滋肾益髓方对帕金森病小鼠分子伴侣介导的自噬途径的影响
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摘要
目的:通过观察帕金森病(PD)小鼠脑内组织蛋白酶D(Cath D)、热休克蛋白70(HSP70)的表达探讨中药滋肾益髓方对分子伴侣介导的自噬途径(CMA)的影响及其治疗PD可能机制。方法:将C57BL/6雄性小鼠随机分为正常组、模型组、西药组和中药组。鱼藤酮灌胃法建立PD小鼠模型,给药组在造模后采取不同治疗措施。自主活动测试法观察小鼠行为学;免疫组织化学技术检测小鼠中脑黑质Cath D、HSP70表达。结果:经治疗后中药组及西药组小鼠自主活动次数均较模型组明显增高(P<0.05)。模型组小鼠黑质区Cath D、HSP70表达较空白组明显减少(P<0.05),中药组及西药组小鼠黑质区Cath D、HSP70表达较模型组明显增高(P<0.05)。结论:中药滋肾益髓方可能通过增强CMA途径达到治疗PD的目的。
Objective: By observation the expression of Cath D and HSP70 in the brain of mice with Parkinson's disease(PD), to investigate the possible mechanism of Companion-mediated autophagy(CMA) for the treatment of PD. Methods: C57 BL/6 male mice were randomly divided into control group, model group, Madopar group and TCM group. The model of PD mice was established by the method of rotenone, and the latter two groups were treated with different treatment measures after modeling. The behavior of mice was observed by the autonomous activity test. Immunohistochemical technique was used to detect the expression of Cath D and HSP70 in substantia nigra of mice. Results: After treatment, the activity of the mice in TCM group and Madopar group was significantly higher than that in model group(P<0.05). The expression of Cath D and HSP70 in the substantia nigra of model group was significantly lower than that in control group(P<0.05). The expression of Cath D and HSP70 in the substantia nigra of TCM group and Madopar group was significantly higher than that in model group(P<0.05). Conclusion: Zishen Yisui Formula could achieve the purpose of treating PD by enhancing the CMA pathway.
Objective: By observation the expression of Cath D and HSP70 in the brain of mice with Parkinson's disease(PD), to investigate the possible mechanism of Companion-mediated autophagy(CMA) for the treatment of PD. Methods: C57 BL/6 male mice were randomly divided into control group, model group, Madopar group and TCM group. The model of PD mice was established by the method of rotenone, and the latter two groups were treated with different treatment measures after modeling. The behavior of mice was observed by the autonomous activity test. Immunohistochemical technique was used to detect the expression of Cath D and HSP70 in substantia nigra of mice. Results: After treatment, the activity of the mice in TCM group and Madopar group was significantly higher than that in model group(P<0.05). The expression of Cath D and HSP70 in the substantia nigra of model group was significantly lower than that in control group(P<0.05). The expression of Cath D and HSP70 in the substantia nigra of TCM group and Madopar group was significantly higher than that in model group(P<0.05). Conclusion: Zishen Yisui Formula could achieve the purpose of treating PD by enhancing the CMA pathway.
引文
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[12]Hinault M P,Cuendet A F,Mattoo R U,et al.Stable alpha-synuclein oligomers strongly inhibit chaperone activity of the Hsp70system by weak interactions with J-domain co-chaperones.Biol Chem,2010,285(49):38173-38182
[13]Dedmon M M,Christodoulou J,Wilson M R,et al.Heat shockprotein70 inhibits alpha-synuclein fibril formation via preferentialbinding to prefibrillar species.J Biol Chem,2005,280(15):14733-14740
[14]Huang C,Cheng H,Hao S,et al.Heat shock protein 70 inhibits alpha-synuclein fibril formation via interactions with diverse intermediates.J Mol Biol,2006,364:323-336
[15]陈宏志,李静蔚,何建成.基于临床病案文献的帕金森病中医基本证候研究.中华中医药杂志,2016,31(8):3216-3219
[2]Braak H,Del Tredici K,Rub U,et al.Staging of brain pathology related to sporadic Parkinson’s disease.Neurobiol Aging,2003,24:197-211
[3]Kuo Y M,Li Z,Jiao Y,et al.Extensive enteric nervous system abnormalities in mice transgenic for artificial chromosomes containing Parkinson disease-associated alpha-synuclein gene mutations precede central nervous system changes.Hum Mol Genet,2010,19(9):1633-1650
[4]陈路,陈志刚,侯月.滋肾益髓方治疗帕金森病运动及非运动症状的疗效观察.北京中医药大学学报,2014,10(3):57-59
[5]韦一佛.滋肾益髓方对帕金森病模型小鼠脑、胃、肠α-synuclein的影响.北京:北京中医药大学,2015
[6]Sevlever D,Jiang P,Yen S H.Cathepsin D is the main lysosomal enzyme involved in the degradation of alpha-synuclein and generation of its carboxy-terminally truncated species.Biochemistry,2009,47:9678-9687
[7]Pan-Montojo F,Anichtchik O,Funk R H,et al.Progression of Parkinsn’s disease pathology is reproduced by intragastric administration of rotenone in mice.PLo S One,2010,5(1):e8762
[8]Kelvin Cuk,Victoria Kehm.Paα-synologicalα-syn transmission initiates parkinson-like neurodegeneration in non-transgenic mice.Science,2012,338(6109):949-953
[9]Kalia L V,Kalia S K,Mclean P J,et al.alpha-Synuclein oligomers and clinical implications for Parkinson disease.Ann Neurol,2013,73(2):155-169
[10]Giehm L,Svergun D I,Otzen D E,et al.Low-resolution structure of a vesicle disrupting&alpha synuclein oligomer that accumulates during fibrillation.Proc Natl Acad Sci USA,2011,108(8):3246-3251
[11]Cullen V,Lindfors M,Ng J,et al.Cathepsin D expression level affects alpha-synuclein processing,aggregation,and toxicity in vivo.Mol Brain,2009,2:5
[12]Hinault M P,Cuendet A F,Mattoo R U,et al.Stable alpha-synuclein oligomers strongly inhibit chaperone activity of the Hsp70system by weak interactions with J-domain co-chaperones.Biol Chem,2010,285(49):38173-38182
[13]Dedmon M M,Christodoulou J,Wilson M R,et al.Heat shockprotein70 inhibits alpha-synuclein fibril formation via preferentialbinding to prefibrillar species.J Biol Chem,2005,280(15):14733-14740
[14]Huang C,Cheng H,Hao S,et al.Heat shock protein 70 inhibits alpha-synuclein fibril formation via interactions with diverse intermediates.J Mol Biol,2006,364:323-336
[15]陈宏志,李静蔚,何建成.基于临床病案文献的帕金森病中医基本证候研究.中华中医药杂志,2016,31(8):3216-3219