-κB(NF-κB)和核转录因子抑制蛋白(I-κB)的mRNA和蛋白表达水平;生信预测发现,miR-216a-5p可能与HMGB1结合,参与其诱导的PDAP的发生,并构建感染+miR-216a-5p mimics组小鼠,通过双荧光素酶报告基因检测验证miR-216a-5p和HMGB1的关系,采用Real-time PCR、Western Blot、免疫组织染色检测HMGB1的表达变化。 结果 与对照组相比,感染组小鼠腹腔积液白细胞计数增多,炎性浸润明显,IL-1α、IL-6、TNF-α,以及HMGB1 mRNA和蛋白表达均升高(均P < 0.05);HMGB1抑制剂(甘草素)干预后,小鼠腹腔积液白细胞计数下降,炎性浸润改善,IL-1α、IL-6、TNF-α,以及HMGB1 mRNA和蛋白表达均下降(均P < 0.05)。感染组小鼠NF-κB表达高于对照组,I-κB表达低于对照组;HMGB1抑制剂干预后NF-κB表达降低,I-κB表达升高(均P < 0.05)。Real-time PCR结果证实,与对照组相比,miR-216a-5p的含量在感染组中显著减少;双荧光素酶报告基因检测提示,miR-216a-5p可直接作用于HMGB1的3’UTR区;与感染组相比,感染+miR-216a-5p mimics组小鼠中HMGB1 mRNA和蛋白表达均下降(均P < 0.05)。 结论 HMGB1在表皮葡萄球菌感染致小鼠PDAP中发挥重要作用,抑制HMGB1可改善小鼠炎症反应,miR-216a-5p可通过靶向HMGB1参与表皮葡萄球菌感染致PDAP的发生。

    Abstract:

    Objective To investigate the role and mechanism of miR-216a-5p in high mobility group protein-B1 (HMGB1)-mediated peritoneal dialysis-associated peritonitis (PDAP) induced by Staphylococcus epidermidis infection in mice. Methods Healthy male C57BL/6J mice were selected and randomly divided into control group, infection group, and infection+HMGB1 inhibitor group, peritoneal effusion and peritoneal tissue was collected for detection of whit blood cell (WBC) count, HE and immunohistochemical staining; mRNA and protein expression levels of interleukin-1α(IL-1α), interleukin-6 (IL-6), tumor necrosis factors-α(TNF-α), HMGB1, NF-κB, and I-κBi were detected by real-time polymerase chain reaction (RT-PCR) and Western Blot; bioinformatics prediction found that miR-216a-5p may bind with HMGB1 and participate in the occurrence of HMGB1-mediated PDAP, and construct dinfection+miR-216a-5p mimics mice group, relationship between miR-216a-5p and HMGB1 was verified by double luciferase reporter gene assay, changes in expression of HMGB1 were detected by RT-PCR, Western blot and immunohistochemical staining. Results Compared with control group, WBC count of peritoneal effusion in mice in infection group increased, inflammatory infiltration was obvious, expression of mRNA and protein of IL-1α, IL-6, TNF-α and HMGB1 all increased (all P < 0.05); after intervention by HMGB1 inhibitor (glycyrrhizin), WBC count of mice peritoneal effusion decreased, inflammatory infiltration improved, expression of mRNA and protein of IL-1α, IL-6, TNF-α and HMGB1 all decreased (all P < 0.05). The expression of NF-κB in mice in infection group was higher than that in control group, I-κBi was lower than that in control group; after HMGB1 inhibitor intervention, NF-κB expression decreased, I-κB expression increased (both P < 0.05). RT-PCR results confirmed that the content of miR-216a-5p decreased significantly in infection group compared with control group; dual-luciferase reporter gene assay suggested that miR-216a-5p could directly act on the 3' UTR region of HMGB1; compared with infection group, the expression of mRNA and protein of HMGB1 decreased in mice in infection+miR-216a-5p mimics group (both P < 0.05). Conclusion HMGB1 plays an important role in PDAP induced by Staphylococcus epidermidis infection, inhibiting HMGB1 can improve the inflammatory response in mice, miR-216a-5p can participate in the occurrence of PDAP induced by Staphylococcus epidermidis infection through targeting HMGB1.

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贾磊,刘冰,李卫光. miR-216a-5p在HMGB1介导的表皮葡萄球菌感染致腹膜透析相关腹膜炎中的作用[J]. 中国感染控制杂志,2021,(11):1028-1034. DOI:10.12138/j. issn.1671-9638.20218421.
Lei JIA, Bing LIU, Wei-guang LI. The role of miR-216a-5p in HMGB1-mediated peritoneal dialysis-associa-ted peritonitis induced by Staphylococcus epidermidis infection[J]. Chin J Infect Control, 2021,(11):1028-1034. DOI:10.12138/j. issn.1671-9638.20218421.

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  • 收稿日期:2021-05-20
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  • 在线发布日期: 2024-04-26
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