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目的通过观察慢病毒介导的sh RNA沉默TRPV4基因对帕金森模型小鼠的影响,探讨TRPV4在帕金森病发展中的作用。方法慢病毒表达载体TRPV4 sh RNA感染小鼠黑质致密部,1周后腹腔注射1-甲基-4-苯基-1,2,3,6-四氢吡啶(30 mg·kg-1)造模。小鼠随机分为5组,分别为正常对照组、MPTP组(模型组)、Scrimble sh RNA组(空载病毒组)、TRPV4 sh RNA组(TRPV4沉默组)、TRPV4 sh RNA+MPTP组(TRPV4沉默且制备模型组)。开场、悬尾、强迫游泳实验检测行为学变化,免疫印迹及免疫荧光技术检测酪氨酸羟化酶含量变化。结果 TRPV4 sh RNA+MPTP组小鼠与MPTP组相比,行为学评价和酪氨酸羟化酶含量明显提高(P<0.05)。结论 TRPV4沉默能够有效改善帕金森模型小鼠症状及酪氨酸羟化酶含量,表明TRPV4在帕金森病发展中起到重要作用,并有可能成为帕金森病潜在的治疗靶点。
Objective To observe the effect of TRPV4 gene silencing on Parkinson’s disease in mice by lentivirus-mediated sh RNA and to explore the role of TRPV4 in the development of Parkinson’s disease. Methods The lentiviral expression plasmid TRPV4 sh RNA was used to infect substantia nigra pars compacta. After 1 week, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (30 mg · kg -1) Modeling. The mice were randomly divided into 5 groups: normal control group, MPTP group (model group), Scrimble sh RNA group (empty load group), TRPV4 sh RNA group (TRPV4 silencing group), TRPV4 sh RNA + MPTP group Silence and model group was prepared). The changes of tyrosine hydroxylase content in the beginning, tail suspension and forced swimming test were detected by immunoblotting and immunofluorescence. Results Compared with the MPTP group, the TRPV4 sh RNA + MPTP group significantly improved the behavioral assessment and tyrosine hydroxylase (P <0.05). Conclusion TRPV4 silencing can effectively improve the symptoms and tyrosine hydroxylase content in mice with Parkinson’s disease, indicating that TRPV4 plays an important role in the development of Parkinson’s disease and may be a potential therapeutic target for Parkinson’s disease.