A Kunming mice local skin necrosis model of the cytotoxin of Chinese cobra
Objective To establish a Kunming mice's local skin necrosis model by intradermal injection of Chinese cobra's cytotoxin(CTX).Methods Cobra cytotoxin(CTX-X)was isolated from the crude venom of Chinese cobra by gel chromatography + ion chromatography.30 mice were randomly divided into 10 groups:Normal saline group,10 μg crude venom group,20 μg crude venom group,25 μg crude venom group,10 μg CTX-X group,20 μg CTX-X group,30 μg CTX-X group,40 μg CTX-X group,50 μg CTX-X group,60 μg CTX-X group,and 3 rats in each group.Observe the diameter of skin necrosis and pathological changes at the injection site for 72 h.Results(1)The crude cobra venom was separated by Sephadex G-50 gel chromatography to obtain two protein peaks(F1,F2),and the second peak(F2)where the CTX-X was located was separated by CM SepharoseTMCL-6B cation-exchange column chro-matography to obtain four peaks,including F1,F2,F3,and F4.CTX-X activity was determined by in vivo mouse assay and CTX-X purity was determined by SDS-PAGE.(2)The diameter of intradermal necrosis in the CTX-X group of mice increased with increasing CTX-X dose,and the mean intradermal necrosis diameter in the 51.21 μg CTX-X group reached 5mm;In the crude group,the mortality rate of mice within 72 h increased with the increase of the intradermal injection dose of crude poison.When the intradermal injection dose of crude poison was<25 μg,mice were executed and dissected after 72 h of injection,no skin necrosis was observed at the injection site,and when the intradermal injection dose of crude poison was≥25 μg,all the mice were dead within 3 h of injection,and no obvious necrosis was observed at the intradermal injection site.Conclusion A mice model of CTX-X-induced local skin necrosis in Chinese cobras was successfully constructed by intradermal injection of CTX-X obtained by Sephadex G-50 gel filtration +CM SepharoseTMCL-6B cation exchange,which was highly stable and reproducible.
Chinese cobra's venomCytotoxinLocal skin necrosisAnimal model
NETL
NSTL
万方数据
维普
