摘要
目的:观察胃泌酸调节素类似物D-Ser2-Oxyntomodulin(Oxy)是否可以改善β淀粉样蛋白31-35(Aβ31-35)所致的小鼠昼夜节律紊乱,探讨Oxy改善Aβ31-35所致昼夜节律紊乱的细胞分子机制。方法:(1)选取6~8周龄C57BL/6雄性小鼠进行跑轮行为学实验,分析Oxy改善Aβ31-35所致小鼠昼夜节律紊乱的作用。(2)选取小鼠海马HT22神经细胞,CCK8法检测细胞存活率,Real-time PCR检测不同时间点钟基因Bmal1和Per2 mRNA的变化趋势,Western Blot分别检测CT20时Bmal1蛋白和CT16时Per2蛋白的表达情况。结果:(1)Aβ31-35引起小鼠昼夜节律紊乱,与对照组相比,自由运转周期显著延长。Oxy预处理再给予Aβ31-35后小鼠的昼夜节律紊乱情况有所改善,与Aβ单独给予相比,自由运转周期显著降低。(2)Aβ31-35引起HT22细胞存活率显著降低,Oxy预处理后有效逆转Aβ31-35诱导的细胞存活率下降,且具有剂量依赖性。(3)经Aβ31-35处理后,HT22细胞的Bmal1和Per2 mRNA水平分别在CT20和CT16较对照组明显降低;而Oxy预处理组Bmal1和Per2的异常表达均得到显著改善。(4)与对照组相比,Aβ31-35组CT20时Bmal1和CT16时Per2蛋白水平显著降低;而与Aβ31-35组相比,Oxy预处理组Bmal1和Per2蛋白水平有所升高。结论:Oxy可能通过调整Bmal1和Per2的表达改善Aβ31-35所致C57BL/6小鼠昼夜节律紊乱。
Objective： To observe the effect of D-Ser2-Oxyntomodulin（ Oxy） on the circadian rhythm disorder induced by amyloid β-protein 31-35（ Aβ31-35） in mice,and investigate the cellular and molecular mechanisms of Oxy in improving circadian rhythm disorder induced by Aβ31-35. Methods：（ 1） 6 - 8 weeks old C57 BL/6 male mice were selected for wheel-running behavior test and analyze the effect of Oxy on the circadian rhythm disorder induced by Aβ31-35.（ 2） HT22 neural cells of mice hippocampus were chosen to reveal the cellular and molecular mechanisms of Oxy in improving circadian rhythm disorder induced by Aβ31-35. The cell viability was measured by CCK8,and Real-time PCR was used to detect the expression of clock genes Bmal1 and Per2. Western Blot was applied to examine the expression of Bmal1 protein at CT20 and Per2 protein at CT16. Results：（ 1） Compared with control group,Aβ31-35 disturbed the circadian rhythm which exhibited significantly longer free running period. Whereas the disruption was significantly relieved with pretreatment of Oxy compared with Aβ31-35 treatment,which manifested in significantly decreasedfree running period.（ 2） Aβ31-35 decreased survival rate of HT22 cells compared to control group,and Oxy can reduce the toxicity of Aβ31-35 by up-regulating the cell survival rate in a dose-dependent way.（ 3） Abnormal rhythmic expressions of Bmal1 and Per2 mRNA were induced by Aβ31-35 in HT22 cells which significantly decreased at CT20 and CT16 respectively compared with control group. Oxy pre-treatment can reverse the abnormal expression.（ 4） Aβ31-35 significantly decreased Bmal1 and Per2 protein expressions at CT20 and CT16,respectively. Pre-treatment with Oxy can significantly improve the expressions of Bmal1 and Per2 protein. Conclusion： Oxy may ameliorate Aβ31-35 induced circadian rhythm disorder by regulating Bmal1 and Per2 expressions.
作者
赵今
原媛
张蕊
王昌图
王丽
王晓晖
Zhao Jin;Yuan Yuan;Zhang Rui;Wang Changtu;Wang Li;Wang Xiaohui(Department of Pathology;Department of Morphology, Preclinical Medical Experiment Teaching Center, College of Basic Medicine, Shanxi Medical University, Taiyuan 030001, China)
出处
《神经解剖学杂志》
CAS
CSCD
北大核心
2018年第3期305-312,共8页
Chinese Journal of Neuroanatomy
基金
国家自然科学基金(81471343)
山西医科大学科技创新基金(01201307)
