慢性乙醇中毒小鼠脑神经细胞IP3R1表达的变化

Chronic ethanol poisoning induced the changes of IP3R1 in mouse cerebral cortex neurons

目的研究慢性乙醇中毒引起小鼠脑神经细胞Ⅰ型1,4,5-三磷酸肌醇受体(IP3R1)表达的变化。方法 40只小鼠随机分为90d、180d组,各组再分为正常对照组、10%、20%、30%乙醇组,每组5只,乙醇组给予相应浓度乙醇饮用至相应时间;取各组小鼠脑组织,分别采用免疫组化和Western blot方法检测脑皮质神经细胞IP3R1表达的变化;SPSS 13.0软件对实验数据进行单因素方差分析。结果正常IP3R1免疫组化染色物分布于神经细胞胞浆内。90d组随乙醇浓度的增加,IP3R1免疫组化阳性细胞数和阳性细胞率逐步增加,组间比较,差异显著(P<0.05);180d组中10%、20%乙醇组阳性细胞数和阳性细胞率逐步增加,组间比较,差异显著(P<0.05),而30%乙醇组阳性细胞数和阳性细胞率反而减少,且低于90d组的相同浓度组(P<0.05)。Western blot与免疫组化检测结果基本一致。结论慢性乙醇中毒可引起小鼠大脑皮质神经细胞IP3R1表达增加,而高浓度(30%)、长时间(180d)乙醇使IP3R1表达降低,可能与神经细胞变性、坏死、数目减少有关。

Objective To investigate the changes of type 1 inositol 1,4,5-trisphosphate receptor（ IP3R1） in mouse cerebral cortex neurons with chronic ethanol exposure. Methods 40 mice were randomly divided into 8 groups and given water,10%,20% and 30% ethanol（ v / v） for drinking during 24 hour for 90 day and 180 day periods respectively. The chronic ethanol poisoning-induced changes of IP3R1 in mouse cerebral cortex neurons were detected by immunohistochemical staining and western blotting. The data were statistically analyzed by SPSS13. 0 software. Results The IP3R1 immunoreactivity was detected in mouse cerebral cortex neurons. The number and ratio of IP3R1 positive cells in mouse cerebral cortex neurons increased gradually in each 90d ethanol groups and in 180d 10%,20% ethanol groups. The average optical densities of IP3R1 protein bands detected by western blot increased correspondingly. There were statistically significant differences（ P ＆lt; 0. 05） between the water and ethanol groups（ P ＆lt; 0. 05）,as well as between two neighboring ethanol groups. But there was a significant decrease（ P ＆lt; 0. 05） in number and ratio of IP3R1 positive cells and average optical density of IP3R1 protein bands in 180d 30% ethanol group,as compared with the 180d 10% 、20% and 90d 30% ethanol groups,respectively. Conclusion Chronic ethanol poisoning increased the expression of IP3R1 in mouse cerebral cortex neurons. But the high-concentration ethanol and long-peroid poisoning decreased the expression of IP3R1,which maybe related withethanol-induced neuro-degeneration,necrosis and cell loss.