NLRP3炎症小体通路在IL-6、TNF-α促进椎间盘髓核细胞神经营养因子和感觉神经肽P物质表达中的作用

The role of NLRP3 inflammatome pathway in the expression of neurotrophic factor and substance P in⁃duced by IL-6 and TNF-αin nucleus pulposus cells from intervertebral disc

目的探讨NLRP3炎症小体通路在细胞炎症因子IL-6和TNF-α促进椎间盘髓核细胞(NP细胞)神经营养因子(NGF)和感觉神经肽P物质(SP)表达中的作用。方法从3只Sprague-Dawley大鼠椎间盘组织中提取原代NP细胞,分为对照组、处理组和MCC950组。对照组不经过任何处理,处理组用浓度1、10和100 ng/mL的IL-6或TNF-α处理48 h,MCC950组用1μmol/L的NLRP3抑制剂MCC950处理1 h后,用100 ng/mL的IL-6或TNF-α处理48 h。采用ELISA检测细胞培养上清液中NGF和SP的水平。结果与对照组相比,1、10和100 ng/mL IL-6或TNF-α处理NP细胞后,NGF和SP蛋白水平升高(P均<0.05)。MCC950组NGF、SP蛋白水平均较100 ng/mL IL-6或TNF-α处理组降低(P均<0.05)。结论细胞炎性因子IL-6和TNF-α能够通过激活NP细胞中NLRP3炎症小体信号通路,促进NGF和SP的表达。

Objective In order to study the role of NLRP3 inflammatome pathway in the expression of neurotrophic factor(NGF)and substance P(SP)induced by IL-6 and TNF-αin nucleus pulposus cells from intervertebral disc.Meth⁃ods Primary NP cells were extracted from the intervertebral disc tissues of three Sprague-Dawley rats.NP cells were di⁃vided into the control group,experimental group,and MCC950 group.The control group was non-treated.The experimen⁃tal groups of cells were treated with 1,10,and 100 ng/mL IL-6 or TNF-αfor 48 h.MCC950 group of cells were treated with 1μmol/L MCC950,an inhibitor of NLRP3,for 1 h and then treated with 100 ng/mL IL-6 or TNF-αfor 48 h.ELISA was performed for detecting the levels of NGF and SP in cell culture supernatant.Results Compared to the control group,1,10 and 100 ng/mL IL-6 or TNF-αtreatment in NP cells significantly increased the levels of NGF and SP(all P<0.05).Compared with 100 ng/mL IL-6 or TNF-αexperimental groups,NGF and SP levels in MCC950 group were lower(all P<0.05).Conclusion Cytokines IL-6 and TNF-αcan promote the expression of NGF and SP by activating the NL⁃RP3 inflammasome signaling pathway in NP cells.