CAJ | 학술논문

目的探讨在阿尔茨海默病(AD)脑损伤中,β-淀粉样蛋白(Aβ)神经毒性对大鼠行为学、自噬相关蛋白Beclin-1和LC3-Ⅱ的影响及何首乌提取物二苯乙烯苷(TSG)的干预作用.方法选Wistar大鼠80只,随机数字表法分为对照组、假手术组、模型组、TSG组,各20只.采用立体定向仪下于海马部位注射微量Aβ1-42造模,Y电迷宫及Moms水迷宫检测行为学变化,反转录聚合酶链反应及Western blot法检测制模后第21天海马神经元内自噬相关蛋白Beclin-1和LC3 -Ⅱ的mRNA及蛋白表达变化.结果在模型组中,大鼠Y电迷宫躲避所需的电刺激次数增加,Morris水迷宫测试中潜伏期延长,游泳路程增加及穿越平台次数减少；Beclin-1和LC3-Ⅱ的mRNA及蛋白的表达一致,在21 d时,模型组Beclin-1蛋白(0.51±0.03),LC3-Ⅱ蛋白(0.68 ±0.04)与对照组(0.31±0.01、0.31±0.02)比较,差异有统计学意义(t=28.2843、37.0000,均P＜0.05)；TSG干预后,大鼠躲避所需的电刺激次数减少,潜伏期缩短,游泳路程缩短,穿越平台次数增加；Beclin-1和LC3-Ⅱ蛋白表达强度较模型组减弱,差异有统计学意义(t=9.8387、16.2698,均P＜0.05).结论海马神经元在受到Aβ刺激后,可上调自噬蛋白Beclin-1和LC3-Ⅱ因子表达,启动自噬通路；TSG可通过减轻内质网应激损害,下调Beclin-1和LC3-Ⅱ的表达来抵抗Aβ的神经毒性,改善大鼠行为学表现,发挥脑保护作用.
목적 탐토재아이자해묵병(AD)뇌손상중,β-정분양단백(Aβ)신경독성대대서행위학、자서상관단백Beclin-1화LC3-Ⅱ적영향급하수오제취물이분을희감(TSG)적간예작용.방법 선Wistar대서80지,수궤수자표법분위대조조、가수술조、모형조、TSG조,각20지.채용입체정향의하우해마부위주사미량Aβ1-42조모,Y전미궁급Moms수미궁검측행위학변화,반전록취합매련반응급Western blot법검측제모후제21천해마신경원내자서상관단백Beclin-1화LC3 -Ⅱ적mRNA급단백표체변화.결과 재모형조중,대서Y전미궁타피소수적전자격차수증가,Morris수미궁측시중잠복기연장,유영로정증가급천월평태차수감소；Beclin-1화LC3-Ⅱ적mRNA급단백적표체일치,재21 d시,모형조Beclin-1단백(0.51±0.03),LC3-Ⅱ단백(0.68 ±0.04)여대조조(0.31±0.01、0.31±0.02)비교,차이유통계학의의(t=28.2843、37.0000,균P＜0.05)；TSG간예후,대서타피소수적전자격차수감소,잠복기축단,유영로정축단,천월평태차수증가；Beclin-1화LC3-Ⅱ단백표체강도교모형조감약,차이유통계학의의(t=9.8387、16.2698,균P＜0.05).결론 해마신경원재수도Aβ자격후,가상조자서단백Beclin-1화LC3-Ⅱ인자표체,계동자서통로；TSG가통과감경내질망응격손해,하조Beclin-1화LC3-Ⅱ적표체래저항Aβ적신경독성,개선대서행위학표현,발휘뇌보호작용.
Objective To observe the effect of tetrahydroxy stilbene glucoside (TSG) on the behavior on rat model and the expressions of autophagy-associated protein Beclin-1 and LC3- Ⅱ induced by Aβ1-42 Methods Eighty rats were equally randomized into 4 groups (n =20):The control group,the sham operated group,the model group and the TSG group.The behavior of rats was measured by using Y-maze and Morris water maze.The expression of Beclin-1 and LC3- Ⅱ in rats hippocampus was detected by Western blot and RT-PCR at the time points.Results The number of electric-stimulus in hippocampus significantly increased and the Morris water maze test showed that the escape latency prolonged,swimming distance increased and the times of crossing the exact former platform location decreased both in the model and TSG groups after 21 days compared with those in control group.The mRNAs and protein expressions of Beclin-1 (0.51 ±0.03)and LC3-Ⅱ (0.68 ± 0.04) in model group were higher than that in control group (0.31 ± 0.01,0.31 ± 0.02) at that time point ( Beclin-1:t =28.2843,P ＜ 0.05 ; LC3- Ⅱ :t =37.0000,P ＜0.05).Compared to model group,the expression of the Beclin-1 and LC3- Ⅱ was decreased at 21 d in TSG group (Beclin-1:t =9.8387,P ＜ 0.05 ; LC3- Ⅱ :t =16.2698,P ＜ 0.05 ).Conclusions Autophagy self-regulated system is started through the increased expressions of Beclin-1 and LC3- Ⅱ after Aβ deposition in rats,so as to attenuate cerebral injury caused by Aβ neurotoxicity.Autophagy pathway is possible one of the mechanisms in Aβ neurotoxic injury. Tetrahydroxy stilbene glucoside from polygonum multiflorum has protective effect on it.