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目的观察帕金森小鼠模型中TLR4通路对Beta-catenin信号的表达影响。方法 C57BL小鼠和TLR4敲除(TLR4knock out,TLR4 KO)小鼠各36只,分别注射生理盐水和1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)。采用RT-PCR、Westernblot检测4组之间的Beta-catenin的m RNA和蛋白表达变化,进一步采用免疫荧光的方法检测黑质区星形胶质细胞中Beta-catenin的共表达变化。结果 RT-PCR及Western blot结果显示,与生理盐水注射组小鼠相比,C57BL、TLR4KO小鼠在MPTP注射后,mRNA和蛋白表达明显升高(P<0.01);并且,与C57BL小鼠MPTP注射组相比,TLR4KO小鼠MPTP注射组Beta-catenin的mRNA和蛋白表达明显升高(P<0.01)。进一步的免疫荧光结果显示,与C57BL小鼠MPTP注射组相比,TLR4KO小鼠MPTP注射组星形胶质细胞(GFAP)中表达的beta-catenin明显升高。结论 TLR4通路在亚急性帕金森MPTP模型中,对黑质beta-catenin蛋白的表达起重要调控作用,Toll样信号与Beta-catenin信号这种互动关系可能在帕金森发病机制中起着重要的作用。
Objective To investigate the effect of TLR4 pathway on the expression of Beta-catenin in Parkinson’s mouse model. Methods Thirty-six C57BL mice and TLR4 knockout (TLR4knock out) mice were injected with normal saline and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) . The mRNA and protein expressions of Beta-catenin in four groups were detected by RT-PCR and Western blot. The co-expression of Beta-catenin in substantia nigra astrocytes was detected by immunofluorescence. Results RT-PCR and Western blot showed that the mRNA and protein expressions of C57BL and TLR4KO mice were significantly increased (P <0.01) after MPTP injection compared with saline injection group. Compared with C57BL mice, MPTP Compared with the injection group, the mRNA and protein expression of Beta-catenin in TLR4KO mice MPTP injection group was significantly increased (P <0.01). Further immunofluorescence results showed that the expression of beta-catenin in astrocytes (GFAP) of MPTP injection group was significantly increased in TLR4KO mice compared with MPTP injection group in C57BL mice. Conclusion TLR4 pathway plays an important regulatory role in the substantia nigra beta-catenin protein expression in the subacute Parkinson’s MPTP model. The interaction between Toll-like signal and Beta-catenin signaling may play an important role in the pathogenesis of Parkinson’s disease .