Aβ25-35通过钙超载导致HT22神经细胞Per2异常表达
发布时间：2018-10-23 21:16
【摘要】：目的探讨钙超载在Aβ25-35引起HT22细胞Per2表达变化中的作用。方法体内研究将C57BL/6小鼠双侧海马注射Aβ25-35(300μmol/kg)和无菌水后,利用跑轮行为学实验评估小鼠的昼夜节律。体外研究将HT22细胞分为对照组、Aβ25-35处理组、Nim+Aβ25-35处理组。以MTT法检测细胞存活率,实时荧光定量PCR检测per2 mRNA表达水平,Western Blotting检测PER2蛋白相对表达量,免疫荧光染色观察PER2蛋白原位表达情况,流式细胞术检测胞内钙离子浓度。结果体内研究表明Aβ25-35导致C57BL/6小鼠在跑轮行为学实验中表现出明显的昼夜节律紊乱。离体研究发现,5、10、15、20、25、30μmol/L Aβ25-35剂量组细胞存活率较对照组显著降低(P0.05)。经20μmol/L Aβ25-35处理后,per2 mRNA及PER2蛋白水平在CT16较对照组显著降低(P0.05)。经钙离子拮抗剂尼莫地平预处理后,由Aβ25-35在CT16所致per2 mRNA及PER2蛋白水平的降低均显著升高(P0.05)。结论钙超载在Aβ25-35引起HT22细胞Per2异常表达中发挥重要作用。
[Abstract]:Objective to investigate the role of calcium overload in the changes of Per2 expression in HT22 cells induced by A 尾 25-35. Methods A 尾 25-35 (300 渭 mol/kg) and aseptic water were injected into the hippocampus of C57BL/6 mice in vivo. HT22 cells were divided into two groups in vitro: control group, A 尾 25-35 group, Nim A 尾 25-35 group. The cell survival rate was detected by MTT method, the per2 mRNA expression level was detected by real-time fluorescence quantitative PCR, the relative expression of PER2 protein was detected by, Western Blotting, the expression of PER2 protein in situ was observed by immunofluorescence staining, and the intracellular calcium concentration was detected by flow cytometry. Results in vivo studies showed that A 尾 25-35 resulted in obvious circadian rhythm disorder in C57BL/6 mice. In vitro study, the cell survival rate of the group at the dose of 10 ~ 10 ~ 15A 尾 25 ~ (35) 渭 mol/L A 尾 25 ~ 35 was significantly lower than that of the control group (P0.05). After treated with 20 渭 mol/L A 尾 25-35, the levels of per2 mRNA and PER2 protein in CT16 were significantly lower than those in control group (P0.05). Pretreatment with Nimodipine, a calcium antagonist, significantly increased the levels of per2 mRNA and PER2 protein in CT16 induced by A 尾 25-35 (P0.05). Conclusion calcium overload may play an important role in the abnormal expression of Per2 in HT22 cells induced by A 尾 25-35.
【作者单位】： 山西医科大学基础医学院病理教研室;山西医科大学基础医学院形态学实验室;
【基金】：国家自然科学基金资助项目(81471343) 山西医科大学科技创新基金资助项目(01201307)
【分类号】：R749.16
，
本文编号：2290482
[Abstract]:Objective to investigate the role of calcium overload in the changes of Per2 expression in HT22 cells induced by A 尾 25-35. Methods A 尾 25-35 (300 渭 mol/kg) and aseptic water were injected into the hippocampus of C57BL/6 mice in vivo. HT22 cells were divided into two groups in vitro: control group, A 尾 25-35 group, Nim A 尾 25-35 group. The cell survival rate was detected by MTT method, the per2 mRNA expression level was detected by real-time fluorescence quantitative PCR, the relative expression of PER2 protein was detected by, Western Blotting, the expression of PER2 protein in situ was observed by immunofluorescence staining, and the intracellular calcium concentration was detected by flow cytometry. Results in vivo studies showed that A 尾 25-35 resulted in obvious circadian rhythm disorder in C57BL/6 mice. In vitro study, the cell survival rate of the group at the dose of 10 ~ 10 ~ 15A 尾 25 ~ (35) 渭 mol/L A 尾 25 ~ 35 was significantly lower than that of the control group (P0.05). After treated with 20 渭 mol/L A 尾 25-35, the levels of per2 mRNA and PER2 protein in CT16 were significantly lower than those in control group (P0.05). Pretreatment with Nimodipine, a calcium antagonist, significantly increased the levels of per2 mRNA and PER2 protein in CT16 induced by A 尾 25-35 (P0.05). Conclusion calcium overload may play an important role in the abnormal expression of Per2 in HT22 cells induced by A 尾 25-35.
【作者单位】： 山西医科大学基础医学院病理教研室;山西医科大学基础医学院形态学实验室;
【基金】：国家自然科学基金资助项目(81471343) 山西医科大学科技创新基金资助项目(01201307)
【分类号】：R749.16
，
本文编号：2290482
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