新重组蛇毒类凝血酶r-agkihpin-1的抗血栓活性

发布时间：2018-05-30 10:42

本文选题：Agkihpin + 蛇毒类凝血酶　；参考：《广东医学》2017年21期

【摘要】：目的探讨重组agkihpin将来作为抗血栓药物的可能性,为将来把重组agkihpin做成抗血栓药物提供实验依据。方法用肽指纹图谱验证重组agkihpin分子;以天然agkihpin、生理盐水(NS)或肝素为对照,分别用TAME水解、纤维蛋白平板、SDS-PAGE和纤维蛋白原凝集法检测r-agkihpin-1的精氨酸酯酶、纤溶活性、纤维蛋白原溶解和类凝血酶等生物学活性;以NS或粗毒为对照,用剪尾法和皮下注射法检测r-agkihpin-1的出血活性。结果 r-agkihpin-1中38.7%序列、8个肽段与Gen Bank中agkihpin氨基酸序列是一致的。r-agkihpin-1体外水解TAME的比活性为8.74 U/mg。4~16μg/mL剂量r-agkihpin-1可形成纤维蛋白溶圈面积0.50~1.10cm~2,NS组不形成溶圈,差异有统计学意义(P0.01);16μg/mL剂量r-agkihpin-1可使90%α链和70%γ链分解,而NS组中纤维蛋白原不被消化,差异有统计学意义(P0.01)。4~16μg/mL剂量r-agkihpin-1可形成凝块体积0.07~0.23 mm~3,高于NS组的0.01 mm~3,差异有统计学意义(P0.05)。体内4~16 mg/kg体重剂量的r-agkihpin-1形成血栓干重28.01~22.46 mg/kg体重,低于NS组的35.92 mg/kg体重,差异有统计学意义(P0.01)。上述活性均稍低于相同剂量的天然agkihpin,差异有统计学意义(P0.05)。剪尾法、皮下注射法显示4~16 mg/kg、4~32 mg/kg体重剂量的r-agkihpin-1在小鼠中出血时间和出血活性分别为1.81~11.32 min和0.00~0.01,显著低于1 mg/kg体重剂量粗毒的30.00 min(P0.01)和7.85(P0.01)。结论 r-agkihpin-1就是agkihpin的重组蛋白,r-agkihpin-1将来有望成为一种新的抗血栓药物。
[Abstract]:Objective to explore the possibility of recombinant agkihpin as an antithrombotic in the future and to provide experimental evidence for making recombinant agkihpin into antithrombotic in the future. Methods the recombinant agkihpin molecule was verified by peptide fingerprinting, and the TAME hydrolysis, fibrin plate SDS-PAGE and fibrinogen agglutination method were used to detect the arginine esterase and fibrinolytic activity of r-agkihpin-1, using natural agkihpin (NS) or heparin as control. Fibrinolysis and thromboplastin were used to detect the bleeding activity of r-agkihpin-1 with NS or crude toxin as control. Results 38.7% of the r-agkihpin-1 sequences were identical with the amino acid sequence of agkihpin in Gen Bank. The specific activity of TAME hydrolyzed by r-agkihpin-1 in vitro was 8.74 U/mg.4~16 渭 g/mL (r-agkihpin-1). The fibrinolytic circle area of 0.50 ~ 1.10 cm ~ (-2) N group was not formed in NS group. The difference was statistically significant (P 0.01 渭 g/mL) r-agkihpin-1 could decompose 90% 伪 chain and 70% 纬 chain, but fibrinogen was not digested in NS group. The difference was statistically significant (P 0.01N 路4N 16 渭 g/mL r-agkihpin-1), which was higher than that in NS group (0.07 卤0.23 mm / 3). The difference was statistically significant (P 0.05). The dry weight of thrombus was 28.01 卤22.46 mg/kg, which was lower than that of NS group (35.92 mg/kg), and the difference was statistically significant (P 0.01). The above activities were slightly lower than those of natural agkihpinat the same dose, and the difference was statistically significant (P 0.05). The results showed that the bleeding time and bleeding activity of r-agkihpin-1 at the body weight dose of 4 ~ 16 mg 路kg ~ (-1) 路kg ~ (-1) mg/kg were 1.81 ~ 11.32 min and 0.000. 01 min, respectively, which were significantly lower than those of 30. 00? min P0.01 and 7.85? P0. 01 of 1 mg/kg body weight dose. Conclusion r-agkihpin-1 is a recombinant protein of agkihpin, r-agkihpin-1, which is expected to become a new antithrombotic drug in the future.
【作者单位】：广西医科大学临床医学系;广西医科大学基础医学院细胞生物学与遗传学教研室;广西医科大学附属肿瘤医院影像诊断中心;
【基金】：广西自然科学基金资助项目(编号:2014GXNSFAA118172) 广西大学生创新创业训练项目(编号:02610215041C,02610215041X) 广西壮族自治区卫生和计划生育委员会自筹经费科研课题(编号:Z2015602)
【分类号】：R96

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