【摘要】 目的探讨IL-1β辅助小胶质细胞M1型活化的作用和机制。方法将BV2细胞分为对照（Control）组、脂多糖（LPS）组、LPS+IL-1β组。Control组常规培养;LPS组用终浓度为1 mg/L的LPS处理;LPS+IL-1β组先用15μg/L的IL-1β预处理6 h,再用终浓度为1 mg/L的LPS处理。LPS的处理时间为24 h。酪氨酸激酶（JAK1）抑制剂IN-7处理的实验中,将BV2细胞分为LPS组、LPS+IL-1β组、LPS+IL-1β+IN-7组,其中LPS+IL-1β+IN-7组在IL-1β处理前6 h用0.5μmol/L IN-7预处理,阻断JAK1,其余处理同前。采用ELISA法检测培养基中IL-6、TNF-α、一氧化氮（NO）和前列腺素G₂（PEG₂）水平;免疫荧光染色观察BV2细胞中CD86的荧光强度;Western blot法检测M1型标志物单核细胞趋化蛋白1（MCP-1）、趋化因子8（CXCL-8）的表达以及JAK1-信号转导及转录激活蛋白1（STAT1）信号蛋白JAK1、STAT1、磷酸化的JAK1（p-JAK1）、磷酸化的STAT1（p-STAT1）的表达。结果与LPS组比较,LPS+IL-1β组培养基中在3、6、12、18、24 h时IL-6、TNF-α、NO和PEG₂表达水平均上调（均P<0.05）,细胞中CD86荧光强度上调（P<0.05）,细胞中MCP-1、CXCL-8、JAK1、p-JAK1、p-STAT1蛋白表达水平均上调（均P<0.05）。而IN-7处理后,与LPS+IL-1β组比较,LPS+IL-1β+IN-7组培养基在3、6、12、18、24 h时IL-6、TNF-α、NO和PEG₂表达水平均下调（均P<0.05）,细胞中CD86荧光强度下调（P<0.05）,细胞中MCP-1、CXCL-8、JAK1、STAT1、p-JAK1、p-STAT1蛋白表达水平均下调（均P<0.05）。结论 IL-1β可以通过激活JAK1-STAT1信号,进一步促进小胶质细胞M1型活化,这是神经炎症进展的机制之一。更多还原

【Abstract】 Objective To study the effect and mechanism of interleukin-1β（IL-1β） on M1 polarization of microglia.Methods Mouse microglia BV2 cells were treated with lipopolysaccharide（LPS） for 24 h to induce the M1 polarization（LPS group）, or 6 h before LPS treatment BV2 cells were pretreated with IL-1β（LPS+IL-1β group）. The contents of IL-6, TNF-α, NO and PEG₂ were detected by enzyme-linked immunosorbent assay（ELISA）, CD86 in BV2 cells was observed by immunofluorescence, Western blot was used to detect the expression of MCP-1, CXCL-8, JAK1, STAT1, p-JAK1, and p-STAT1. After pretreatment of BV2 cells with JAK1 inhibitor IN-7, M1 polarization was induced with IL-1β and LPS. Cytokine expression, CD86 expression, and expression of marker proteins in the culture medium were examined. Results Compared with the LPS group,the expression levels of IL-6, TNF-α, NO and PEG₂ in the LPS+IL-1β group were up-regulated at 3, 6, 12, 18, and 24 h（all P<0.05）. The expression level of CD86 in cells was up-regulated（P<0.05）, and the protein expression levels of MCP-1, CXCL-8,JAK1, p-JAK1, and p-STAT1 in cells were up-regulated（all P<0.05）. After IN-7 treatment, compared with the LPS+IL-1β group,the expression levels of IL-6, TNF-α, NO and PEG₂ in the LPS+IL-1β+IN-7 group at 3, 6, 12, 18, and 24 h were down-regulated（all P <0.05）. The expression level of CD86 in cells was down-regulated（P <0.05）, and the protein expression levels of MCP-1, CXCL-8, JAK1, STAT1, p-JAK1, and p-STAT1 in cells were down-regulated（all P<0.05）. Conclusion IL-1β can further promote the M1 polarization of mouse microglia by activating JAK1-STAT1 signaling, which may be one of the mechanisms of neuroinflammation progression.更多还原