银杏黄酮对环磷酰胺所致睾丸毒性的保护作用的实验研究

The protective effect of ginkgetin in impairment to testicular toxicity by cyclophosphamide

目的:在先期研究已证实环磷酰胺(Cyclophosphamide,CP)代谢产物丙烯醛(Acrolein,ACR)的氧化应激作用损伤支持细胞是CP所致睾丸损伤的原因之一的基础上探讨具有抗氧化作用的银杏黄酮对其损伤的保护作用。方法:以原代培养的新生Wistar大鼠支持细胞为模型,实验组分为1)添加银杏黄酮孵育细胞12h后加入ACR组;2)银杏黄酮、ACR同时加入组;3)银杏黄酮和ACR孵育12h后再加入细胞组。对照组分为添加维生素E孵育细胞12h后分别加入ACR组,仅添加ACR的阴性对照组及不添加任何处理因素的阳性对照组。各组3h后用MTT法测定细胞活性,考马斯亮蓝法测定蛋白浓度;硫代巴比妥法和Fenton法检测细胞丙二醛(MDA)、羟自由基(OH.)含量,三价铁还原法、黄嘌呤氧化酶法和NADPH还原法测定细胞总抗氧化能力(T-AOC)、超氧化物歧化酶(SOD)活性和谷胱甘肽还原酶(GR)活性。结果:银杏黄酮与ACR同时加入细胞组和银杏黄酮与ACR孵育12h后同时加入组较阴性对照组各测量参数无显著性差异(P>0.05)。而提前添加银杏黄酮组较阴性对照组各参数均有显著性差异:1)细胞活力较阴性对照组显著升高(P<0.05);2)MDA和T-ACO含量较阴性对照组明显降低(P<0.05);3)T-AOC、SOD和GR活性较阴性对照组明显升高(P<0.05)。且银杏黄酮上述抗氧化作用明显强于Vit E组(P<0.05)结论:CP代谢产物ACR在对未成熟睾丸支持细胞产生氧化应激损害时,银杏黄酮可以提高支持细胞的抗氧化酶活性,清除氧自由基,对支持细胞具有明显的保护作用。为进一步探讨CP致睾丸损伤的防护措施奠定了基础。

Objective：To investigate the protective effect of ginkgetin from oxidative stress on immature testis induced by cyclophosphamide （CP） metabolite acrolein （ACR） in vitro. Methods：The sertoli cells were isolated from 7-days old SD rats＇ testes,culture and passage to F2 generations. Experiment groups were divided into： 1）Ginkgetin incubated for 12 hours and then treat with ACR 2） Ginkgetin and ACR add to cells at the same time 3） Ginkgetin and ACR were incubated for 12 hours,then add to cells. Control group include： 1） Vitamin E were incubated cells for 12 hours, then add ACR respectively 2） Added ACR only as the negative control group 3） Not add any factors as the positive control group. 3 h later the viability of sertoli cell were determined by MTT, Protein concentration was determinate by Coomassie Brilliant Blue. Malondialdehyde （MDA）, hydroxyl radical （OH）, total antioxidant capacity（T-ACO）, Superoxide dismutase（SOD） and glutathione reductase （GR） activity were tested, Results： 1 ） Ginkgetin（add before ACR） can significantly raise the viability of the cells（P〈0.05）. 2） Ginkgetin can degrade MDA and OH of cells that treated by acrolein significantly（P〈0.05）. 3） Ginkgetin raised Total antioxidant,GR and SOD activity of sertoli cells treated by acrolein significantly（P〈0.05）. Conclusion：Ginkgetin can protect sertoli cells of immature testes from ACR oxidative injury in vitro.