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

Abstract

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

CLC Number:

R392.5

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.

References

[1] HARUWAKA K, IKEGAMI A, TACHIBANA Y, et al. Dual microglia effects on blood brain barrier permeability induced by systemic inflammation[J]. Nat Commun, 2019, 10(1):5816.
[2] DI STADIO A, ANGELINI C. Microglia polarization by mitochondrial metabolism modulation:a therapeutic opportunity in neurodegenerative diseases[J]. Mitochondrion, 2019, 46:334-336.
[3] WANG J, XING H Y, WAN L, et al. Treatment targets for M2 microglia polarization in ischemic stroke[J]. Biomed Pharmacother, 2018, 105:518-525.
[4] FIEBICH B L, BATISTA C R A, SALIBA S W, et al. Role of microglia TLRs in neurodegeneration[J]. Front Cell Neurosci, 2018, 12:329.
[5] LIU M J, XIE J H, SUN Y X. TLR4/MyD88/NF-κB-mediated inflammation contributes to cardiac dysfunction in rats of PTSD[J]. Cell Mol Neurobiol, 2020, 40(6):1029-1035.
[6] YAN H Y, ZHANG D D, WEI Y X, et al. Inhibition of myeloid differentiation primary response protein 88 provides neuroprotection in early brain injury following experimental subarachnoid hemorrhage[J]. Sci Rep, 2017, 7(1):15797.
[7] YANG J P, ZHAO Y Y, ZHANG L, et al. RIPK3/MLKL-mediated neuronal necroptosis modulates the M1/M2 polarization of microglia/macrophages in the ischemic cortex[J]. Cereb Cortex, 2018, 28(7):2622-2635.
[8] KUČIĆ N, RAČKI V, JURDANA K, et al. Immunometabolic phenotype of BV-2 microglia cells upon murine cytomegalovirus infection[J]. J Neurovirol, 2019, 25(4):496-507.
[9] YUN H K, PARK J, CHAE U, et al. Parkin in early stage LPS-stimulated BV-2 cells regulates pro-inflammatory response and mitochondrial quality via mitophagy[J]. J Neuroimmunol, 2019, 336:577044.
[10] SHAO Q H, ZHANG X L, CHEN Y, et al. Anti-neuroinflammatory effects of 20C from Gastrodia elata via regulating autophagy in LPS-activated BV-2 cells through MAPKs and TLR4/Akt/mTOR signaling pathways[J]. Mol Immunol, 2018, 99:115-123.
[11] YAN J J, DU G H, QIN X M, et al. Baicalein attenuates the neuroinflammation in LPS-activated BV-2 microglial cells through suppression of pro-inflammatory cytokines, COX2/NF-κB expressions and regulation of metabolic abnormality[J]. Int Immunopharmacol, 2020, 79:106092.
[12] BAI Q, XUE M Z, YONG V W. Microglia and macrophage phenotypes in intracerebral haemorrhage injury:therapeutic opportunities[J]. Brain, 2020, 143(5):1297-1314.
[13] MA S, FAN L L, LI J C, et al. Resveratrol promoted the M2 polarization of microglia and reduced neuroinflammation after cerebral ischemia by inhibiting miR-155[J]. Int J Neurosci, 2020:1-9.
[14] BUTTURINI E, BORIERO D, CARCERERI DE PRATI A, et al. STAT1 drives M1 microglia activation and neuroinflammation under hypoxia[J]. Arch Biochem Biophys, 2019, 669:22-30.
[15] ZHU G W, CHENG Z B, LIN C L, et al. MyD88 regulates LPS-induced NF-kB/MAPK cytokines and promotes inflammation and malignancy in colorectal cancer cells[J]. Cancer Genomics Proteomics, 2019, 16(6):409-419.
[16] AKHTER N, HASAN A, SHENOUDA S, et al. TLR4/MyD88-mediated CCL2 production by lipopolysaccharide (endotoxin):Implications for metabolic inflammation[J]. J Diabetes Metab Disord, 2018, 17(1):77-84.
[17] KIM Y C, LEE S E, KIM S K, et al. Toll-like receptor mediated inflammation requires FASN-dependent MYD88 palmitoylation[J]. Nat Chem Biol, 2019, 15(9):907-916.
[18] LIU J T, WU S X, ZHANG H, et al. Inhibition of MyD88 signaling skews microglia/macrophage polarization and attenuates neuronal apoptosis in the hippocampus after status epilepticus in mice[J]. Neurotherapeutics, 2018, 15(4):1093-1111.
[19] KOBASHI S, TERASHIMA T, KATAGI M, et al. Transplantation of M2-deviated microglia promotes recovery of motor function after spinal cord injury in mice[J]. Mol Ther, 2020, 28(1):254-265.
[20] MI Y, WU Q, YUAN W R, et al. Role of microglia M1/M2 polarisation in the paraventricular nucleus:New insight into the development of stress-induced hypertension in rats[J]. Auton Neurosci, 2018, 213:71-80.
[21] WANG J, XING H Y, WAN L, et al. Treatment targets for M2 microglia polarization in ischemic stroke[J]. Biomed Pharmacother, 2018, 105:518-525.

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

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