TRPV4沉默对帕金森模型小鼠的影响

Influences of TRPV4 silence on Parkinson's disease model mouse

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。

This study designed to study the influence of TRPV4 silence by lentivirus-mediated RNA interference on Parkinson＇ s disease model mouse, and to investigate the roles of TRPV4 in Parkinson＇ s disease development. Lentivirus expression vector TRPV4 shRNA was used to infect mouse substantia nigra, and a week later, the mice were intraperitoneally injected with 30 mg/kg MPTP to construct Parkinson＇ s disease model. Five experimental groups were set up, including control group, MPTP group, Scrimble shRNA group, TRPV4 shRNA group, and TRPV4 shRNA ＋ MPTP group. The behavioral changes in the five groups of mice were detected by open-field test, tail suspension test and forced swimming test and the tyrosine hydroxylase （TH） levels in all mice were examined by Western blotting and immunofluorescence. TRPV4 shRNA ＋ MPTP group showed significantly better performance in behavioral tests as compared with MPTP group （P〈 0.05）. Also, the TH level was significantly higher in TRPV4 shRNA ＋ MPTP group than that of MPTP group （P〈 0.05）. Taken together, TRPV4 silence can effectively alleviate the symptom of Parkinson＇ s disease model mouse, suggesting that TRPV4 may play an important role in Parkinson＇ s disease development, and may constitute a potential therapeutic target in this disease.