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目的探讨维生素C(VC)和氮-乙酰半胱氨酸(NAC)对悬浮红细胞保存过程中氧化应激损伤的抑制作用,二者介入悬浮红细胞的量效关系。方法取新鲜悬浮红细胞(采集制备至实验开始3 d)8袋(2 U/袋),使每袋均分为9份(40 m L/份),用无菌方式转移至50 m L塑料血袋,设VC(实验组):取4份分别加入0.1、1、10和100mmol/L的VC溶液各约4.44 m L,使其终浓度为0.01、0.1、1和10 mmol/L;NAC(实验组):取4份分别加入0.1、1、10和100 mmol/L的NAC溶液各约4.44 m L,使其终浓度为0.01、0.1、1和10 mmol/L;对照组:剩余1份加入等体积的等渗PBS。3组红细胞置于(4±2)℃保存,分别于保存7、14、21、28及35 d取样(6 m L/次)测定红细胞变形性、渗透脆性(g/L)、ATP、谷胱甘肽过氧化物酶(GSH-Px)。结果 RBC保存初期,与对照组相比,1和10 mmol/L VC实验组对200-1、1 000-1剪切率EI、渗透脆性(g/L)、胞内ATP(μmol/g Hb)、GSH-Px(U/L)这4个指标均有改善:0.290±0.022 vs 0.297±0.018 vs 0.282±0.026、0.527±0.029 vs 0.538±0.023vs 0.520±0.043、4.892±0.128 vs 4.908±0.741 vs 5.000±0.068、4.82±0.49 vs4.74±0.52 vs 4.45±0.57、89.24±5.76vs 90.01±2.20 vs 75.51±16.03(P<0.05);1和10 mmol/L NAC实验组在200-1、1 000-1剪切率EI值、渗透脆性、胞内ATP(μmol/g Hb)、GSH-Px(U/L)分别为:0.284±0.023 vs 0.298±0.012 vs 0.282±0.026、0.540±0.024 vs 0.550±0.027 vs 0.520±0.043、0.511±0.56vs 5.000±0.068 vs 4.45±0.57、90.59±9.9 vs 99.06±27.8 vs75.51±16.03,其他浓度实验组红细胞没有明显的改变或规律性;保存末期,与对照组相比,1 mmol/L VC和NAC实验组对渗透脆性(g/L)及GSH-Px(U/L)这2个指标有改善:0.260±0.017vs 0.257±0.027vs 0.241±0.024、0.507±0.019 vs 0.491±0.027 vs 0.485±0.036、87.89±10.83 vs 86.12±8.71 vs 77.75±8.17(P<0.05)。结论适宜浓度的VC和NAC能够抑制悬浮红细胞保存过程中的氧化应激损伤,改善悬浮红细胞的保存质量。
Objective To investigate the inhibitory effect of vitamin C (VC) and nitrogen-acetylcysteine (NAC) on the oxidative stress injury in suspension of erythrocytes, and to investigate the dose-response relationship between the two interventional erythrocytes. Methods Freshly suspended erythrocytes were collected (8 h (2 U / bag) from the collection to the first 3 days of the experiment), and each bag was divided into 9 portions (40 m L / portion) and sterilely transferred to 50 ml plastic blood Bags, set VC (experimental group): Take 4 copies of VC solution of 0.1, 1, 10 and 100 mmol / L, respectively, about 4.44 m L to make final concentrations of 0.01, 0.1, 1 and 10 mmol / Experimental group): Take 4 portions of about 4.44 m L of NAC solution of 0.1, 1, 10 and 100 mmol / L, respectively, to make the final concentration of 0.01, 0.1, 1 and 10 mmol / L; the control group: the remaining 1 Equal volume of isotonic PBS was added. The erythrocytes in 3 groups were stored at (4 ± 2) ℃, and the erythrocyte deformability, osmotic fragility (g / L), ATP and valley were determined at 6, 14, 21, 28 and 35 d after storage Glutathione peroxidase (GSH-Px). Results In the early stage of RBC preservation, compared with the control group, the shear stress of 200-1,1000-1 EI, osmotic fragility (g / L), intracellular ATP (μmol / g Hb ), GSH-Px (U / L) all improved: 0.290 ± 0.022 vs 0.297 ± 0.018 vs 0.282 ± 0.026,0.527 ± 0.029 vs 0.538 ± 0.023 vs 0.520 ± 0.043, 4.892 ± 0.128 vs 4.908 ± 0.741 vs 5.000 ± 0.068,4.82 ± 0.49 vs4.74 ± 0.52 vs 4.45 ± 0.57,89.24 ± 5.76 vs 90.01 ± 2.20 vs 75.51 ± 16.03 (P <0.05). In the NAC group of 1 and 10 mmol / L, -1, shear rate EI, osmotic fragility, intracellular ATP (μmol / g Hb) and GSH-Px (U / L) were 0.284 ± 0.023 vs 0.298 ± 0.012 vs 0.282 ± 0.026 and 0.540 ± 0.024 vs 0.550 ± 0.027 vs 0.520 ± 0.043,0.511 ± 0.56vs 5.000 ± 0.068 vs 4.45 ± 0.57,90.59 ± 9.9 vs 99.06 ± 27.8 vs75.51 ± 16.03, no significant change or regularity of erythrocytes in other concentrations of experimental group; at the end of preservation, compared with the control group Compared with the 1 mmol / L VC and NAC groups, the two indexes of osmotic fragility (g / L) and GSH-Px (U / L) were improved: 0.260 ± 0.017 vs 0.257 ± 0.027 vs 0.241 ± 0.024 and 0.507 ± 0.019 vs 0.491 ± 0.027 vs 0.485 ± 0.036, 87.89 ± 10.83 vs 86.1 2 ± 8.71 vs 77.75 ± 8.17 (P <0.05). Conclusion Appropriate concentration of VC and NAC can inhibit the oxidative stress injury in the process of suspension of erythrocytes and improve the quality of the preserved erythrocytes.