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目的探索优化人a突触核蛋白(a-synclein)DNA疫苗(p VAX1-IL-4/SYN-B)免疫保护帕金森病(Parkinson’sdisease)小鼠的泛素蛋白酶体(ubiquitin proteasome)。方法将雄性C57BL/6J小鼠随机分为PBS模型组、空质粒组和优化核酸疫苗和疫苗加蛋白酶体抑制剂组。给予MPTP制备慢性MPTP小鼠模型。然后分别在双侧小鼠后肢胫骨注射PBS、空质粒p VAX1、p VAX1-IL4/SYN-B和p VAX1-IL4/SYN-B核酸疫苗各100μl中,共3次。最后一次注射后,于左侧前脑束(i MFB)区,立体注入乳胞素(lactacystin)。3周后观察动物行为学,免疫组化和Western blot测酪氨酸羟化酶tyrosine hydroxylase TH),同时Western blot检测泛素的表达。结果行为学发现核酸疫苗组较PBS组、空质粒组和疫苗乳胞素组运动症状明显改善,具有统计学意义,TH明显增多,泛素明显减少,具有统计学意义。结论优化a-synclein DNA疫苗对PD的蛋白酶体具有较好的保护作用。
OBJECTIVE: To investigate the ubiquitin proteasome in Parkinson’sdisease mice immunized with a-synclein DNA vaccine (p VAX1-IL-4 / SYN-B). Methods Male C57BL / 6J mice were randomly divided into PBS model group, empty plasmid group and optimized nucleic acid vaccine and vaccine plus proteasome inhibitor group. MPTP was given to prepare a chronic MPTP mouse model. Then PBS was injected into the tibia of the bilateral hindlimbs, and the empty plasmid pVAX1, p VAX1-IL4 / SYN-B and p VAX1-IL4 / SYN-B nucleic acid vaccine in 100μl each for 3 times. After the last injection, lactacystin is stereobjectively injected into the i MFB area. Animal behavior was observed after 3 weeks, tyrosine hydroxylase TH was detected by immunohistochemistry and Western blot, and the expression of ubiquitin was detected by Western blot. Results The behavioral findings showed that compared with the PBS group, the blank plasmid group and the vaccine lactacystatin group, the locomotor activity was significantly improved in the nucleic acid vaccine group, which was statistically significant. TH was significantly increased and ubiquitin was significantly decreased, with statistical significance. Conclusion The optimized a-synclein DNA vaccine has a good protective effect on PD proteasome.