脾气虚大鼠海马神经元线粒体自噬水平及PINK1/Parkin途径的研究

Study on mitophagy and PINK1/Parkin pathway of hippocampal neurons of spleen qi deficiency rats

目的:通过观察脾气虚大鼠海马神经元线粒体自噬水平及PTEN诱导激酶1(PINK1)/Parkin途径的变化,从线粒体方面研究脾气虚的发生机制。方法:16只雄性SD大鼠随机分为对照组和脾气虚模型组(模型组),每组8只;透射电镜观察海马神经元线粒体形态学变化;比色法检测海马组织ATP含量;Western blot法检测海马神经元微管相关蛋白1轻链3(LC3)-Ⅰ和Ⅱ、PINK1及Parkin蛋白表达。结果:与对照组比较,模型组海马神经元线粒体数量减少、空泡化,但未见明显自噬现象;ATP含量显著减少(P<0.01),LC3-Ⅱ/Ⅰ比值变小(P<0.05),LC3-Ⅱ、PINK1和Parkin蛋白表达显著下调(P<0.05)。结论:脾气虚的发生可能与PINK1/Parkin途径抑制所致的海马神经元线粒体自噬水平下降有关。

Objective: To observe the degree of mitophagy and change of PINK1/Parkin pathway of rat hippocampal neurons in spleen qi deficiency, exploring the pathogenesis of spleen qi deficiency in mitochondria. Methods: Sixteen male SD rats were randomly divided into the control group and spleen qi deficiency group(model group), 8 in each group;Transmission electron microscopy was used to observe mitochondrial morphology in hippocampal neurons;hippocampal ATP levels ELISA was measured by ELISA;the expressions of light chain 3(LC3)-Ⅰ and Ⅱ, PTEN induced putative kinase 1(PINK1) and Parkin in hippocampus were measured by Western blotting. Results: Compared with those in the control, mitochondria in hippocampal neurons was less in number and swollen, ATP level was reduced significantly(P<0.01), the ratio of LC3-Ⅱ/Ⅰ was decreased markedly(P<0.05), and expressions of LC3-Ⅱ, PINK1 and Parkin were all reduced significantly(P<0.05). Conclusion: The pathogenesis of spleen qi deficiency might be related to the attenuated mitophagy caused by the inhibition of PINK1/Parkin pathway.