摘要
目的构建过氧化氢(H2O2)诱导小鼠睾丸间质细胞(TM3)氧化应激模型,为肥胖相关雄性生殖功能低下机制研究提供参考。方法分别以不同浓度H2O2处理TM3细胞4 h,采用噻唑蓝法测定细胞活力,酶联免疫吸附试验检测睾酮水平,比色法测定细胞内活性氧(ROS)含量、总抗氧化能力(TAOC)、超氧化物歧化酶(SOD)活性和过氧化氢酶(CAT)活性。结果 300、600μmol/L H2O2组细胞存活率[(86.7±4.3)%、(46.3±2.0)%]均低于对照组[(100.0±1.7)%](P<0.05);与对照组[(250.09±26.70)pg/m L]比较,150、300、600μmol/L H2O2组TM3细胞睾酮水平[(188.93±7.06)、(179.23±6.66)、(131.59±11.26)pg/m L]均明显降低(P<0.05);与对照组比较,150、300、600μmol/L H2O2组TM3细胞SOD和CAT活性[分别为(2.42±0.17)、(2.69±0.30)、(0.44±0.41)和(56.76±12.17)、(23.45±2.17)、(24.55±18.05)U/mg protein]均明显下降(P<0.05)。结论成功构建以H2O2为诱导剂的TM3细胞体外氧化应激模型。
Objective To establish an oxidative damage model with hydrogen peroxide（ H2O2） in mouse Leydig（ TM3） cells for studying possible mechanism of decreased reproductive function in obese males. Methods After 4hours＇ treatment with different concentration of H2O2,the cell viability was determined with 3-（ 4,5-dimethylthiazol-2-yl）-2,5-diphenyl tetrazolium bromide（ MTT） method; the testosterone level was determined with enzyme-linked immunosorbent assay（ ELISA）; the contents of reactive oxygen species（ ROS）,total antioxidant capacity（ TAOC） and activities of superoxide dismutase（ SOD） and catalase（ CAT） were detected with spectrophotometry. Results The viability of TM3 cells（ 86. 7 ± 4. 3% and 46. 3 ± 2. 0%） of 300 and 600 μmol / L H2O2 exposure groups decreased significantly compared to that of the control group（ 100 ± 1. 7%）（ all P〈0. 05）. The testosterone level（ 188. 93 ± 7. 06,179. 23 ± 6. 66,and 131. 59 ± 11. 26 pg / m L） of 150,300,600 μmol / L H2O2 exposure groups were significantly lower than that of the control group（ 250. 09 ± 26. 70 pg / m L）（ all P〈0. 05）. The activity of SOD（ 2. 42 ± 0. 17,2. 69 ± 0. 30,and 0. 44 ± 0. 41 U / mg protein） and CAT（ 56. 76 ± 12. 17,23. 45 ± 2. 17,and 24. 55 ± 18. 05 U / mg protein） of 150,300,and 600 μmol / L H2O2 exposure groups decreased significantly compared to those of the control group（ all P〈0. 05）. Conclusion The TM3 cell model of oxidative injury was established successfully with the administration of H2O2.
出处
《中国公共卫生》
CAS
CSCD
北大核心
2015年第3期312-314,共3页
Chinese Journal of Public Health
基金
国家自然科学基金青年科学基金(81202227)
辽宁省教育厅一般项目(L2012365)
