临床输血与检验
臨床輸血與檢驗
림상수혈여검험
JOURNAL OF CLINICAL TRANSFUSION AND LABORATORY MEDICINE
2009年
3期
199-203
,共5页
红细胞代用品%抗氧化稳定性
紅細胞代用品%抗氧化穩定性
홍세포대용품%항양화은정성
Red blood cell substitutes Antioxidant stability
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70 g/L,MetHb(%) 6.61±1.42,粒径28 nm;制品在4℃、24℃不同保存时间均呈自氧化稳定,37℃、8 h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48 h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.
目的 製備具有抗氧化保護功能的多聚血紅蛋白作為紅細胞代用品,併對其自氧化穩定性和部分生物學特性進行研究.方法 採用戊二醛交聯法製備抗氧化酶交聯的多聚血紅蛋白;Drabkins法測定不同溫度、時間保存下製品MetHb(%)的變化;流式細胞術檢測製品與中性粒細胞孵育後細胞吞噬功能的變化以及製品對T細胞和血小闆的活化作用;凝集試驗檢測血漿中凝血因子活性;Western blot法檢測製品輸註後在小鼠體內的清除速率;ELISA法檢測製品在小鼠體內的抗原性.結果 製備的抗氧化酶交聯的多聚血紅蛋白溶液Hb濃度70 g/L,MetHb(%) 6.61±1.42,粒徑28 nm;製品在4℃、24℃不同保存時間均呈自氧化穩定,37℃、8 h顯示抗氧化酶交聯的多聚血紅蛋白較多聚血紅蛋白有更彊的自氧化穩定性;製品與中性粒細胞孵育後併未影響其吞噬功能,對T細胞和血小闆無活化作用;對凝血因子活性也無明顯影響;製品輸註後48 h仍有檢齣;抗氧化酶交聯的多聚血紅蛋白PolyHb-SOD-CAT抗原性較弱.結論 抗氧化酶交聯的多聚血紅蛋白是一種潛在的安全有效的紅細胞代用品,併具有較彊的抗氧化穩定性.
목적 제비구유항양화보호공능적다취혈홍단백작위홍세포대용품,병대기자양화은정성화부분생물학특성진행연구.방법 채용무이철교련법제비항양화매교련적다취혈홍단백;Drabkins법측정불동온도、시간보존하제품MetHb(%)적변화;류식세포술검측제품여중성립세포부육후세포탄서공능적변화이급제품대T세포화혈소판적활화작용;응집시험검측혈장중응혈인자활성;Western blot법검측제품수주후재소서체내적청제속솔;ELISA법검측제품재소서체내적항원성.결과 제비적항양화매교련적다취혈홍단백용액Hb농도70 g/L,MetHb(%) 6.61±1.42,립경28 nm;제품재4℃、24℃불동보존시간균정자양화은정,37℃、8 h현시항양화매교련적다취혈홍단백교다취혈홍단백유경강적자양화은정성;제품여중성립세포부육후병미영향기탄서공능,대T세포화혈소판무활화작용;대응혈인자활성야무명현영향;제품수주후48 h잉유검출;항양화매교련적다취혈홍단백PolyHb-SOD-CAT항원성교약.결론 항양화매교련적다취혈홍단백시일충잠재적안전유효적홍세포대용품,병구유교강적항양화은정성.
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.
