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目的观察脾气虚模型大鼠海马、下丘脑神经元线粒体形态学变化及其分裂情况,探讨脾气虚的发生机制。方法 20只SPF级雄性SD大鼠随机分为非脾气虚组和脾气虚模型组,每组10只。脾气虚模型组采用饮食失节+劳倦方法制脾气虚模型。透射电镜观察海马CA1区及下丘脑神经元线粒体的形态学变化;ELISA法检测海马和下丘脑组织ATP含量;Western blot法检测上述组织线粒体分裂因子(MFF)和线粒体分裂蛋白1(Fis1)的表达。结果非脾气虚组线粒体较丰富,形态基本正常,而脾气虚模型组线粒体数量减少,其中海马CA1区神经元线粒体出现肿胀、嵴减少,甚至空泡化,而下丘脑还存在较多线粒体自噬现象。与非脾气虚组比较,脾气虚模型组体重、进食量、饮水量、运动距离、站立次数以及前肢抓力均下降,海马和下丘脑神经元ATP含量降低,MFF及Fis1蛋白表达升高(P<0.05,P<0.01)。结论海马和下丘脑相关神经元线粒体功能低下及其促分裂成分表达增加与脾气虚的发生密切相关。
Objective To observe the changes of mitochondria and their mitosis in hippocampus and hypothalamus of rats with spleen deficiency syndrome and to explore the mechanism of spleen deficiency. Methods Twenty SPF male SD rats were randomly divided into non-spleen qi deficiency group and spleen qi deficiency model group, with 10 rats in each group. Spleen qi deficiency model group using the method of eating disorders + Luanxu spleen qi deficiency model. The morphological changes of mitochondria in hippocampal CA1 area and hypothalamus neurons were observed by transmission electron microscopy. The contents of ATP in hippocampus and hypothalamus were detected by ELISA. The expression of mitochondrial fission factor (MFF) and mitochondrial fission protein 1 . Results The mitochondria in the non-spleen qi deficiency group were more abundant and the morphology was basically normal. However, the number of mitochondria in the spleen qi deficiency model group was decreased. The mitochondria in the hippocampal CA1 region were swollen, the crest was reduced or even vacuolized, while more mitochondrial autophagy existed in the hypothalamus phenomenon. Compared with non-spleen qi deficiency group, body weight, food intake, water intake, exercise distance, standing times and forelimb grasping of spleen-qi deficiency group were decreased, ATP content of hippocampus and hypothalamus neurons decreased, and MFF and Fis1 protein expression increased <0.05, P <0.01). Conclusions The mitochondrial dysfunction and the expression of mitogenic components in hippocampus and hypothalamus related neurons are closely related to the occurrence of spleen deficiency.