N-乙酰-半胱氨酸对炎症反应中BV-2细胞的保护作用

Protective effect of N-acetyl-cysteine on BV-2 cells in inflammatory response

为了明确N-乙酰-半胱氨酸(NAC)在LPS诱导的BV-2细胞炎症反应中的保护作用,本研究利用流式细胞术检测细胞活性氧(ROS)、超氧阴离子自由基(O2·-)和线粒体超氧化物(MitoSOX)水平;ELISA和实时荧光定量PCR法检测IL-1β、IL-6和TNF-α表达;脂质氧化产物丙二醛(MDA)试剂盒检测细胞MDA含量;流式细胞术检测细胞凋亡水平。通过对细胞氧化应激、促炎因子表达、脂质损伤和细胞凋亡的检测,探讨NAC对BV-2细胞炎症反应的保护作用。结果显示,LPS可以使BV-2细胞总ROS、O2·-和MitoSOX的表达显著上升,加入NAC后ROS、O2·-和MitoSOX的水平又显著下降。LPS可使BV-2细胞IL-1β、IL-6和TNF-α的表达量显著升高,加入NAC后IL-1β、IL-6和TNF-α的表达量显著下降(P<0.05)。LPS处理4 h后细胞MDA含量显著增加(P<0.05),细胞凋亡水平升高,而加入NAC后MDA含量及细胞凋亡水平显著下降。结果表明,NAC可以抑制LPS引起的BV-2细胞氧化应激,使促炎因子表达减少,同时使BV-2细胞脂质损伤减弱和细胞凋亡水平下降,NAC可对细胞炎症反应中BV-2细胞起到很好的保护作用。

To investigate the effect of N-acetyl-cysteine(NAC)on BV-2 cells in inflammatory response,flow cytometry(FCM)was used to detect the level of total ROS,O2·-and MitoSOX levels.ELISA or qRT-PCR were used to detect the protein secretion and mRNA expression of IL-1β,IL-6 and TNF-α.Lipid peroxidation MDA assay Kit was used to detect the level of MDA content.FCM was used to detect the level of apoptosis.By examining the effects of NAC on oxidative stress,inflammatory factor expression,lipid damage and apoptosis in BV-2 cells under inflammatory response,the effect of NAC on BV-2 cells under inflammatory response was investigated.The results showed that LPS could induce a significant increase in ROS,O2·-and MitoSOX in BV-2 cells,the levels of ROS,O2·-and MitoSOX were significantly decreased after the addition of NAC.MDA content increased after LPS treatment of BV-2 cells for 4 h(P<0.05),and the level of apoptosis increased.MDA content and apoptosis levels were significantly reduced after the addition of NAC.The results showed that NAC could inhibit the oxidative stress,inflammatory factor expression,lipid damage and apoptosis of BV-2 cells induced by LPS.It indicated that NAC had protective effect during the process of BV-2 cell inflammatory response.