微小RNA-155对小胶质BV-2细胞炎症因子分泌及吲哚胺2,3-双加氧酶表达的影响  

Effects of miR-155 over-expression on expression of inflammatory factors and indolamine 2, 3-dioxygenase in microglial BV-2 cells

作　　者：孙晓花[1] 尹利明[2] 赵燕娜[2] 宋明芬[1] 宋海东[1] SUN Xiao-hua;YIN Li-ming;ZHAO Yan-na;SONG Ming-fen;SONG Hai-dong(Hangzhou Seventh People’s Hospital, Hangzhou 310013, China;The Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou 310006, China)

机构地区：[1]杭州市第七人民医院,浙江杭州310013 [2]浙江中医药大学附属第一医院,浙江杭州310006

出　　处：《中国病理生理杂志》2019年第8期1403-1408,共6页Chinese Journal of Pathophysiology

基　　金：浙江省自然科学基金资助项目(No.LQ16H090008)

摘　　要：目的:探讨微小RNA-155(miR-155)对小胶质BV-2细胞炎症因子分泌及吲哚胺2,3-双加氧酶(IDO)表达的影响。方法:采用携带miR-155的慢病毒感染小胶质BV-2细胞,以脂多糖(LPS)处理BV-2细胞为对照。观察细胞形态,流式液相芯片检测炎症因子的分泌水平,real-time PCR检测炎症因子和IDO的mRNA表达水平,Western blot法检测细胞因子信号抑制物1(SOCS1)、磷酸化p38 MAPK和IDO蛋白的蛋白水平。结果:携带miR-155的慢病毒成功感染BV-2细胞,其miR-155表达水平高于LPS处理组和阴性病毒感染组(P<0.01)。miR-155促进BV-2细胞白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)、单核细胞趋化蛋白1(MCP-1)和IL-10分泌,抑制IL-12分泌,上调IL-6、TNF-α、IL-10和IDO的mRNA表达,同时升高IDO蛋白表达水平和p38 MAPK蛋白磷酸化水平,下调SOCS1蛋白表达(P<0.01)。LPS刺激BV-2细胞分泌炎症因子IL-6、TNF-α、MCP-1和IL-12,上调IL-6、TNF-α和IDO mRNA表达,同时上调IDO、p-p38 MAPK和SOCS1的蛋白水平。结论:miR-155促进小胶质BV-2细胞相关炎症因子分泌和IDO蛋白表达,可能与SOCS1和p38 MAPK信号通路相关。AIM: To explore the effect of microRNA-155(miR-155)over-expression on the expression of inflammatory factors and indolamine 2, 3-dioxygenase(IDO) in the microglial BV-2 cells. METHODS: For over-expression of miR-155, the BV-2 cells were transfected with lentiviral vector carrying mmu-miR-155. The expression of inflammatory factors was detected by cytometric bead array system(CBA). The mRNA expression of inflammatory factors and IDO was analyzed by real-time PCR. The protein levels of suppressor of cytokine signalling 1(SOCS1), p-p38 MAPK and IDO were determined by Western blot. RESULTS:The expression of miR-155 was up-regulated in the BV-2 cells transfected with lentiviral vector carrying mmu-miR-155 compared with LPS treatment group(P<0.01). The miR-155 over-expression promoted the secretion of interleukin-6(IL-6), tumor necrosis factor-α(TNF-α), monocyte chemotactic protein-1(MCP-1) and IL-10, and inhibited the secretion of IL-12. The miR-155 over-expression increased the mRNA expression of IL-6, TNF-α, IL-10 and IDO, also increased the protein levels of IDO and p-p38 MAPK, but decreased the protein expression of SOCS1(P<0.01). LPS promoted the secretion of IL-6, TNF-α, MCP-1 and IL-12, also increased the mRNA expression of IL-6, TNF-α and IDO, meanwhile, increased the protein levels of IDO, p-p38 MAPK and SOCS1(P<0.01). CONCLUSION: Over-expression of miR-155 promotes the secretion of related imflammatory factors and protein expression of IDO in microglial BV-2 cells mediated with SOCS1 and p38 MAPK signaling pathway.

关 键 词：微小RNA-155 BV-2细胞 炎症因子 吲哚胺2 3-双加氧酶 P38 MAPK信号通路 

分 类 号：R329.25[医药卫生—人体解剖和组织胚胎学] R363.2[医药卫生—基础医学]