目的 探究灯盏乙素（Scutellarin）对脂多糖（Lipopolysaccharide，LPS）作用下M2型小胶质细胞极化的作用及信号转导和转录激活因子3（signal transducers and activators of transcription 3，STAT3）的调控机制。 方法 BV-2小胶质细胞分为对照组（Con组）、STAT3抑制剂α-氰基-（3，4-二羟基）-N-苄基霉素-烟酰胺组（AG490组）、LPS组、LPS+AG490组、LPS+灯盏乙素预处理组（LPS+S组）、LPS+S+AG490组，共6组。Western blot和免疫荧光染色检测STAT3、磷酸化STAT3（P-STAT3）和M2型小胶质细胞标记物白介素10（IL-10）的表达。 结果 Western blot结果显示，LPS组的P-STAT3、IL-10表达显著增强，与对照组差异显著（P<0.05）；使用灯盏乙素干预后P-STAT3表达明显降低；IL-10表达显著增高；均与LPS组有显著差异（P<0.05）。AG490增强灯盏乙素的作用。此外，STAT3在各组的变化无统计学意义（P>0.05）。免疫荧光染色结果显示，灯盏乙素干预后可降低LPS激活的BV-2小胶质细胞P-STAT3的蛋白表达水平；增强IL-10蛋白表达。各组STAT3的蛋白水平无统计学意义。 结论 灯盏乙素通过抑制STAT3活化促进BV-2小胶质细胞向M2型极化，可能与灯盏乙素减轻神经炎症反应有关。
Abstract
Objective To investigate the role of scutellarin on the polarization of M2 microglia under the action of lipopolysaccharide (LPS) and the regulation mechanism of signal transducers and activators of transcription 3 (STAT3). Methods BV-2 microglia were divided into a control group (Con), a STAT3 inhibitor α-cyano-(3,4-dihydroxy)-N-benzylmycin-nicotinamide group (AG490), a LPS group (LPS), a LPS+AG490 group, a LPS+scutellarin pretreatment group (LPS+S), a LPS+S+AG490 group. The expression of STAT3, phosphorylated STAT3 (P-STAT3) and the M2 microglia marker interleukin 10 (IL-10) were detected by Western Blot and immunofluorescence staining. Results Western Blot results showed: the expression of P-STAT3 and IL-10 in the LPS group was significantly enhanced, compared with that in the control group (P<0.05); the expression of P-STAT3 was significantly reduced, the expression of IL-10 was obviously increased after pretreatment with scutellarin; both were significantly different from the LPS group (P<0.05). AG490 enhanced the effect of scutellarin. In addition, there was no statistical difference s in STAT3 in each group (P>0.05). Immunofluorescence staining results showed that scutellarin intervention reduced protein level of P-STAT3 in LPS-activated BV-2 microglia, enhanced protein level of IL-10. There was no statistical difference in protein level of STAT3 in each group. Conclusions Scutellarin promotes the polarization of BV-2 microglia to M2 type by inhibiting the activation of STAT3, which may be related to the effect of scutellarin in reducing neuro-inflammatory responses.
关键词
灯盏乙素； /
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BV-2小胶质细胞； /
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STAT3； /
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极化
Key words
Scutellarin; /
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BV-2 microglia; /
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STAT3; /
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Polarization
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参考文献
[1] Price CJ, Wang D, Menon DK, et al. Intrinsic activated microglia map to the peri-infarct zone in the subacute phase of ischemic stroke[J]. Stroke, 2006, 37(7): 1749-53. DOI: 10.1161/01.STR.0000226980.95389.0b.
[2] Lang GP, Li C, Han YY. Rutin pretreatment promotes microglial M1 to M2 phenotype polarization[J]. Neural Regen Res, 2021, 16(12): 2499-2504. DOI: 10.4103/1673-5374.313050.
[3] Ham HJ, Han SB, Yun J, et al. Bee venom phospholipase A2 ameliorates amyloidogenesis and neuroinflammation through inhibition of signal transducer and activator of transcription-3 pathway in Tg2576 mice[J]. Transl Neurodegener, 2019, 8: 26. DOI: 10.1186/s40035-019-0167-7.
[4] Zhou S, Guo X, Chen S, et al. Apelin-13 regulates LPS-induced N9 microglia polarization involving STAT3 signaling pathway[J]. Neuropeptides, 2019, 76: 101938. DOI: 10.1016/j.npep.2019.101938.
[5] Yu CI, Cheng CI, Kang YF, et al. Hispidulin inhibits neuroinflammation in lipopolysaccharide-activated BV2 microglia and attenuates the activation of Akt, NF-κB, and STAT3 pathway[J]. Neurotox Res, 2020, 38(1): 163-174. DOI: 10.1007/s12640-020-00197-x.
[6] Pengyue Z, Tao G, Hongyun H, et al. Breviscapine confers a neuroprotective efficacy against transient focal cerebral ischemia by attenuating neuronal and astrocytic autophagy in the penumbra[J]. Biomed Pharmacother, 2017, 90: 69-76. DOI: 10.1016/j.biopha. 2017. 03.039.
[7] Chen HL, Jia WJ, Li HE, et al. Scutellarin exerts anti-inflammatory effects in activated microglia/brain macrophage in cerebral ischemia and in activated BV-2 microglia through regulation of mapks signaling pathway[J]. Neuromolecular Med, 2020, 22(2): 264-277. DOI: 10.1007/s12017-019-08582-2.
[8] Zhao Y, Gan Y, Xu G, et al. Exosomes from MSCs overexpressing microRNA-223-3p attenuate cerebral ischemia through inhibiting microglial M1 polarization mediated inflammation[J]. Life Sci, 2020, 260: 118403. DOI: 10.1016/j.lfs.2020.118403.
[9] Weng L, Zhang H, Li X, et al. Ampelopsin attenuates lipopolysaccharide-induced inflammatory response through the inhibition of the NF-κB and JAK2/STAT3 signaling pathways in microglia[J]. Int Immunopharmacol, 2017, 44: 1-8. DOI: 10.1016/j.intimp.2016.12.018.
[10]Yuan Y, Zha H, Rangarajan P, et al. Anti-inflammatory effects of Edaravone and Scutellarin in activated microglia in experimentally induced ischemia injury in rats and in BV-2 microglia[J]. BMC Neurosci, 2014, 15: 125. DOI: 10.1186/s12868-014-0125-3.
[11]Xu H, Wang E, Chen F, et al. Neuroprotective phytochemicals in experimental ischemic stroke: mechanisms and potential clinical applications[J]. Oxid Med Cell Longev, 2021, 2021: 6687386. DOI: 10.1155/2021/6687386.
[12] Schäbitz WR, Kollmar R, Schwaninger M, et al. Neuroprotective effect of granulocyte colony-stimulating factor after focal cerebral ischemia[J]. Stroke, 2003, 34(3): 745-751. DOI: 10.1161/01.STR. 0000057814. 70180.17.
[13] Kong LN, Lin X, Huang C, et al. Hesperetin derivative-12 (HDND-12) regulates macrophage polarization by modulating JAK2/STAT3 signaling pathway[J]. Chin J Nat Med, 2019, 17(2): 122-130. DOI: 10.1016/S1875-5364(19)30014-7.
[14] Zhou HJ, Yang X, Cui HJ, et al. Leukemia inhibitory factor contributes to reactive astrogliosis via activation of signal transducer and activator of transcription 3 signaling after intracerebral hemorrhage in rats[J]. J Neurotrauma, 2017, 34(8): 1658-1665. DOI: 10.1089/neu.2016.4711.
[15] Fang YY, Zhang JH. MFG-E8 alleviates oxygen-glucose deprivation-induced neuronal cell apoptosis by STAT3 regulating the selective polarization of microglia[J]. Int J Neurosci, 2021, 131(1): 15-24. DOI: 10.1080/00207454.2020.1732971.
[16] Wu J, Sun L, Li H, et al. Roles of programmed death protein 1/programmed death-ligand 1 in secondary brain injury after intracerebral hemorrhage in rats: selective modulation of microglia polarization to anti-inflammatory phenotype[J]. J Neuroinflammation, 2017, 14(1): 36. DOI: 10.1186/s12974-017-0790-0.
[17] Bode JG, Ehlting C, Häussinger D. The macrophage response towards LPS and its control through the p38(MAPK)-STAT3 axis[J]. Cell Signal, 2012, 24(6): 1185-1194. DOI: 10.1016/j.cellsig.2012.01.018.
[18] Guo H, Fan Z, Wang S, et al. Astrocytic A1/A2 paradigm participates in glycogen mobilization mediated neuroprotection on reperfusion injury after ischemic stroke[J]. J Neuroinflammation, 2021, 18(1): 230. DOI: 10.1186/s12974-021-02284-y.
[19] Lisi L, Stigliano E, Lauriola L, et al. Proinflammatory-activated glioma cells induce a switch in microglial polarization and activation status, from a predominant M2b phenotype to a mixture of M1 and M2a/B polarized cells[J]. ASN Neuro, 2014, 6(3): 171-183. DOI: 10.1042/AN20130045.
[20] Mishra SK, Kumar BS, Khushu S, et al. Early monitoring and quantitative evaluation of macrophage infiltration after experimental traumatic brain injury: a magnetic resonance imaging and flow cytometric analysis[J]. Mol Cell Neurosci, 2017, 78: 25-34. DOI: 10.1016/j.mcn.2016.11.008.
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