摘要
目的制备具有抗氧化保护功能的多聚血红蛋白作为红细胞代用品,并对其自氧化稳定性和部分生物学特性进行研究。方法采用戊二醛交联法制备抗氧化酶交联的多聚血红蛋白;Drabkins法测定不同温度、时间保存下制品MetHb(%)的变化;流式细胞术检测制品与中性粒细胞孵育后细胞吞噬功能的变化以及制品对T细胞和血小板的活化作用;凝集试验检测血浆中凝血因子活性;Western blot法检测制品输注后在小鼠体内的清除速率;ELISA法检测制品在小鼠体内的抗原性。结果制备的抗氧化酶交联的多聚血红蛋白溶液Hb浓度70g/L,MetHb(%)6.61±1.42,粒径28nm;制品在4℃、24℃不同保存时间均呈自氧化稳定,37℃、8h显示抗氧化酶交联的多聚血红蛋白较多聚血红蛋白有更强的自氧化稳定性;制品与中性粒细胞孵育后并未影响其吞噬功能,对T细胞和血小板无活化作用;对凝血因子活性也无明显影响;制品输注后48h仍有检出;抗氧化酶交联的多聚血红蛋白PolyHb-SOD-CAT抗原性较弱。结论抗氧化酶交联的多聚血红蛋白是一种潜在的安全有效的红细胞代用品,并具有较强的抗氧化稳定性。
Objective To produce antioxidant enzymes crosslinked polyhemoglobin as red blood cell substitutes,and to study its auto-oxidative stability and partial biological characters. Methods Glutaraldehyde was used to crosslink hemogloibin and antioxidant enzymes. Drabkins method was used to measure the change of MetHb（%） of the product preserved in different time and temperature. The change of neutrophil phagocytosis of the cells,activation of T lymphocytes and platelet incubated with products was estimated using flow cytometry. The aggregation test was made to measure the function of serum accelerin incubated with products. The rate of elimination of the products infused in the mouse was detected using western blot. Results Characters of the product were as follows ：Hb 70 g/L,MetHb （%） 6.61±1.42, particle size 28 nm; products preserved in different time and temperature showed autoxidative stablity. And the polyhemoglobin crosslinked with antioxidant enzymes appeared to be more stable than polyhemoglobin during the incubation（37℃ 8 h）. No significant influence on the function of neutrophil phagocytosis and serum accelerin aggregation was measured. T lymphocytes and platelet stayed inactivated during the incubation. The product could still be detected 48 h after infused into the mouse. ELISA tests showed weak or no antigenicity. Conclusion The polyhemoglobin crosslinked with antioxidant enzymes can be used as a kind of potential safe and effective red blood cell substitute with better antioxidant stability.
出处
《临床输血与检验》
CAS
2009年第3期199-203,共5页
Journal of Clinical Transfusion and Laboratory Medicine
基金
上海市科委基金(No.0652nm027)资助
关键词
红细胞代用品
抗氧化稳定性
Red blood cell substitutes Antioxidant stability
