Keywords

Bat3, Prp106-126 Toxic Peptide, Neuroapoptosis, Transmissible Spongiform Encephalopathy, Primary Neurons

Abstract

To detect the effect of bat3 protein overexpression on neuronal apoptosis caused by prp106-126 toxic peptide. First, the biological toxicity of prp106-126-fitc toxic peptide with FITC tag was detected through immunofluorescence, cck8 cell activity detection kit and western blotting. Primary neurons and neuro2a cells were treated with prp106-126-fitc and prp106-126 neurotoxic peptides respectively, and the changes in bat3 expression in the stimulated primary neurons and neuro2a cells were detected by immunofluorescence technology and western blotting methods. . The constructed pcdna-3.1-ha-bat3 plasmid was transfected into primary neurons by lipofection method, and cell activity was detected with cck8; in the same way, the plasmid was transfected into the mouse neuroma cell line neuro2a. Use tunel apoptosis detection kit to analyze the apoptosis level of n2a cells transfected with bat3 or control group; westernblot detects the changes of cytochrome c and bcl-2 in the cytoplasm before and after treatment with prp106-126 neurotoxic peptide to further understand bat3 Mechanisms that play a role in neuronal apoptosis. The results showed that prp106-126-fitc and prp106-126 neurotoxic peptides have similar neurotoxicity. After being stimulated by peptides, bat3 undergoes nuclear export in both primary neurons and the passaged cell line neuro2a, and bat3 is transferred from the nucleus into the cytoplasm. Overexpression of bat3 increased the cell activity of neurons stimulated by the prp106-126 toxic peptide, alleviated the apoptosis of neuro2a, inhibited the excessive release of cytochrome c caused by the prp106-126 toxic peptide, and maintained cell Stabilization of the internal anti-apoptotic protein bcl-2. The expression of bat3 protein alleviates the apoptosis phenomenon of nerve cells caused by prp106-126 toxic polypeptide, which provides a basis for further elucidating the therapeutic mechanism of this protein on neuron loss caused by prions.

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Original research done by Song Zhiqi, Zhao Wenjie, Yang Lifeng, Wang Yunsheng, Zhu Ting, Cheng Guangyu, Zhou Xiangmei, Zhao Deming

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