摘要
小胶质细胞的激活可导致神经元损伤,从而导致神经退行性疾病如帕金森病的发病。为了探讨毒死蜱(CPF)对小胶质细胞激活、炎症因子表达分泌,以及对多巴胺能神经元存活率的影响,本研究采用逆转录-聚合酶链反应(RT-PCR)、Western blot检测CPF刺激BV-2细胞(0、1、3、6、12、24h)后BV-2细胞形态的改变及诱导型一氧化氮合成酶(iNOS)、环氧化酶2(COX-2)mRNA和蛋白表达的变化;然后将BV-2细胞分为CPF组、DMSO对照组、NS对照组,分别刺激BV-2细胞12h后,将各BV-2细胞培养液加入SH-SY5Y细胞共培养24h。采用MTT方法检测经不同处理的BV-2细胞的条件培养液对SH-SY5Y存活率的影响。发现CPF作用下,BV-2小胶质细胞激活,炎症因子iNOS mRNA、COX-2mRNA及其蛋白表达水平均升高。激活的BV-2细胞通过分泌炎症因子造成神经元损伤。总之,CPF作用下小胶质细胞激活产生炎症因子,介导对神经元的损伤,进而可能参与了帕金森病的发病过程。
Excessive microglial activation and subsequent neuroinflammation lead to neuronal cell death, which are involved in the pathogenesis and progression of several neurodegenerative diseases such as Parkinson＇ disease. The objective of this study was to determine the involvement of chlorpyrifos （CPF） in the activation of microglia and production of inflammatory factors in response to CPF stimulation and the influence on the viability of dopaminergic （DA） neurons. We detected the change of BV-2 cells morphology and expression of inducible nitric oxide （iNOS）, cyclooxygenase-2 （COX-2） mRNA and protein level upon CPF stimulation （0, 1, 3, 6, 12, 24 h） in BV-2 （mouse brain microglia） cells by reverse transcription polymerase chain reaction （RT-PCR） or Western blot. We randomly assigned BV-2 cells into CPF, menstruum dimethysulfoxide （DMSO） and normal saline （NS） groups. We stimulated The BV-2 cells in the CPF group with CPF, and we stimulated the two control groups with DMSO or NS for 12 hours, respectively. We then collected the used culture media from the culture dishes and centrifuged it to remove the detached cells. Then, we used the supernatants as microglial conditioned media. We treated SH-SY5Y neurons with various groups of microglial conditioned media for 24 hours. We observed the effect of conditioned media collected from BV-2 cell on the viability of dopaminergic cell lines SH-SYSY using MTT assay. We found that inflammatory factors iNOS, COX-2 mRNA and protein levels were up-regulated upon CPF stimulation. Conditioned media from BV-2 upon CPF stimulation is toxic to SH-SYSY. It might be concluded that the exposure to CPF may induce dopaminergic neuronal damage by the activation of inflammatory response, and a mechanism may be involved in Parkinson＇s disease pathogenesis.
出处
《生物医学工程学杂志》
EI
CAS
CSCD
北大核心
2016年第3期506-511,共6页
Journal of Biomedical Engineering
