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
|
摘要
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.
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.
引文
[1]Holtmann G,Talley N J.The stomach-brain axis.Best Pract Res Clin Gastroenterol,2014,28(6):967-979
[2]Suyama S,Takano E,Iwasaki Y,et al.Roles and functional interplay of the gut, brain stem,hypothalamus and limbic system in regulation of feeding.Nihon Rinsho,2009,67(2):277-286
[3]刘垒,冯杜,朱玉山,等.线粒体自噬的研究进展.中国细胞生物学学报,2012,34(10):959-966
[4]Liesa M,Shirihai O S.Mitochondrial dynamics in the regulation of nutrient utilization and energy expenditure.Cell Metab,2013,17(4):491-506
[5]Dhingra R,Kirshenbaum L A.Regulation of mitochondrial dynamics and cell fate.Circ J,2014,78(4):803-810
[6]刘文俊,陈伟仁,宋囡,等.脾气虚模型大鼠海马与下丘脑神经元线粒体分裂因子和线粒体分裂蛋白1的表达.中国中西医结合杂志,2017,37(11):1356-1360
[7]游艳婷,金丽琴.中医脾虚证与线粒体的相关性.温州医科大学学报,2015,45(10):774-778
[8]侯丽颖,刘友章,贺松其,等.中医脾与线粒体功能的相关性探讨.上海中医药杂志,2008,42(7):3-4
[9]郑敏麟,阮诗玮.中医藏象实质细胞生物学假说之一——“脾”与线粒体.中国中医基础医学杂志,2002,8(5):10-12
[10]刘旭东,刘文俊,孙大宇,等.脾气虚证模型大鼠神疲乏力的客观化评价.中华中医药杂志,2015,30(3):699-701
[11]Casanova J E.Bacterial Autophagy:Offense and Defense at the Host-Pathogen Interface.Cell Mol Gastroenterol Hepatol,2017,4(2):237-243
[12]Arakawa S,Honda S,Yamaguchi H,et al.Molecular mechanisms and physiological roles of Atg5/Atg7-independent alternative autophagy.Proc Jpn Acad Ser B Phys Biol Sci,2017,93(6):378-385
[13]姜辉,秦秀娟,万磊,等.五味温通除痹胶囊对佐剂性关节炎大鼠自噬蛋白Beclin-1、LC3-Ⅱ表达的影响.中成药,2017,39(8):1566-1572
[14]Gào X,Sch?ttker B.Reduction-oxidation pathways involved in cancer development:a systematic review of literature reviews.Oncotarget,2017,8(31):51888-51906
[15]Rottenberg H,Hoek J B.The path from mitochondrial ROS to aging runs through the mitochondrial permeability transition pore.Aging Cell,2017,16(5):943-955
[16]Nguyen T N,Padman B S,Lazarou M.Deciphering the Molecular Signals of PINK1/Parkin Mitophagy.Trends Cell Biol,2016,26(10):733-744
[17]Novak I.Mitophagy:A complex mechanism of mitochondrial removal.Antioxid Redox Signal,2012,17(5):794-802
[18]柏杖勇,李清华.PINK1/parkin,线粒体自噬与帕金森病.中国老年学杂志,2014,34(9):2609-2613
[2]Suyama S,Takano E,Iwasaki Y,et al.Roles and functional interplay of the gut, brain stem,hypothalamus and limbic system in regulation of feeding.Nihon Rinsho,2009,67(2):277-286
[3]刘垒,冯杜,朱玉山,等.线粒体自噬的研究进展.中国细胞生物学学报,2012,34(10):959-966
[4]Liesa M,Shirihai O S.Mitochondrial dynamics in the regulation of nutrient utilization and energy expenditure.Cell Metab,2013,17(4):491-506
[5]Dhingra R,Kirshenbaum L A.Regulation of mitochondrial dynamics and cell fate.Circ J,2014,78(4):803-810
[6]刘文俊,陈伟仁,宋囡,等.脾气虚模型大鼠海马与下丘脑神经元线粒体分裂因子和线粒体分裂蛋白1的表达.中国中西医结合杂志,2017,37(11):1356-1360
[7]游艳婷,金丽琴.中医脾虚证与线粒体的相关性.温州医科大学学报,2015,45(10):774-778
[8]侯丽颖,刘友章,贺松其,等.中医脾与线粒体功能的相关性探讨.上海中医药杂志,2008,42(7):3-4
[9]郑敏麟,阮诗玮.中医藏象实质细胞生物学假说之一——“脾”与线粒体.中国中医基础医学杂志,2002,8(5):10-12
[10]刘旭东,刘文俊,孙大宇,等.脾气虚证模型大鼠神疲乏力的客观化评价.中华中医药杂志,2015,30(3):699-701
[11]Casanova J E.Bacterial Autophagy:Offense and Defense at the Host-Pathogen Interface.Cell Mol Gastroenterol Hepatol,2017,4(2):237-243
[12]Arakawa S,Honda S,Yamaguchi H,et al.Molecular mechanisms and physiological roles of Atg5/Atg7-independent alternative autophagy.Proc Jpn Acad Ser B Phys Biol Sci,2017,93(6):378-385
[13]姜辉,秦秀娟,万磊,等.五味温通除痹胶囊对佐剂性关节炎大鼠自噬蛋白Beclin-1、LC3-Ⅱ表达的影响.中成药,2017,39(8):1566-1572
[14]Gào X,Sch?ttker B.Reduction-oxidation pathways involved in cancer development:a systematic review of literature reviews.Oncotarget,2017,8(31):51888-51906
[15]Rottenberg H,Hoek J B.The path from mitochondrial ROS to aging runs through the mitochondrial permeability transition pore.Aging Cell,2017,16(5):943-955
[16]Nguyen T N,Padman B S,Lazarou M.Deciphering the Molecular Signals of PINK1/Parkin Mitophagy.Trends Cell Biol,2016,26(10):733-744
[17]Novak I.Mitophagy:A complex mechanism of mitochondrial removal.Antioxid Redox Signal,2012,17(5):794-802
[18]柏杖勇,李清华.PINK1/parkin,线粒体自噬与帕金森病.中国老年学杂志,2014,34(9):2609-2613