A型肉毒毒素轻链胞内持留模型的建立

Establishment of an intracellular persistence model of botulinum toxin type A light chain

摘要:

[目的] 建立A型肉毒毒素轻链胞内持留模型来模拟A型肉毒毒素引起的长期中毒。[方法] 设计构建pcDNA3.1-ALC-GFP重组质粒，提取质粒进行PCR验证，并将其转染进Nureo-2a细胞中表达，利用Western Blot与细胞免疫荧光分析验证ALC-GFP的表达情况及其在细胞内的长时间持留，建立A型肉毒毒素轻链的胞内持留模型，并将该模型用于抗肉毒药物的筛选。[结果] 成功构建pcDNA3.1-ALC-GFP重组质粒并将其转染进Nureo-2a细胞中，验证了ALC-GFP在细胞内具有长时间持留性，成功建立了A型肉毒毒素轻链胞内持留模型，并利用该模型筛选出潜在抗肉毒药物CB3。[结论] 成功建立了A型肉毒毒素轻链胞内持留模型，并初步应用于CS1，CE2和CB3药物筛选，为A型肉毒毒素长期中毒的解毒研究奠定了基础。

Abstract:

[Objective] An intracellular persistence model of light chain of botulinum toxin type A was established to simulate long-term poisoning caused by botulinum toxin type A. [Methods] The recombinant plasmid pcDNA3.1-ALC-GFP was designed and constructed, the plasmid was extracted for PCR verification, and transfected into Nureo-2a cells for expression. Western Blot and cell immunofluorescence analysis were used to verify the expression of ALC-GFP and its long-term persistence in cells. An intracellular persistence model of light chain of botulinum toxin type A was established, and the model was used for the screening of anti-botulinum toxin drugs. [Results] The recombinant plasmid pCDNA3.1-ALC-GFP was successfully constructed and transfected into Nureo-2a cells, which verified the long-term persistence of ALC-GFP in cells. The intracellular persistence model of light chain of botulinum toxin type A was successfully established, and the potential anti-botulinum drug CB3 of botulinum toxin type A was screened by this model. [Conclusions] The intracellular persistence model of light chain of botulinum toxin type A was successfully established and applied to the screening of CS1, CE2 and CB3 drugs, which laid a foundation for the study of detoxification of botulinum toxin A long-term poisoning.