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| 右美托咪定通过c-Fos/NLRP3/caspase-1级联抑制LPS诱发的小胶质细胞炎症反应 |
| Dexmedetomidine inhibits LPS-induced microglial inflammatory response via c-Fos/NLRP3/caspase-1 cascade |
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| DOI：10.12089/jca.2019.08.017 |
| 中文关键词: 右美托咪定 脂多糖 c-Fos NLRP3蛋白 caspase-1 炎症反应 小胶质细胞 |
| 英文关键词: Dexmedetomidine Lipopolysaccharide c-Fos NLRP3 protein Caspase-1 Inflammatory response Microglia |
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| 中文摘要: |
目的 探讨右美托咪定通过c-Fos/NLRP3/caspase-1级联抑制脂多糖（LPS）诱发的小胶质细胞炎症反应。
方法 选取新生的SD大鼠，取其小胶质细胞进行原代培养及分离纯化。采用LPS诱导建立小胶质细胞炎症模型。采用MTT法选取右美托咪定抑制LPS诱导小胶质细胞炎症反应的最适宜浓度。将细胞分为三组：对照组、LPS组和右美托咪定治疗组（D组）。采用实时定量PCR法测定细胞中IL-1β和TNF-α的mRNA表达量；采用ELISA法检测细胞上清液中IL-1β和TNF-α的含量；采用Western blot法检测原癌基因 c-Fos、NLRP3和caspase-1的蛋白含量。
结果 与对照组比较，LPS组小胶质细胞中炎性因子IL-1β和TNF-α的mRNA表达量均明显升高（P<0.05）；D组IL-1β和TNF-α的mRNA表达量明显低于LPS组（P<0.05）。与对照组比较，LPS组中小胶质细胞中炎性因子IL-1β和TNF-α的含量均明显增加（P<0.05）；而D组IL-1β和TNF-α的含量明显低于LPS组（P<0.05）。与对照组比较，LPS组小胶质细胞中c-Fos、NLRP3和caspase-1的蛋白含量均明显增加（P<0.01）；而D组c-Fos、NLRP3和caspase-1的蛋白含量明显低于LPS组（P<0.05）。
结论 右美托咪定可抑制LPS诱发的小胶质细胞中炎性因子IL-1β和TNF-α的mRNA表达量及蛋白含量，推测其作用机制可能与抑制c-Fos/NLRP3/caspase-1级联反应有关。 |
| 英文摘要: |
Objective To investigate the inhibition of LPS-induced microglial inflammatory response by dexmedetomidine (DEX) via c-Fos/NLRP3/caspase-1 cascade.
Methods Newborn SD rats were selected and their microglia was cultured and purified. Microglia was induced by 5 μg/ml LPS to establish a microglial inflammation model. The optimal concentration of DEX to inhibit LPS-induced microglial inflammatory response was determined by MTT method. The cells were divided into three groups: control group, LPS group and DEX group. Real-time quantitative PCR was used to determine the mRNA expression levels of inflammatory factors interleukin-1β (IL-1β) and tumor necrosis factor alpha (TNF-α) in cells; cell supernatant was detected by ELISA. The secretion of IL-1β and TNF-α in the liquid; the proto-oncogene c-Fos and nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) and caspase-1 protein expression levels were detected by Western blot.
Results Real-time quantitative PCR showed that compared with the control group, the inflammatory factors IL-1β and TNF-α in the microglia of the LPS group were significantly higher (P < 0.05). The mRNA expression levels of inflammatory factors IL-1β and TNF-α in the DEX group and cells were significantly lower than those in the LPS group (P < 0.05). The results of ELISA showed that compared with the control group, the secretion of IL-1β and TNF-α in LPS group were significantly incresed (P < 0.05). Compared with that of LPS group, the secretion of IL-1β and TNF-α was significantly lower (P < 0.05). The results of Western Blot showed that compared with the control group, the protein expression levels of c-Fos, NLRP3 and caspase-1 in glial cells in the LPS group were significantly increased (P < 0.01). Compared with those in LPS group, the protein expression levels of c-Fos, NLRP3 and caspase-1 in cells were significantly lower than those in the DEX group (P < 0.05).
Conclusion Dexmedetomidine can inhibit the mRNA expression and protein secretion of inflammatory cytokines IL-1β and TNF-α in LPS-induced microglia. The mechanism may be related to the inhibition of c-Fos/NLRP3/caspase-1 cascade. |
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