The Biological Study Of Martentoxin, A Peptide Purified From The Venom Of The Scorpion Buthus Martensii Karsch

Part one: The isolation and purification of Martentoxin form the Buthus martensii KarschThe crude toxin form the scorpion (Buthus martensii Karsch) was obtained by Electrical stimulation. A peptide (the purity>95%) was purified form crude toxin by using the ultrafiltration, ion exchange chromatography and reverse phase chromatography. The molecular weight of the peptide was determined by MALDI-TOF-TOF-MS mass spectrometry, and it was 4059 Da. The sequence containing 37 amino acid residues was determined by edman degradation and it was FGLID VKCFA SSECW TACKK VTGSG QGKCQ NNQCR CY. Using the BLAST tools come with the known sequence of scorpion venom Martentoxin 100% consistency in the swiss-prot database.Part two: The impact of Martentoxin on Human umbilical vein endothelial cells releasing nitric oxide induced by TNF-αDespite the Martentoxin found for many years, the relevant biological functions have not been researched very completely. In this project, we used type II collagenase to acute isolation human umbilical vein endothelial cells (Human umbilical vein endothelial cells, HUVECs), and established the inflammatory reaction model by tumor necrosis Factor-α(TNF-α). We investigated the impact of Martentoxin on nitric oxide (NO) release conditions, inducible nitric oxide synthase (iNOS) activity, endothelial nitric oxide synthase (eNOS) mRNA expression, calcium homeostasis, and the apoptosis of glial cells,the results show that Martentoxin inhibited the NO production, iNOS activity and protected the eNOS mRNA.