A151对糖氧剥夺和脂多糖诱导的BV-2细胞极化的影响

doi:10.6040/j.issn.1671-7554.0.2020.1499

摘要/Abstract

摘要： 目的探讨抑制性寡核苷酸A151治疗对糖氧剥夺(OGD)和脂多糖(LPS)诱导的小胶质细胞M1/M2极化的影响。方法倒置显微镜下观察A151治疗对BV-2细胞形态改变的影响。ELISA检测细胞上清液中细胞因子含量。RT-PCR检测细胞因子转录水平。Western blotting检测小胶质细胞M1表型、M2表型表面标志物的表达,检测NLRP3炎症小体的表达。免疫荧光检测小胶质细胞M1表型、M2表型表面标志物的表达。结果 A151可抑制LPS和OGD共同刺激引起的BV-2细胞形态变化。A151能下调小胶质细胞M1表型表面标志物和高表达的细胞因子诱导型一氧化氮合酶(iNOS)、CD16/CD32、肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)(P均<0.001),下调TNF-α mRNA、IL-1β mRNA的转录水平(P均<0.001)。上调小胶质细胞M2表型表面标志物和高表达的细胞因子: 精氨酸酶-1(Arg-1)、CD206、白介素-10(IL-10)、白介素-4(IL-4)(P_Arg-1<0.001,P_CD206<0.001,P_IL-10<0.001,P_IL-4=0.046),上调IL-10 mRNA、IL-4 mRNA转录水平(P均<0.001),下调NLRP3炎症小体表达(P均<0.001)。结论 A151通过抑制NLRP3炎症小体的活化,促使小胶质细胞从M1表型向M2表型的极化,发挥调节炎症的作用。

关键词: 抑制性寡核苷酸A151, 小胶质细胞, 极化, 炎症反应, NLRP3炎症小体

Abstract: Objective To investigate the effects of inhibitory oligodeoxynucleotide A151 on M1/M2 polarization of microglia by glucose and oxygen deprivation(OGD)and lipopolysaccharide(LPS). Methods The morphological changes of BV-2 cells treated with A151 were observed with inverted microscope. The cytokine content in supernatant and cytokine transcription levels were detected with ELISA and RT-PCR, respectively. The expressions of M1 and M2 surface markers in microglia and NLRP3 inflammasome were determined with Western blotting and immunofluorescence. Results A151 inhibited the morphological changes of BV-2 cells induced by LPS and OGD; downregulated the surface markers and highly-expressed cytokines of M1 microglia: inducible nitric oxide synthase(iNOS)、CD16/CD32、tumor necrosis factor-α(TNF-α)、interleukin-1β(IL-1β)(P<0.001); downregulated the transcription levels of TNF-α mRNA and IL-1β mRNA(P<0.001); upregulated the surface markers of M2 microglia and highly-expressed cytokines: arginase-1(Arg-1)、CD206、interleukin-10(IL-10)、 interleukin-4(IL-4)(P_Arg-1<0.001, P_CD206<0.001, P_IL-10<0.001, P_IL-4=0.046); upregulated IL-10 mRNA and IL-4 mRNA transcription levels(P<0.001); down-regulated NLRP3 inflammasome expression(P<0.001). Conclusion By inhibiting the activation of NLRP3 inflammasome, A151 promotes the polarization of microglia induced by LPS and OGD from the pro-inflammatory M1 phenotype to the anti-inflammatory M2 phenotype, and plays a role in regulating inflammation.

Key words: Inhibitory oligodeoxynucleotide A151, Microglia, Polarization, Inflammatory response, NLRP3 inflammasome

中图分类号:

R743.3

闵傲雪,朱天瑞,张凤,王冉冉,李晓红. A151对糖氧剥夺和脂多糖诱导的BV-2细胞极化的影响[J]. 山东大学学报 (医学版), 2021, 59(3): 1-9.

MIN Aoxue, ZHU Tianrui, ZHANG Feng, WANG Ranran, LI Xiaohong. Effects of A151 on the polarization of BV-2 cells induced by glucose and oxygen deprivation and lipopolysaccharide[J]. Journal of Shandong University (Health Sciences), 2021, 59(3): 1-9.

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