【作者】 刘晓华；

【导师】 刘成玉； 谭齐贤；

【作者基本信息】 青岛大学 ， 临床检验诊断学， 2003， 硕士

【摘要】 目的 观察不同浓度牛磺酸对脂质过氧化损伤的红细胞以及自然衰老红细胞的抗氧化酶类和膜骨架蛋白保护作用，以探讨其抗氧化和抗衰老的作用机制。 方法 ①氧化损伤部分：采健康成人静脉血制成红细胞悬液，分为对照组、单纯性氧化损伤组和不同浓度（分别为50mmol／L、150mmol／L、250mmol／L）的牛磺酸保护组。37℃孵育30min后，一起加入过氧化氢（H₂O₂，100mmol／L），孵育后测定红细胞溶血度、红细胞膜丙二醛（MDA）浓度、超氧化物歧化酶（SOD）活性、Na⁺，K⁺-ATPase和Ca²⁺，Mg²⁺-ATPase活性，同时利用扫描电镜和透射电镜观察红细胞形态和红细胞骨架蛋白结构变化，并计算异常红细胞百分率。②自然衰老部分：采健康成人静脉血，分为单纯衰老组和不同浓度（分别为50mmol／L、150mmol／L、250mmol／L）牛磺酸保护组，37℃孵育12h、24h、36h后测定红细胞膜MDA浓度、SOD活性、Na⁺，K⁺-ATPase和Ca²⁺，Mg²⁺-ATPase活性。 结果 ①氧化损伤部分：牛磺酸能明显降低红细胞溶血度，且随着浓度增加其作用逐渐加强，与对照组比较差异有显著意义（F=91.13，q=6.74～22.08，P＜0.01）。在H₂O₂作用下，红细胞膜MDA明显增加、SOD活性、Ca²⁺，Mg²⁺-ATPase活性明显降低，与对照组比较差异有极显著性（F=50.49～93.46，P＜0.001）。牛磺酸各浓度之间的差别亦有显著性（q=2.84～24.31，P＜0.05，0.01）。红细胞膜Na⁺，K⁺-ATPase活性明显降低，与对照组比较差异有极显著性（F=62.57，q=15.36～19.24，P＜0.01），但牛磺酸各浓度之间差别无统计学意义（q=0.74～3.88，P＞0.05）。扫描电镜下，对照组偶见棘红细胞，氧化损伤组棘红细胞百分率为（80±5）％，牛磺酸保护组棘红细胞百分率分别为（72±4）％，（60±6）％，（52±7）％，与氧化损伤组比较差异有显著性（t=5.88～7.90，P＜0.01），且随着牛磺酸浓度增高，棘红细胞百分率逐渐减低。透射电镜下，氧化损伤后红细胞骨架结构破坏严重。预先加入牛磺酸的红细胞骨架结构 中文摘要部分完整。②自然衰老部分:随孵育时间的延长，红细胞膜MDA逐渐增加、SOD活性、Na+，K+一ATPase活性、CaZ+，MgZ+一ATPase活性逐渐降低，与对照组比较差异有显著意义（F一24.83²43.01，尸<0.001）。随牛磺酸浓度的升高，红细胞膜MDA降低、SOD活性、Na+，K+一ATPase活性、c扩+，MgZ十一AT-Pase活性升高逐渐明显，但与对照组比较差异仍有显著意义（F一2.97一57.98，P<0 .001）。 结论牛磺酸在红细胞膜水平能直接对杭氧自由墓引发的脂质过氧化损伤，保护红细胞抗氧化酶类和红细胞膜骨架蛋白结构。另外，牛磺酸能提高红细胞的抗氧化能力，保护红细胞膜骨架结构，减缓红细胞的老化过程。更多还原

【Abstract】 Objective To observe the effect of taurine (Tau) on erythrocyte lipid peroxidative in-jury and erythrocyte survival, and to explore the protective mechanism of Tau on erythro-cyte.Methods (1)Lipid peroxidative injury: Intravenous blood samples taken from healthy adultes were prepared for red blood cell(RBC) suspensions and individed equally into 5 groups:group I for control, groupHwith hydrogen peroxide (H2O2,100mmol/L) induced injury, and group IIIIV V with the same injury as the group II but being pretreated with 3 different concentrations of Tau (50mmol/L, 150mmol/L,250mmol/L), respectively. (2) Senescent erythrocyte: Intravenous blood samples taken from healthy adultes were individ-ed equally into 4 groups:group I for control, group II , ffl , IV were respectively pretreated with 3 different concentrations of Tau (50mmol/L, 150mmol/L, 250mmol/L) , incubated under 37℃ for 12h,24h,36h as erythrocyte normal aging. Hemolytic degree, the concen-tration of erythrocyte membrane lipid peroxdation malondialdehyde (MDA) and the activi-ty of superoxide dismutase (SOD) and ATPase in RBC suspensions after incubation were measured. At the same time, morphology and membrane skeleton structure of erythrocyte was studied with scan electron microscope and transmission electron microscope for eryth-rocytes of lipid injury, respectively.Result After lipid peroxidate injurying, the hemolytic degree of groups protected by taurine decrease significantly(F=91.13,q = 6. 74-22. 08, P<0. 01), and MDA concen-tration also decrease significantly while the activity of SOD and Ca2+ , Mg2+ -ATPase in-creased in the groups that were pretreated with Tau as compared with the control group (F=50. 49 ?93. 46,P<0. 001). Significance can be seen within the groups pretreated with 3 different concentrations of Tau(q=2. 84 - 24. 31, P<0. 05,0. 01). The activity of Na+ , K+-ATPase increased in the groups that were pretreated with Tau as compared with the control group(F=62. 57,g=15. 36 ?19. 24, P<0. 01)while significance cannot be seen within 3 different concentrations of Tau(g=0. 74 - 3. 88, P>0. 05). After normal aging, the hemolytic degree and MDA concentration decreased significantly while the activity of SOD and ATPase increased in the groups that were pretreated with Tau as compared with the control group. Significance can be seen within the groups pretreated with 3 differentconcentrations of Tau (F=24. 83 - 243. 01, P<0. 001),and also can been seen among dif-ferent aging time (F=2. 97-57. 98, P<0. 001). Obvious more erythrocytes with mor-phological abnormality like acanthocytes were seen injurying groups and groups pretreated with 3 different concentrations of Tau comparing the control group under scan electron mi-croscope, and difference has been seen about acanthocyte percentage among the three con-centration Tau-treated groups(t=5. 88- 7. 90, P<0. 01). The erythrocyte membrane skeleton structure of control groups was loose, blurred and partially deficient after injur-ying. Groups pretreated Tau became better.Conclusion Tau could directly counteract the erythrocyte membrane lipid peroxida-tive injury caused by free radicals and aging.更多还原