Isolation and purification of α-neurotoxin and fluorescence imaging analysis of intestinal tract after rectum administration in mice
Objective To isolate and purify the short-chain α-neurotoxin active ingredient from the crude venom of cobra,and to observe the distribution and dynamic process of fluorescent dye-labeled α-neurotoxin in the intestinal tract after rectal administration to mice.Methods Electrophoretically pure α-neurotoxin was isolated and purified using CM Sepharose Fast Flow cation exchange chromatography and Sephadex G-50 gel filtration chromatography.Four C57 mice were divided into two groups:control and experimen-tal.In the experimental group,the fluorescent dye CY7-SE was conjugated with α-neurotoxin to each other.The fluorescence intensity distribution of CY7-SE-labeled α-neurotoxin in the intestine was detected by using an imager after 1h and 3h of rectal administration,respectively.Results Electrophoretically pure α-neurotoxin was obtained by isolation and purification.α-neurotoxin administered rectally for 1h showed stronger fluorescence intensity in the small intestine and stomach,and weaker fluorescence intensity in the cecum and colon than that in the control group.α-neurotoxin administered rectally for 3h showed weaker fluorescence intensity in the small in-testine,cecum,colon,and stomach than that in the control group.Conclusion The α-neurotoxin isolated and purified from the crude venom of cobra was first enriched in the small intestine and stomach of mice,and then absorbed and metabolized by the intestinal muco-sa after 3h.This study provides an experimental basis for the development of a neurotoxin formulation for rectal delivery.
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万方数据
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