本文選題：Agkihpin + 蛇毒類凝血酶�。� 參考：《廣東醫(yī)學(xué)》2017年21期

【摘要】：目的探討重組agkihpin將來作為抗血栓藥物的可能性,為將來把重組agkihpin做成抗血栓藥物提供實驗依據(jù)。方法用肽指紋圖譜驗證重組agkihpin分子;以天然agkihpin、生理鹽水(NS)或肝素為對照,分別用TAME水解、纖維蛋白平板、SDS-PAGE和纖維蛋白原凝集法檢測r-agkihpin-1的精氨酸酯酶、纖溶活性、纖維蛋白原溶解和類凝血酶等生物學(xué)活性;以NS或粗毒為對照,用剪尾法和皮下注射法檢測r-agkihpin-1的出血活性。結(jié)果 r-agkihpin-1中38.7%序列、8個肽段與Gen Bank中agkihpin氨基酸序列是一致的。r-agkihpin-1體外水解TAME的比活性為8.74 U/mg。4~16μg/mL劑量r-agkihpin-1可形成纖維蛋白溶圈面積0.50~1.10cm~2,NS組不形成溶圈,差異有統(tǒng)計學(xué)意義(P0.01);16μg/mL劑量r-agkihpin-1可使90%α鏈和70%γ鏈分解,而NS組中纖維蛋白原不被消化,差異有統(tǒng)計學(xué)意義(P0.01)。4~16μg/mL劑量r-agkihpin-1可形成凝塊體積0.07~0.23 mm~3,高于NS組的0.01 mm~3,差異有統(tǒng)計學(xué)意義(P0.05)。體內(nèi)4~16 mg/kg體重劑量的r-agkihpin-1形成血栓干重28.01~22.46 mg/kg體重,低于NS組的35.92 mg/kg體重,差異有統(tǒng)計學(xué)意義(P0.01)。上述活性均稍低于相同劑量的天然agkihpin,差異有統(tǒng)計學(xué)意義(P0.05)。剪尾法、皮下注射法顯示4~16 mg/kg、4~32 mg/kg體重劑量的r-agkihpin-1在小鼠中出血時間和出血活性分別為1.81~11.32 min和0.00~0.01,顯著低于1 mg/kg體重劑量粗毒的30.00 min(P0.01)和7.85(P0.01)。結(jié)論 r-agkihpin-1就是agkihpin的重組蛋白,r-agkihpin-1將來有望成為一種新的抗血栓藥物。
[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% 偽 chain and 70% 緯 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 鹵0.23 mm / 3). The difference was statistically significant (P 0.05). The dry weight of thrombus was 28.01 鹵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.
【作者單位】： 廣西醫(yī)科大學(xué)臨床醫(yī)學(xué)系;廣西醫(yī)科大學(xué)基礎(chǔ)醫(yī)學(xué)院細(xì)胞生物學(xué)與遺傳學(xué)教研室;廣西醫(yī)科大學(xué)附屬腫瘤醫(yī)院影像診斷中心;
【基金】：廣西自然科學(xué)基金資助項目(編號:2014GXNSFAA118172) 廣西大學(xué)生創(chuàng)新創(chuàng)業(yè)訓(xùn)練項目(編號:02610215041C,02610215041X) 廣西壯族自治區(qū)衛(wèi)生和計劃生育委員會自籌經(jīng)費科研課題(編號:Z2015602)
【分類號】：R96

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