猪脑抗吗啡镇痛肽的分离纯化及其抗血清对吗啡耐受影响的初步研究

猪脑组织提取液经SephadexG-50分子筛层析，S-SepharoseFastFlow阳离子交换柱层析及两次HPLC分离得到一分子量为12000，等电点pI7．1的多肽，并测定了其氨基酸组成和N末端部分序列：N-phe-Lys-Gly-Phe-Pro-Asp-Asp／（Lys）-Lys／（Asp）-Asp-Tyr．给昆明小鼠脑室注射或尾静脉注射该肽均能抑制吗啡引起的镇痛作用，其作用随着注射剂量的增大而增强．用BALB／C小鼠制备了该肽的抗血清，脑室注射此抗血清能明显逆转昆明小鼠对吗啡的耐受．因为这种来自猪脑的具有抗阿片镇痛作用的肽有99个氨基酸，所以简称此肽为AOP-99a（anti-oPioidpeptide）．

Purification of an Anti-opioid Peptide from Pig Brain and Study of the Effect of its Anti-Sera on Morphine Tolerance

By Sephadex G-50 chromatography, S-Sepharose Fast Flow ion exchange and RPHPLC,a peptide with molecular weight of 12 000 and pI 7.1 was purified from pig brain. This peptide can antagonize morphine induced analgesia when injected intracereobroventricularally as well as intravenously,and the effects were parallel to the doses. Its amino acid composition and partial sepuence of N-terminus have been determined: N-Phe-Lys-Gly-Phe-Pro-Asp/(Lys)-Lys/(Asp)-Asp-Tyr.This peptide was named AOP-99a which stands for anti-opioid peptide with 990 amino acid residues.Morphine tolerant mice were obtained by administrating increasing doses of morphine for 16 days.The anti-sera against AOP-99a were prepared and when injected intracereobroventricularally,it can significantly reverse morphine tolerance compared to control sera.