孙静, 刘漪沦, 李灿, 等.hADMSCs影响TLR4-TRIF信号通路诱导小鼠小胶质细胞表型极化的实验研究.四川大学学报(医学版),2019,50(2):164-170
hADMSCs影响TLR4-TRIF信号通路诱导小鼠小胶质细胞表型极化的实验研究
Effect of Human Adipose Mesenchymal Stem Cells on Phenotype Polarization of Mice Microglia via TLR3/TRIF Signal Pathway
  
中文关键词:  人脂肪间充质干细胞表型Toll样受体
英文关键词:Human adipose derived mesenchymal stem cells (hADMSCs)PhenotypeToll-like receptors (TLRs)
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中文摘要:
      目的研究人脂肪间充质干细胞(human adipose-derived mesenchymal stem cells,hADMSCs)对小胶质细胞表型极化的影响及机制。方法取 C57/BL6小鼠BV-2小胶质细胞,以 hADMSCs+脂多糖(LPS)间接共培养,或以单纯LPS 培养。倒置显微镜下观察细胞形态。CCK-8法检测小胶质细胞增殖能力,实时荧光定量PCR(RT-qPCR)检测小胶质细胞M1/M2表型标记物的影响,Western blot检测小胶质细胞Toll样受体4(TLR4)-β干扰素TIR结构域衔接蛋白(TRIF)信号通路相关蛋白的表达。结果与单纯LPS培养相比,hADMSCs 加入后,小胶质细胞镜下形态相似, 细胞增殖活性被抑制(P<0.05),M1表型标记物的基因表达减少(P<0.05), M2表型标记物的基因表达增加(P<0.05),TRIF、TLR4、干扰素调节因子3 (IRF3)和磷酸化IRF3 (P-IRF3)蛋白的表达水平降低(P<0.05)。结论hADMSCs可抑制脂多糖(LPS)诱导的BV2小胶质细胞M1促炎表型的极化,从而诱导其向保护型M2表型极化,此作用可能与其对TLR4-TRIF信号通路活化的抑制有关。
英文摘要:
      ObjectiveTo explore the effect and mechanism of human adipose-derived mesenchymal stem cells (hADMSCs) on phenotypic polarization of microglia. MethodsBV-2 microglia of C57/BL6 mice were co-cultured with hADMSCs+lipopolysaccharide (LPS), or cultured with LPS alone. Cell morphology was observed under an inverted microscope. The effect of hADMSCs on microglial proliferation was evaluated by CCK-8 assay. The impact of hADMSCs on microglia M1/M2 phenotype markers were detected using quantitative real-time PCR (RT-qPCR). The affect of hADMSCs on the proteins expression levels of Toll-like receptor 4 (TLR4)-TIR domain containing adaptor protein inducing interferon β (TRIF) signaling pathway in BV-2 microglia was detected by using Western blot analysis. ResultsAs compared with the LPS treatment, hADMSCs treatment had no obvious effect on microglia morphology, whereas showed significant inhibition on microglial proliferation activity (P<0.05). Simultaneously, hADMSCs treatment reduced expression of microglia M1 phenotype markers (P<0.05), and increased microglia M1 phenotype markers in gene levels (P<0.05). At the same time, protein expression levels of TRIF, TLR4, phosphorylated interferon regulatory factor 3 (P-IRF3) and interferon regulatory factor 3 (IRF3) in BV-2 microglia were decreased after hADMSCs treatment. ConclusionhADMSCs can blockade the LPS-induced pro-inflammatory microglia M1 phenotype, whereas induces protective microglial M2 phenotype, which may be related to inhibition of the TLR4-TRIF signaling pathway.
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