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目的:观察缺氧对少突胶质前体细胞(preOL)内Nod样受体热蛋白结构域相关蛋白3(NLRP3)及细胞凋亡相关斑点样蛋白(ASC)表达的影响。方法:将体外分离纯化的preOL分为正常组和缺氧组,1%O25%CO2、37℃缺氧培养箱培养9 h建立preOL缺氧损伤模型。TUNEL法检测细胞凋亡,免疫荧光染色、qRT-PCR及Western Blot检测NLRP3表达,Western Blot检测ASC表达。结果:preOL缺氧损伤后胞质内出现空泡,胞膜破裂,突起出现肿胀、断裂;凋亡率较正常组增加(P<0.05);缺氧组NLRP3阳性细胞数和平均荧光强度均较正常组增加(P<0.05),NLRP3的mRNA及蛋白水平也均较正常组增加(P<0.05);ASC蛋白的表达在缺氧后上调(P<0.05)。结论:preOL的缺氧损伤可能与激活NLRP3炎症小体有关。
Objective: To observe the effect of hypoxia on the expression of NLRP3 and ASC in pre-oligodendrocyte precursor cells (preOL). Methods: The preOL isolated and purified in vitro was divided into normal group and hypoxia group, cultured in hypoxia incubator at 37 ℃ with 1% O25% CO2 for 9 h to establish hypoxic preox model. TUNEL method was used to detect apoptosis. Immunofluorescence staining, qRT-PCR and Western Blot were used to detect the expression of NLRP3. The expression of ASC was detected by Western Blot. Results: After hypoxia preoll injury, vacuoles and membrane rupture appeared in the cytoplasm, swelling and rupture of the protuberances were observed. The apoptosis rate of preOL hypoxia group was higher than that of the normal group (P <0.05). The number of NLRP3 positive cells and the average fluorescence intensity The mRNA and protein levels of NLRP3 were also increased in the normal group (P <0.05), compared with the normal group (P <0.05). The expression of ASC protein was up-regulated after hypoxia (P <0.05). Conclusion: The hypoxic injury of preOL may be related to the activation of NLRP3 inflammasome.