灯盏乙素通过STAT3信号对激活的BV-2小胶质细胞M2型极化的影响

doi:10.13418/j.issn.1001-165x.2023.1.08

摘要/Abstract

摘要： 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。

关键词: 灯盏乙素； , , BV-2小胶质细胞； , , STAT3； , , 极化

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.

Key words: Scutellarin; , , BV-2 microglia; , , STAT3; , , Polarization

中图分类号:

R363.21

贾秋叶, 张晓丽娜, 陈浩伦, 袁云, 吴春云. 灯盏乙素通过STAT3信号对激活的BV-2小胶质细胞M2型极化的影响[J]. 中国临床解剖学杂志, 2023, 41(1): 36-44.

Jia Qiuye, Zhang Xiaolina, Chen Haolun, Yuan Yun, Wu Chunyun. Effect of Scutellarin on M2-type polarization in activated BV-2 microglia via STAT3 signaling[J]. Chinese Journal of Clinical Anatomy, 2023, 41(1): 36-44.

参考文献 20

[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.