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该文探讨β-细辛醚对Aβ1-42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RA-h/PC12共培养体系,实时动态观察Aβ1-42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Aβ1-42作用于RA-h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IL-1β,TNF-α,BDNF的水平;Western blot法检测PC12细胞NF-κB的活性和ERK,p38,JNK磷酸化水平。结果显示,β-细辛醚(55.5 mg·L-1)能显著减缓Aβ1-42诱导的RA-h活化引起的PC12细胞存活率下降(P<0.01),显著降低下室培养液中IL-1β,TNF-α的水平和PC12细胞ERK,p38,JNK磷酸化水平(P<0.01);β-细辛醚(166.7 mg·L-1)能促进下室培养液中BDNF的释放(P<0.05)。以上结果表明,Aβ1-42诱导RA-h活化并释放IL-1β,TNF-α等炎症因子加剧PC12细胞的损伤;β-细辛醚能降低IL-1β,TNF-α和促进BDNF的释放,抑制PC12细胞NF-κB的活性和ERK,p38,JNK磷酸化水平。
This article is to explore the protective effect of β-asarone on PC12 cell injury induced by Aβ1-42-induced astrocyte activation and its mechanism for the prevention and treatment of Alzheimer’s disease (AD) In the application of experimental basis. First, real-time marker-free dynamic cell analysis (RTCA) was used to construct RA-h / PC12 co-culture system. The survival of PC12 cells in co-culture system was observed in real time after Aβ 1-42 -induced astrocyte injury Rate, the optimal drug intervention time was selected according to the survival rate curve and parameters automatically generated by the system, astrocytes were intervened by different concentrations of β-asarone, and the survival rate of PC12 cells in co-culture system was observed . Secondly, the effect of Aβ1-42 on the survival rate of PC12 cells and the effect of β-asarone on the survival rate of PC12 cells were detected by MTT assay, and the detection results of RTCA were further verified by ELISA. IL-1β, TNF-α and BDNF were detected by Western blot. The phosphorylation of NF-κB and phosphorylation of ERK, p38, JNK in PC12 cells were detected by Western blot. The results showed that β-asarone (55.5 mg · L-1) significantly decreased the survival rate of PC12 cells induced by Aβ1-42-induced RA-h activation (P <0.01) 1β, TNF-α and phosphorylation levels of ERK, p38 and JNK in PC12 cells (P <0.01), while β-asarone (166.7 mg · L -1) promoted the release of BDNF in lower chamber culture medium (P < 0.05). The above results indicate that Aβ1-42 induces activation of RA-h and releases inflammatory factors such as IL-1β and TNF-α, which can aggravate the injury of PC12 cells. Β-asarone can decrease the release of IL-1β, TNF-α and promote the release of BDNF, Inhibit the activity of NF-κB and phosphorylation of ERK, p38, JNK in PC12 cells.