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目的探讨氧化应激在习惯性流产致不孕发病机制中的作用。方法选择2013年1月—2014年6月收治的46例习惯性流产致不孕患者作为观察组,同期选择健康已育非妊娠女性40例作为对照组。清晨抽取受试者空腹静脉血,血样使用枸橼酸钠抗凝,2 500 r/min离心10 min,分离红细胞,制备红细胞膜,溶血样本弃用。黄嘌呤氧化酶法检测两组受试者红细胞膜匀浆上清液中氧化应激标志物超氧化物歧化酶(superoxide dismutase,SOD)活性;还原法检测红细胞膜谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)活性;硫代巴比妥酸法测定红细胞膜匀浆上清液中脂质过氧化物丙二醛(malondialdehyde,MDA)含量;以上检测均以考马斯亮蓝法检测红细胞膜蛋白含量,严格按照试剂盒说明书操作。酶促反应法测定红细胞膜Na+-K+-ATP酶活力,H2O2诱导氧化法检测红细胞溶血度。计量资料采用t检验,P<0.05为差异有统计学意义。结果观察组红细胞膜SOD、GSH-Px、Na+-K+-ATP酶水平[(11.62±3.21)、(33.56±5.15)U/ml、(0.45±0.07)U/(mg·prot)]较对照组[(23.70±4.09)、(57.84±6.18)U/ml、(0.63±0.08)U/mg]低,差异均有统计学意义(均P<0.05)。观察组MDA、红细胞溶血度[(2.28±0.44)nmol/ml、(47.63±7.99)%]较对照组[(0.68±0.14)nmol/ml、(40.60±4.89)%]高,差异均有统计学意义(均P<0.05)。结论体内氧化应激增强,抗氧化能力减弱,机体及生殖系统受到氧化应激损伤可能是习惯性流产造成不孕的原因之一。
Objective To investigate the role of oxidative stress in the pathogenesis of infertility caused by habitual abortion. Methods Forty-six patients with habitual abortion-induced infertility who were admitted to our hospital from January 2013 to June 2014 were selected as observation group and 40 healthy non-pregnant women were selected as control group in the same period. Fasting venous blood samples were taken early in the morning, the blood samples were anticoagulated with sodium citrate, centrifuged at 2500 r / min for 10 min, the erythrocytes were separated, and the erythrocyte membrane was prepared. The hemolysis samples were discarded. The activity of superoxide dismutase (SOD), an oxidative stress marker in erythrocyte membrane homogenate supernatant, was detected by xanthine oxidase method. The activity of glutathione peroxidase glutathione peroxidase (GSH-Px) activity. The content of malondialdehyde (MDA) in supernatant of erythrocyte membrane homogenate was measured by thiobarbituric acid method. Erythrocyte membrane protein content, in strict accordance with kit instructions. Enzyme reaction assay of erythrocyte membrane Na + -K + -ATP enzyme activity, H2O2 induced oxidation of hemolysis of erythrocytes. Measurement data using t test, P <0.05 for the difference was statistically significant. Results Compared with the control group, the levels of SOD, GSH-Px and Na + -K + -ATPase in erythrocyte membrane in the observation group were significantly higher than those in the control group [(11.62 ± 3.21), (33.56 ± 5.15) U / ml, [(23.70 ± 4.09), (57.84 ± 6.18) U / ml, (0.63 ± 0.08) U / mg] respectively. There were significant differences between the two groups (all P <0.05). The MDA in the observation group was significantly higher than that in the control group [(2.28 ± 0.44) nmol / ml, (47.63 ± 7.99)%, [(0.68 ± 0.14) nmol / ml, (40.60 ± 4.89)%] Significance (both P <0.05). Conclusions Increased oxidative stress, decreased antioxidant capacity, and oxidative stress in the body and the reproductive system may be one of the causes of infertility caused by habitual abortion.