纳米二氧化钛对小胶质细胞Notch信号通路及炎症因子分泌水平的影响

Effects of Nano-titanium Dioxide on Notch Signaling Pathway and Secretion of Inflammatory Factors in Microglia

由于纳米材料的广泛应用及其可能存在的生物安全性风险,本研究探讨了纳米二氧化钛对小胶质细胞Notch信号通路及炎症因子分泌水平的影响。以不同浓度的纳米二氧化钛染毒小胶质细胞,MTT法测定细胞活力,乳酸脱氢酶(LDH)检测试剂盒测定细胞培养液上清液LDH活性,ELISA法测定细胞培养液上清液肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)的分泌水平,Western Blot法检测Notch-1和Hes-1的蛋白表达水平。结果表明,与对照组相比,纳米二氧化钛40.0μg·mL^(-1)和50.0μg·mL^(-1)暴露组细胞活力显著降低;纳米二氧化钛20.0、30.0和40.0μg·mL^(-1)暴露组LDH水平明显升高;纳米二氧化钛15.0、20.0、30.0和40.0μg·mL^(-1)暴露组TNF-α、IL-1β和IL-6分泌水平升高;纳米二氧化钛20.0、30.0和40.0μg·mL^(-1)暴露组Notch-1及Hes-1蛋白表达水平升高。研究表明,纳米二氧化钛暴露导致细胞活力降低,破坏细胞膜的完整性,炎症因子及Notch信号通路相关蛋白Notch-1和Hes-1的表达水平升高。

In consideration of the wide application of nanomaterials and the potential risk of their biosafety,this study aimed to investigate the effects of Nano-TiO_(2)on Notch signaling pathway and the secretion of inflammatory factors in microglia.Different concentrations of Nano-TiO_(2)were used to intervene microglia cells.The cell viability was determined by MTT;the lactate dehydrogenase(LDH)detection kit was used to determine LDH activity in the cell supernatant;the secretion level of tumor necrosis factor-α(TNF-α),interleukin-1β(IL^(-1)β)and interleukin-6(IL-6)in the cell supernatant were measured by ELISA;the protein expression levels of Notch-1 and Hes-1 were detected by Western Blotting.Compared with the control group,the cell viability of the 40.0μg·mL^(-1)and 50.0μg·mL^(-1)Nano-TiO_(2)exposure group was significantly reduced;20.0,30.0 and 40.0μg·mL^(-1)Nano-TiO_(2)exposure group had a significant increase in LDH level;the secretion levels of TNF-α,IL^(-1)βand IL-6 were increased in the 15.0,20.0,30.0 and 40.0μg·mL^(-1)Nano-TiO_(2)exposure groups;the expression levels of Notch-1 and Hes-1 were increased in the 20.0,30.0 and 40.0μg·mL^(-1)Nano-TiO_(2)exposure groups.It is suggested that the exposure of Nano-TiO_(2)lead to the decrease of cell viability,the destruction of cell membrane integrity,and the increase on the expression of inflammatory factors,Notch-1 and Hes-1 proteins.