针刺阳明经穴调控FALS患者NAD(P)转氢酶表达的研究
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摘要
目的:观察针刺阳明经穴后,家族性肌萎缩侧索硬化症(FALS)患者骨间肌中NAD(P)转氢酶的差异表达,揭示针刺阳明经穴治疗FALS的分子机制。方法:同一个家系患病组(4例)与正常组(5人),针刺穴位:双侧肩髃、曲池、合谷、伏兔、足三里、解溪,每日针刺1次,留针30min,每10min行针1次,留针期间共行针2次,10次为1个疗程,1个疗程后休息2d继续下1个疗程,共针刺3个疗程。治疗前后分别提取两组(9人)的骨间肌组织,运用同位素标记相对和绝对定量(iTRAQ)技术,运用Gene Ontology(GO)分析细胞成分,生物过程功能注释对NAD(P)转氢酶分析。结果:NAD(P)转氢酶表达在患病组治疗前后有显著性差异(P<0.01),并且表达呈上调。正常组治疗前后表达无差异。两组治疗前后比较表达均无显著差异。结论:针刺阳明经穴对FALS患者效应机制可能是通过对NAD(P)转氢酶的调控影响线粒体功能,抑制细胞凋亡,起到治疗作用。
Objective: To observe the different expression of NAD(P) transhydrogenase in the interosseous muscle in patients with FALS after acupuncturing yangming channel point, and to reveal the molecular mechanism of the treatment of familial amyotrophic lateral sclerosis by acupuncture yangming channel point. Methods: The diseased group(4 cases) and the control group(5 cases) which come from the same family. Acupuncture points: bilateral Jianyu(LI15), Quchi(LI11), Hegu(LI4),Futu(S32), Zusanli(S36), Jiexi(S41). Treatment: acupuncture once a day, retain the needle for 30 mins, manipulate the needle per 10 minutes and twice for each period of retaining the needle, 10 times as a course of treatment, 2-day rest after each treatment course. After that, begin the next course. Three courses are performed. Respectively extraction before and after the treatment between two groups(9) of interosseus tissue by using the technology of isotope-labeling method and absolute quantification(iTRAQ) using the Gene Ontology(GO) analysis the components of cell. Analysis the NAD(P) transhydrogenase by biological process fanctional annotation. Results: NAD(P) transhydrogenase expression of diseased group was significantly different(P<0.01)before and after treatment, and the result shows that the expression was up-regulated. However, there was no difference in the control group before and after treatment. There was no difference in protein expression before and after treatment in diseased and normal group. Conclusion: The effect mechanism of acupuncturing yangming channel point for patients with familial amyotrophic lateral sclerosis maybe through the regulation of NAD(P) transhydrogenase affect mitochondrial function, inhibiting cell apoptosis,playing a therapeutic role.
Objective: To observe the different expression of NAD(P) transhydrogenase in the interosseous muscle in patients with FALS after acupuncturing yangming channel point, and to reveal the molecular mechanism of the treatment of familial amyotrophic lateral sclerosis by acupuncture yangming channel point. Methods: The diseased group(4 cases) and the control group(5 cases) which come from the same family. Acupuncture points: bilateral Jianyu(LI15), Quchi(LI11), Hegu(LI4),Futu(S32), Zusanli(S36), Jiexi(S41). Treatment: acupuncture once a day, retain the needle for 30 mins, manipulate the needle per 10 minutes and twice for each period of retaining the needle, 10 times as a course of treatment, 2-day rest after each treatment course. After that, begin the next course. Three courses are performed. Respectively extraction before and after the treatment between two groups(9) of interosseus tissue by using the technology of isotope-labeling method and absolute quantification(iTRAQ) using the Gene Ontology(GO) analysis the components of cell. Analysis the NAD(P) transhydrogenase by biological process fanctional annotation. Results: NAD(P) transhydrogenase expression of diseased group was significantly different(P<0.01)before and after treatment, and the result shows that the expression was up-regulated. However, there was no difference in the control group before and after treatment. There was no difference in protein expression before and after treatment in diseased and normal group. Conclusion: The effect mechanism of acupuncturing yangming channel point for patients with familial amyotrophic lateral sclerosis maybe through the regulation of NAD(P) transhydrogenase affect mitochondrial function, inhibiting cell apoptosis,playing a therapeutic role.
引文
[1]吴珍元,黄英如,冼华.等.电针对失神经骨骼肌萎缩及纤维化的影响.中国康复医学杂志,2016,31(2):177-182
[2]Martin L J.The mitochondrial permeability transition pore:amolecular target for amyotrophic lateral sclerosis therapy.BiochimBiophys,2010,1802(1):186-197
[3]Lambeth J D,Kawahara T,Diebold B.Regulation of Nox andDuox enzymatic activity and expression.Free Radic Biol Med,2007,43(3):319-331
[4]Abdulwahid A I,Ahmad K H.Environmental toxins and Parkinson’sdisease:Putative roles of impaired electron transport chain andoxidative stress.Toxicol Ind Health,2010,26(2):121-128
[5]MagranéJ,Manfredi G.Mitochondrial function,morphology,andaxonal transport in amyotrophic lateral sclerosis.Antioxid RedoxSignal,2009,11(7):1615-1626
[6]李玲,黄银兰,马腾.国内外肌萎缩侧索硬化症发病相关蛋白研究进展.中风与神经疾病杂志,2014,10(31):953-955
[7]Hiqqins C M,Junq C,Xu Z.ALS-associated mutant SOD1G93Acauses mitochondrial vacuolation by expansion of theintermembrane space and by involvement of SOD1 aggregation andperoxisomes.BMC Neurosci,2003,15(4):16
[8]Hirano M,Angelini C,Montaqna P,et al.Amyotrophic lateralsclerosis with ragged-red fibers.Arch Neurol,2008,65(3):403-406
[9]Du H,Yan S S.Mitochondrial permeability transition pore inAlzheimer’s disease:Cyclophilin D and amyloid beta.BiochimBiophys Acta,2010,1802(1):198-204
[10]殷飞.肌萎缩侧索硬化与线粒体功能障碍的关系.长春:吉林大学,2007
[11]刘锐,黄慧玲.神经系统疾病线粒体蛋白质组学研究进展.天津医药,2007,35(11):875-878
[12]Wiedemann F R,Winkler K,Kuznetsov A V,et al.Impairmentof mitochondrial function in skeletal muscle of patients withamyotrophic lateral sclerosis.J Neurol Sci,1998,156(1):65-72
[13]Dupuis L,Gonzalez de Aguilar J L,Oudart H,et al.Mitochondria inamyotrophic lateral sclerosis:A trigger and a target.NeurodegenerDis,2004,1(6):245-254
[14]Stefano L,Massimo P,Alberto F,et al.Dynamic NAD(P)Hpost-synaptic autofluorescence signals for the assessment ofmitochondrial function in a neurodegenerative disease:Monitoringthe primary motor cortex of G93A mice,an amyotrophic lateralsclerosis model,Mitochondrion,2010,10(2):108-114
[15]SongW,SongY,KincaidB,etal.MutantSOD1G93Atriggers mitochondrial fragmentation in spinal cord motorneurons:neuroprotection by SIRT3 and PGC-1α.Neurobiol Dis,2013,51:72-81
[16]Magnifico S,Saias L,Deleglise B,et al.NAD+acts on mitochondrialSir T3 to prevent axonal caspase activation and axonaldegeneration.FASEB J,2013,27(12):4712-4722
[17]李达,伦永志,周士胜.NAD+/NADH代谢机制研究进展.生物技术通讯,2010,21(1):98-102
[2]Martin L J.The mitochondrial permeability transition pore:amolecular target for amyotrophic lateral sclerosis therapy.BiochimBiophys,2010,1802(1):186-197
[3]Lambeth J D,Kawahara T,Diebold B.Regulation of Nox andDuox enzymatic activity and expression.Free Radic Biol Med,2007,43(3):319-331
[4]Abdulwahid A I,Ahmad K H.Environmental toxins and Parkinson’sdisease:Putative roles of impaired electron transport chain andoxidative stress.Toxicol Ind Health,2010,26(2):121-128
[5]MagranéJ,Manfredi G.Mitochondrial function,morphology,andaxonal transport in amyotrophic lateral sclerosis.Antioxid RedoxSignal,2009,11(7):1615-1626
[6]李玲,黄银兰,马腾.国内外肌萎缩侧索硬化症发病相关蛋白研究进展.中风与神经疾病杂志,2014,10(31):953-955
[7]Hiqqins C M,Junq C,Xu Z.ALS-associated mutant SOD1G93Acauses mitochondrial vacuolation by expansion of theintermembrane space and by involvement of SOD1 aggregation andperoxisomes.BMC Neurosci,2003,15(4):16
[8]Hirano M,Angelini C,Montaqna P,et al.Amyotrophic lateralsclerosis with ragged-red fibers.Arch Neurol,2008,65(3):403-406
[9]Du H,Yan S S.Mitochondrial permeability transition pore inAlzheimer’s disease:Cyclophilin D and amyloid beta.BiochimBiophys Acta,2010,1802(1):198-204
[10]殷飞.肌萎缩侧索硬化与线粒体功能障碍的关系.长春:吉林大学,2007
[11]刘锐,黄慧玲.神经系统疾病线粒体蛋白质组学研究进展.天津医药,2007,35(11):875-878
[12]Wiedemann F R,Winkler K,Kuznetsov A V,et al.Impairmentof mitochondrial function in skeletal muscle of patients withamyotrophic lateral sclerosis.J Neurol Sci,1998,156(1):65-72
[13]Dupuis L,Gonzalez de Aguilar J L,Oudart H,et al.Mitochondria inamyotrophic lateral sclerosis:A trigger and a target.NeurodegenerDis,2004,1(6):245-254
[14]Stefano L,Massimo P,Alberto F,et al.Dynamic NAD(P)Hpost-synaptic autofluorescence signals for the assessment ofmitochondrial function in a neurodegenerative disease:Monitoringthe primary motor cortex of G93A mice,an amyotrophic lateralsclerosis model,Mitochondrion,2010,10(2):108-114
[15]SongW,SongY,KincaidB,etal.MutantSOD1G93Atriggers mitochondrial fragmentation in spinal cord motorneurons:neuroprotection by SIRT3 and PGC-1α.Neurobiol Dis,2013,51:72-81
[16]Magnifico S,Saias L,Deleglise B,et al.NAD+acts on mitochondrialSir T3 to prevent axonal caspase activation and axonaldegeneration.FASEB J,2013,27(12):4712-4722
[17]李达,伦永志,周士胜.NAD+/NADH代谢机制研究进展.生物技术通讯,2010,21(1):98-102