MyD88抑制肽对LPS诱导BV2小胶质细胞极化状态的抑制作用及其机制

doi:10.13481/j.1671-587x.20200501

吉林大学学报(医学版) ›› 2020, Vol. 46 ›› Issue (05): 899-904.doi: 10.13481/j.1671-587x.20200501

• 基础研究 •

MyD88抑制肽对LPS诱导BV2小胶质细胞极化状态的抑制作用及其机制

杨吉平¹, 费琳², 柴学军¹, 苟兴春¹

1. 西安医学院基础医学研究所陕西省缺血性心血管疾病重点实验室陕西省脑疾病防治重点实验室, 陕西西安 710021;
2. 西安交通大学第一附属医院精神心理科, 陕西西安 710061

收稿日期:2020-02-06 发布日期:2020-10-23
通讯作者: 苟兴春,教授,硕士研究生导师(Tel:029-86177556,E-mail:jeepyang@xiyi.edu.cn) E-mail:jeepyang@xiyi.edu.cn
作者简介:杨吉平(1980-),男,陕西省西安市人,副教授,医学博士,主要从事脑缺血及神经保护方面的研究。
基金资助:
国家自然科学基金资助课题（81971330）；陕西省教育厅重点科学研究计划资助课题（19JS061）；西安医学院国家基金培育项目资助课题（2017GJFY22）；西安医学院配套基金资助课题（2018PT07）

Inhibitory effect of MyD88 inhibitory peptide on polarization of BV2 microglial cells induced by LPS and its mechanism

YANG Jiping¹, FEI Lin², CHAI Xuejun¹, GOU Xingchun¹

1. Institute of Basic Medical Sciences, Shaanxi Key Laboratory of Ischemic Cardiovascular Disease, Shaanxi Key Laboratory of Brain Disorders, Xi'an Medical University, Xi'an 710021, China;
2. Department of Psychiatry and Psychology, First Affiliated Hospital, Xi'an Jiaotong University, Xi'an 710061, China

Received:2020-02-06 Published:2020-10-23

摘要/Abstract

摘要： 目的：观察髓样分化因子88（MyD88）抑制肽（MIP）对脂多糖（LPS）激活BV2小胶质细胞极化的影响，阐明其作用机制。方法：将对数生长期的BV2小胶质细胞分为对照组（不进行处理）、LPS组（给予1 mg·L^-1 LPS处理）和不同剂量MIP+LPS组（分别给予25、50和100μmol·L^-1 MIP预处理1 h后，再加入1 mg·L^-1 LPS）。采用MTT法检测各组细胞存活率，实时荧光定量PCR（RT-qPCR）法检测各组BV2小胶质细胞中白细胞介素1β（IL-1β）、白细胞介素4（IL-4）、白细胞介素10（IL-10）和白细胞介素18（IL-18）mRNA表达水平，Western blotting法检测各组BV小胶质细胞中诱导型一氧化氮合酶（iNOS）和精氨酸酶1（Arg-1）蛋白表达水平。结果：LPS组BV2小胶质细胞体积变大，突起增多，呈阿米巴状；不同剂量MIP+LPS组小胶质BV2细胞活化率较LPS组明显减少（P<0.05或P<0.01）。MTT法检测，与对照组比较，LPS组细胞存活率明显降低（P<0.01）；与LPS组比较，不同剂量MIP+LPS组BV2小胶质细胞存活率明显提高（P<0.05或P<0.01）。RT-qPCR法和Western blotting法检测，与LPS组比较，不同剂量MIP+LPS组BV2小胶质细胞中IL-1β和IL-18 mRNA表达水平及iNOS蛋白表达水平均明显降低（P<0.05或P<0.01），而IL-4和IL-10 mRNA表达水平及Arg-1蛋白表达水平明显升高（P<0.05或P<0.01），且呈剂量依赖性。结论：MIP能够抑制活化的BV2小胶质细胞向M1型极化，促进其向M2型转化，抑制炎症的过度激活。

关键词: 髓样分化因子88抑制肽, BV2小胶质细胞, 极化, 脂多糖

Abstract: Objective: To observe the effects of myeloid differentiation factor 88 (MyD88) inhibitory peptide (MIP) on the polarization of BV2 microglial cells activated by lipopolysaccharide (LPS), and to clarify its mechnism. Methods: The BV2 microglia in logarithmic growth stage were divided into control group (without treatment), LPS group (treated with 1 mg·L^-1 LPS) and different doses of MIP+LPS groups (25, 50 and 100 mol·L^-1 MIP were respectively administrated for 1 h, and then LPS was added). The cell survival rates in each group were determined by MTT assay. The expression levels of interleukin-1β(IL-1β), interleukin-4(IL-4), interleukin-10(IL-10) and interleukin-18(IL-18) mRNA in the BV2 microglial cells in various groups were detected by real-time fluorescent quantitative PCR(RT-qPCR), and the expression levels of inducible nitric oxide synthase (iNOS) and argininase-1 (Arg-1) proteins in the BV2 microglia in various groups were detected by Western blotting method. Results: The BV2 microglial cells in LPS group became larger in size and had more projections, which were amoeboid in shape. The activation rates of BV2 microglial cells in different doses of MIP+LPS groups were decreased significantly compared with LPS group(P<0.05 or P<0.01). The MTT results showed that compared with control group, the cell survival rates in LPS group was significantly decreased (P<0.01); compared with LPS group, the survival rates of BV2 microglial cells in different doses of MIP+LPS groups were significantly increased (P<0.05 or P<0.01). The RT-qPCR and Western blotting results showed that compared with LPS group, the expression levels of IL-1β mRNA, IL-18 mRNA, and iNOS protein in the BV2 microglial cells in different doses of MIP+LPS groups were significantly decreased (P<0.05 or P<0.01), while the expression levels of IL-4 mRNA, IL-10 mRNA and Arg-1 protein were significantly increased (P<0.05 or P<0.01) in a dose-dependent manner. Conclusion: MIP can inhibit the M1 polarization of activated BV2 microglial cells, promote the M2 phenotype transformation, and inhibit the over-activation of inflammation.

Key words: myeloid differentiation factor 88 inhibitory peptide, BV2 microglial cells, polarization, lipopolysaccharide

中图分类号:

R392.5

杨吉平, 费琳, 柴学军, 苟兴春. MyD88抑制肽对LPS诱导BV2小胶质细胞极化状态的抑制作用及其机制[J]. 吉林大学学报(医学版), 2020, 46(05): 899-904.

YANG Jiping, FEI Lin, CHAI Xuejun, GOU Xingchun. Inhibitory effect of MyD88 inhibitory peptide on polarization of BV2 microglial cells induced by LPS and its mechanism[J]. Journal of Jilin University(Medicine Edition), 2020, 46(05): 899-904.

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MyD88抑制肽对LPS诱导BV2小胶质细胞极化状态的抑制作用及其机制

Inhibitory effect of MyD88 inhibitory peptide on polarization of BV2 microglial cells induced by LPS and its mechanism

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