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目的研究四环素对朊病毒羊瘙痒因子263K(PrPSc)体外蛋白酶抗性和实验动物感染性的影响。方法以不同浓度的四环素与PrPSc进行不同时间的作用,经蛋白酶K的处理后以Westernblot方法检测具有蛋白酶抗性的PrPSc蛋白;将经四环素处理的PrPSc样品分组颅内接种金黄地鼠,观察动物发病,以Westernblot方法检测发病动物脑组织中PrPSc的存在情况。结果四环素与PrPSc的体外作用可明显地降低或消除PrPSc对蛋白酶K消化的抵抗能力,并呈明显的剂量效应相关性。与对照组相比[平均潜伏期(53.75±0.50)d],经5mmol/L和20mmol/L四环素处理的高滴度PrPSc样品的实验动物感染潜伏期延长,潜伏期分别为(61.50±1.73)d(P<0.01)和(59.50±0.58)d(P<0.05),差异有统计学意义。结论四环素能够有效降低PrPSc的体外蛋白酶抗性,并延长PrPSc感染动物发病潜伏期。
Objective To investigate the effect of tetracycline on in vitro protease resistance and experimental animal infectivity of prion pruritus factor 263K (PrPSc). Methods Different concentrations of tetracycline and PrPSc were used for different time periods. Proteinase K-resistant PrPSc protein was detected by Western blot after treated with proteinase K, and tetracycline-treated PrPSc samples were intracranially inoculated with golden hamster Western blot was used to detect the presence of PrPSc in the brain tissue of the diseased animals. RESULTS: The in vitro effect of tetracycline and PrPSc significantly reduced or eliminated the resistance of PrPSc to protease K digestion and showed a significant dose-response relationship. Compared with the control group (mean latency (53.75 ± 0.50) d], the infection latency of experimental animals with high titers of PrPSc treated with 5mmol / L and 20mmol / L tetracycline was prolonged with latency of (61.50 ± 1.73) d <0.01) and (59.50 ± 0.58) d (P <0.05), the difference was statistically significant. Conclusion Tetracycline can effectively reduce protease resistance of PrPSc in vitro and prolong the incubation period of PrPSc-infected animals.