β蚕感撩讯Aβ142诱导星形胶质细胞活化所致PC12细胞损伤保护作用研究.docVIP

β蚕感撩讯Aβ142诱导星形胶质细胞活化所致PC12细胞损伤保护作用研究 　　[摘要]该文探讨β细辛醚对Aβ142诱导星形胶质细胞活化所致PC12细胞损伤的保护作用及其机制，为β细辛醚在阿尔兹海默病（Alzheimer’s diease，AD）防治中的应用提供实验依据。首先，采用实时无标记动态细胞分析技术（Real time cell analysis，RTCA）构建RAh/PC12共培养体系，实时动态观察Aβ142诱导星形胶质细胞损伤后对共培养体系中PC12细胞存活率的影响，根据系统自动生成的存活率曲线和参数选出最佳的药物干预时间，采用不同浓度的β细辛醚对星形胶质细胞进行干预，观察共培养体系中PC12细胞存活率的改变；其次，采用MTT法检测Aβ142作用于RAh对PC12细胞存活率的影响以及β细辛醚的干预效应，验证RTCA的检测结果；进一步采用ELISA法检测下室培养液中IL1β，TNFα，BDNF的水平；Western blot法检测PC12细胞NFκB的活性和ERK，p38，JNK磷酸化水平。结果显示，β细辛醚（555 mg?L-1）能显著减缓Aβ142诱导的RAh活化引起的PC12细胞存活率下降（P001），显著降低下室培养液中IL1β，TNFα的水平和PC12细胞ERK，p38，JNK磷酸化水平（P001）；β细辛醚（1667 mg?L-1）能促进下室培养液中BDNF的释放（P005）。以上结果表明，Aβ142诱导RAh活化并释放IL1β，TNFα等炎症因子加剧PC12细胞的损伤；β细辛醚能降低IL1β，TNFα和促进BDNF的释放，抑制PC12细胞NFκB的活性和ERK，p38，JNK磷酸化水平。 　　[关键词]β细辛醚；Aβ142；星形胶质细胞；PC12细胞；RTCA；NFκB 　　[Abstract]This study was aimed to investigate the protective effect and mechanism of βasarone on PC12 cells injury induced byAβ142 activated astrocytes， and provide experimental basis for βasarone application in the prevention and control of Alzheimer′s disease （AD） Firstly， RAh and PC12 cells were cocultured in the special transwell chamber， and the Real time cell analysis （RTCA） system was used to realtime observe its effect on PC12 cells survival rate in the coculture system after astrocytes injury induced by Aβ142. The best intervention time of βasarone was selected according to the survival curve and parameters generated automatically βasarone with different concentrations was used for intervention on astrocytes， then the changes of PC12 cells survival rate in the coculture system were observed Secondly， MTT assay was used to detect the effect of Aβ142 on PC12 cells survival rate as well as the intervention effect of βasarone， and verify the testing results of RTCA The levels of IL1β， TNFα and BDNF in culture media of the lower chamber were detected by ELISA The NFκB activity and phosphorylation levels of ERK， p38 and JNK were detected by Western blot Results showed that βasarone （555 mg?L-1） could significantl