多奈哌齐对PC12细胞淀粉样前体蛋白α代谢途径的影构

发布时间：2018-10-30 12:33

【摘要】：目的观察多奈哌齐对PC12细胞淀粉样前体蛋白（APP）非淀粉样物质代谢途径（α途径）的影响以及可能的细胞信号机制。方法以体外培养的PC12细胞为观察对象,分为空白对照组、多奈哌齐（20μmol/L）组、多奈哌齐（20μmol/L）+PI3-K抑制剂LY294002组(LY组)、多奈哌齐（20μmol/L）+P38抑制剂SB203580组(SB组)、多奈哌齐（20μmol/L）+JNK抑制剂SP600125组(SP组)。各抑制剂组在加入多奈哌齐前30分钟分别给予相应的抑制剂。培养24h后，用Western blot检测细胞培养上清液中的APP、ADAM10和ADAM17蛋白水平；ELISA方法检测细胞培养上清液中的sAPPα和sAPP/β水平。结果 1. APP蛋白表达：空白对照组、多奈哌齐组、LY组、SB组和SP组的APP蛋白表达水平，各组间比较无统计学意义（P＞0.05）。 2. ADAM10和ADAM17蛋白：用Western blot检测ADAM10和ADAM17蛋白。与对照组相比较,多奈哌齐组、LY组、SB组和SP各组ADAM10和ADAM17均有增加，差异均有统计学意义（P＜0.05）。各抑制剂组与多奈哌齐组比较，LY组ADAM10和ADAM17表达无明显差异（P＞0.05），SB及SP组ADAM10和ADAM17表达减少，差异有统计学意义（P＜0.05）。 3. sAPPα和sAPPβ表达：用ELISA检测sAPPα及sAPPβ蛋白。与对照组比较，多奈哌齐组、LY组、SB组和SP各组sAPPα表达均有增加，sAPPβ表达均有减少，差异有统计学意义（P＜0.01）。各抑制剂组与多奈哌齐组比较，LY组sAPPα和sAPPβ表达无明显差异（P＞0.05），SB及SP组sAPPα表达明显降低，sAPPβ表达明显增加，，差异有统计学意义（（P＜0.01）。结论多奈哌齐能使APP的代谢向非淀粉样途径（α代谢途径）转变，其机制可能涉及JNK或P38信号通路，而与PI3-K通路无关。
[Abstract]:Objective to investigate the effect of Donepezil on non-amyloid metabolic pathway (伪 pathway) of amyloid precursor protein (APP) in PC12 cells and its possible signal mechanism. Methods PC12 cells cultured in vitro were divided into blank control group, Donepezil (20 渭 mol/L) group and LY294002 group (20 渭 mol/L) PI3-K inhibitor (LY group). Donepezil (20 渭 mol/L) P 38 inhibitor SB203580 group (SB group), Donepezil (20 渭 mol/L) JNK inhibitor SP600125 group (SP group). Each inhibitor group was given corresponding inhibitors 30 minutes before the addition of Donepezil. After 24 hours of culture, the levels of APP,ADAM10 and ADAM17 protein in the supernatant of cell culture were detected by Western blot and sAPP 伪 and sAPP/ 尾 in the supernatant were detected by ELISA method. Result 1. APP protein expression: there was no significant difference in the expression of APP protein between control group, Donepezil group, LY group, SB group and SP group (P > 0. 05). 2. ADAM10 and ADAM17 proteins: ADAM10 and ADAM17 proteins were detected by Western blot. Compared with the control group, the ADAM10 and ADAM17 of Donepezil group, LY group, SB group and SP group were increased significantly (P < 0. 05). The expression of ADAM10 and ADAM17 in LY group was not significantly different from that in Donepezil group (P > 0. 05), SB and SP group, P < 0. 05). 3. Expression of sAPP 伪 and sAPP 尾: sAPP 伪 and sAPP 尾 protein were detected by ELISA. Compared with the control group, the expression of sAPP 伪 in Donepezil group, LY group, SB group and SP group were all increased, and the expression of sAPP 尾 was decreased (P < 0. 01). The expression of sAPP 伪 and sAPP 尾 in LY group was not significantly different from that in Donepezil group (P > 0. 05), SB and SP group, sAPP 伪 expression was significantly decreased, sAPP 尾 expression was significantly increased (P < 0. 01). Conclusion Donepezil can change the metabolism of APP to non-amyloid pathway (伪 metabolic pathway), and its mechanism may be related to JNK or P38 signaling pathway, but not to PI3-K pathway.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R749.16

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