不同浓度葛根素对G6PD缺乏症患者红细胞的溶血率及H2O2耐受力影响

Effects of different concentrations of perarin injection on hemolysis rate and H2O2 tolerancein patients with glucose-6-phosphate dehydrogenase deficiency

目的观察不同浓度葛根素对葡萄糖-6-磷酸脱氢酶(G6PD)缺乏症患者红细胞的溶血率及H2O2耐受力影响。方法选择15例G6PD缺乏症患者为观察组,10例健康体检者为对照组,取两组外周血中的红细胞;在观察组和对照组红细胞悬液中,分别加入浓度为1. 15、1. 35、1. 55、1. 75、1. 95、2. 15、2. 35、2. 55、2. 75、2. 95、3. 15、3. 35 mg/m L的葛根素注射液,37℃孵育1 h后检测红细胞溶血率。取观察组红细胞悬液分别加入终浓度为1. 15mg/m L的葛根素注射液(葛根素组),0. 9%氯化钠注射液(NS组),再分别加入浓度为8. 1、9. 0、9. 9、10. 8、11. 7、12. 6、13. 5 mg/m L的H2O2溶液,孵育后测溶血率;以8. 1 mg/m L的H2O2与观察组红细胞混合同时分别加入浓度为0. 60、0. 75、0. 90、1. 05、1. 20、1. 35、1. 50 mg/m L的葛根素注射液,孵育1 h后测定溶血率。结果①观察组葛根素开始溶血浓度(1. 48±0. 18) mg/m L,低于对照组(1. 69±0. 21) mg/m L(P <0. 05)。加入上述浓度的葛根素注射液,观察组的平均溶血率分别为2. 89%、4. 74%、7. 51%、13. 62%、29. 67%、49. 66%、73. 47%、85. 70%、89. 92%、95. 11%、96. 40%、96. 88%,对照组分别为1. 32%、2. 12%、5. 17%、5. 92%、15. 97%、21. 25%、27. 75%、38. 70%、57. 75%、71. 32%、81. 97%、88. 98%,从加入的葛根素注射液浓度为1. 75 mg/m L开始,随着葛根素注射液浓度的增高,同一葛根素注射液浓度下观察组溶血率高于对照组(P均<0. 05)。②葛根素组加入上述浓度的H2O2溶液后溶血率分别为9. 89%、12. 49%、17. 36%、25. 03%、31. 62%、42. 08%、47. 48%,NS组分别为10. 43%、12. 52%、20. 73%、31. 70%、44. 76%、53. 12%、65. 33%,从H2O2浓度为9. 9 mg/m L开始,随着H2O2浓度递增,葛根素组及NS组的溶血率均上升;在同一H2O2浓度下,葛根素组的溶血率均低于NS组(P均<0. 05)。以8. 1 mg/m L的H2O2与观察组红细胞混合后,同时加入浓度依次为0. 6、0. 75、0. 90、1. 05、1. 20、1. 35、1. 50 mg/m L的葛根素注射液后溶血率分别为6. 11%、5. 95%、7. 28%、11. 81%、18. 91%、29. 48%、40. 59%;葛根素注射液浓度在0. 6~0. 9mg/m L之间的溶血率比较,P均> 0. 05;葛根素注射液浓度1. 05~1. 50 mg/m L范围内,G6PD缺乏症患者红细胞溶血率反而随葛根素注射液浓度的增高而增高(P均<0. 05)。结论与健康体检者的红细胞相比,1. 75~3. 35mg/m L的葛根素注射液增加G6PD缺乏症患者红细胞溶血率,即G6PD缺乏症患者红细胞对1. 75-3. 35 mg/m L的葛根素注射液的耐受力减低。H2O2浓度为9. 9~13. 5 mg/m L时,随着H2O2浓度递增,1. 15 mg/m L葛根素可使G6PD缺乏症患者红细胞溶血率降低。低浓度(0. 60~0. 90 mg/m L)葛根素注射液能提高G6PD缺乏症患者红细胞对8. 1 mg/m L H2O2的耐受能力,但葛根素注射液浓度为1. 05~1. 50 mg/m L时,G6PD缺乏症患者红细胞对8. 1mg/m L H2O2的耐受能力降低。

Objective To observe the effects of different concentrations of puerarin injection on hemolysis rate and H2O2 tolerancein patients with glucose-6-phosphate dehydrogenase(G6PD)deficiency.Methods Fifteen G6PD deficiency patients(observation group)and 10 healthy subjects(control group)were selected.The red blood cells(RBCs)were collected from the two groups.The puerarin injections of 1.15,1.35,1.55,1.75,1.95,2.15,2.35,2.55,2.75,2.95,3.15,and 3.35 mg/m L were added to the suspension of RBCs of two groups,respectively.We detected the hemolysis rate after incubation in 37℃water for 1 hour.In the observation group,we added puerarin injection with a final concentration of1.15 mg/m L(puerarin group)and 0.9%sodium chloride injection(NS group)to the red blood cell suspension,followed by 8.1,9.0,and 9.9,10.8,11.7,12.6,and 13.5 mg/m L H2O2 solution,and then we measured hemolysis rate after incubation.In the control group,the G6PD deficiency RBCs were treated with 8.1 mg/m L H2O2,followed by 0.60,0.75,0.90,1.05,1.20,1.35,and 1.50 mg/m L puerarin,and then we detected the hemolysis rate after incubation for 1 hour.Results①Thehemolysisbegan at the concentration of(1.48±0.18)mg/m L in the observation group,which was lower than that of the control group[(1.69±0.21)mg/m L],with statistically significant different(P<0.05).After we added the above concentrations of puerarin injections,the average hemolysis rates of the observation group were 2.89%,4.74%,7.51%,13.62%,29.67%,49.66%,73.47%,85.70%,89.92%,95.11%,96.40%,and 96.88%,respectively,and were 1.32%,2.12%,5.17%,5.92%,15.97%,21.25%,27.75%,38.70%,57.75%,71.32%,81.97%,and 88.98%,respectively,in the control group.Starting from the addition of puerarin injection at 1.75 mg/m L,with the increasing concentrations of pueraria injections,the hemolysis rates of the observation group were significantly higher than those of the control group at the same concentration of pueraria injection(all P<0.05).②During the increasing H2O2 addition,the average hemolysis rates of the pueraringroup were 9.89%,12.49%,17.36%,25.03%,31.62%,42.08%,and 47.48%,respectively,and were 10.43%,12.52%,20.73%,31.70%,44.76%,53.12%,and 65.33%,respectively,in the NS group.Starting from the addition of H2O2 at 9.9 mg/m L,with the increasing concentrations of H2O2,both the hemolysis rates of pueraria group and NS group increased.At the same concentration of H2O2,the hemolysis rate of puerarin group was significantly lower than that of NS group(P<0.05).In the observation group,the RBCs were treated with 8.1 mg/m L H2O2 and 0.60,0.75,0.90,1.05,1.20,1.35,and 1.50 mg/m L puerarin,and then the hemolysis rates were 6.11%,5.95%,7.28%,11.81%,18.91%,29.48%,and 40.59%,respectively.There was no significant difference in the hemolysis rate between the 0.6-0.9 mg/m L puerarin(P>0.05).Within the range of 1.05-1.50 mg/m L of puerarin,the hemolysis rate of G6PD deficient RBCs increased with the increase of puerarin concentration(P<0.05).Conclusion Within the range of 1.75-3.35 mg/m L of puerarin,erythrocyte hemolysis rate in patients with G6PD deficiency increases compared with that of the healthy subjects,which means the tolerance of erythrocyte to1.75-3.35 mg/m L puerarin in G6PD deficiency is decreasing.In the range of 9.9-13.5 mg/m L of H2O2,as the concentration increases,1.15 mg/m L pueraria could reduce erythrocyte hemolysis rate in patients with G6PD deficiency.Low concentration(0.60-0.90 mg/m L)of pueraria injection could improve the tolerance of G6PD deficient RBCs to 8.1 mg/m L H2O2,but when the concentration of pueraria injection reaches 1.05-1.50 mg/m L,the tolerance of G6PD deficient RBCs to 8.1 mg/m L H2O2 decreases.