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目的探讨不同氧化模型制备时红细胞聚集性和变形性的变化。方法从15只大鼠颈动脉采集血液9m L/只,离心去除白细胞,获得红细胞重悬液分为4组(n=3):空白对照组(不加入任何氧化剂);过氧化氢(H2O2)组(终浓度为0.5、5、8 mmol/L);吩嗪硫酸甲酯(PMS)组(终浓度为25、50、100μmol/L),亚硝酸钠(Na NO2)组(终浓度为0.5、1、1.5 mmol/L);37℃水浴1 h,自体血浆调节红细胞压积至40%,测定红细胞聚集性和变形性。结果红细胞聚集指数:H2O2组呈下降趋势,终浓度8 mmol/L时的2.5±1.3,较对照组的45.3±1.5下降明显(P<0.05);PMS和Na NO2组则较对照组均无明显变化(P>0.05)。红细胞聚集时间(s):与对照组的0.89±0.13相比,H2O2组作用浓度为5、8 mmol/L时明显升高,分别为1.68±0.2、1.85±0.14(P<0.05);PMS组各作用浓度未见明显影响(P>0.05);Na NO2组各终浓度作用后,虽然分别降低至0.54±0.06、0.62±0.03和0.78±0.08,但也显示作用浓度越高降低越慢。红细胞变形性:各个剪切速率(SHR)下,对照组EI分别为28.1±0.45、40.6±0.92、42.4±1.09、43.2±1.12、44±1.21。3种浓度H2O2作用后,不同SHR下EI均明显下降(SHR=100 s-1时EI分别为18.8±3.78、11.3±2.36、6.9±4.89,P<0.05);3种浓度PMS作用后,不同SHR下EI均明显下降(SHR=100 s-1时EI分别为16.8±2.27、8.5±0.64、5.6±0.07,P<0.05);3种浓度Na NO2作用后,SHR=100 s-1时,变形性降低(分别为23.9±1.41、20.8±0.01、21.9±041,P<0.05),但随着SHR的增大,变形性有升高趋势,SHR≥600 s-1时,Na NO2组红细胞变形性明显高于对照组(SHR=1000 s-1时EI分别为48.3±0.73、49.2±0.1、48.3±0.65,P<0.05)。结论不同氧化剂对红细胞聚集性和变形性影响不同,提示在体外建立红细胞氧化损伤模型时应根据实验目的选取符合要求的氧化剂。
Objective To investigate the changes of erythrocyte aggregation and deformability during the preparation of different oxidation models. Methods Blood was collected from 15 carotid arteries in 15 rats and centrifuged to remove leukocytes. The erythrocyte suspension was divided into 4 groups (n = 3): blank control group (without any oxidant); hydrogen peroxide (H2O2) (Final concentration of 0.5, 5, 8 mmol / L), PMS group (final concentration of 25, 50 and 100 μmol / L), sodium nitrite group , 1, 1.5 mmol / L). After 37 ℃ water bath for 1 h, autologous plasma was used to adjust the hematocrit to 40%. The erythrocyte aggregation and deformability were measured. Results The index of erythrocyte aggregation decreased in H2O2 group, but decreased significantly in 2.5 mmol / L and 8 mmol / L group compared with 45.3 ± 1.5 in control group (P <0.05), but no significant difference between PMS group and NaNO2 group Change (P> 0.05). The erythrocyte aggregation time (s): Compared with the control group (0.89 ± 0.13), the concentration of H2O2 group increased significantly at 5 and 8 mmol / L, which were 1.68 ± 0.2 and 1.85 ± 0.14 respectively (P <0.05) (P> 0.05). After the final concentrations of Na NO2 group were decreased to 0.54 ± 0.06,0.62 ± 0.03 and 0.78 ± 0.08 respectively, they also showed that the higher concentration and the lower the concentration, the lower the concentration. Erythrocyte deformability: At each shear rate (SHR), the EI of the control group were 28.1 ± 0.45, 40.6 ± 0.92, 42.4 ± 1.09, 43.2 ± 1.12 and 44 ± 1.21, respectively. (SHR = 100 s-1, EI 18.8 ± 3.78,11.3 ± 2.36,6.9 ± 4.89, respectively, P <0.05). After three concentrations of PMS, 1 at 16.8 ± 2.27, 8.5 ± 0.64 and 5.6 ± 0.07 respectively, P <0.05). Decreased deformability was observed at SHR = 100 s-1 after treated with NaNO 2 (23.9 ± 1.41 and 20.8 ± 0.01, 21.9 ± 041, P <0.05). However, with the increase of SHR, the deformability tended to increase. When SHR≥600 s-1, the erythrocyte deformability of NaNO2 group was significantly higher than that of the control group (SHR = 1000 s EI were 48.3 ± 0.73, 49.2 ± 0.1, 48.3 ± 0.65 respectively, P <0.05). Conclusion Different oxidants have different effects on erythrocyte aggregation and deformability, suggesting that oxidant should be selected according to the purpose of experiment when establishing erythrocyte oxidative damage model in vitro.