大豆异黄酮对环磷酰胺损伤雄性小鼠生殖功能的修复作用

Effect of Soy Isoflavones on Repairing Reproductive Function Damage of Male Mice Induced by Cyclophosphamide

为探明大豆异黄酮(SI)对环磷酰胺(CTX)所致雄性小鼠生殖功能损伤的修复作用及其机制,本试验将小鼠随机分为空白对照组、模型组(CTX组)、SI低、中、高剂量组[100 mg/(kg·bw)、200 mg/(kg·bw)、400 mg/(kg·bw)]和甲睾酮(MT)对照组,除空白对照组外,其余各组腹腔注射CTX[50 mg/(kg·bw)]连续7 d,构建雄性小鼠体内生精障碍模型,造模第2天开始,SI各剂量组和MT对照组分别灌胃给药,连续30 d后,测定雄性小鼠的睾丸指数、精子密度、精子畸变率;检测血清和睾丸组织中丙二醛(MDA)含量;检测睾丸组织匀浆中琥珀酸脱氢酶(SDH)、碱性磷酸酶(AΚP)、Na^(+)-Κ^(+)-ATP酶、NF-κB、TNF-α水平;检测睾丸组织总蛋白样本中NF-κB、COX2和iNOS蛋白的表达水平。结果显示,SI各剂量组与MT对照组效果相当;与CTX组相比,SI各剂量组小鼠精子密度显著升高而精子畸变率显著降低,睾丸组织中SDH、AKP和Na^(+)-K^(+)-ATP酶活性均显著升高(P<0.05或P<0.01);SI中、高剂量组小鼠血清和睾丸组织中MDA含量显著降低(P<0.05),睾丸组织中TNF-α和NF-κB的含量显著下降(P<0.05或P<0.01);与空白对照组比较,CTX组睾丸组织中NF-κB、COX2和iNOS蛋白的表达明显增加,与CTX组相比,SI高剂量组对蛋白表达有明显抑制作用(P<0.05或P<0.01)。结果表明,SI对CTX引起的小鼠生精功能障碍有一定修复作用,其作用机制可能与抑制NF-κB/COX2/iNOS氧化应激通路有关。

This study examined the effect of soy isoflavone(SI)on repairing reproductive function damage induced by cyclophosphamide(CTX)in male mice and preliminarily discussed its mechanism.Mice were randomly divided into blank control group(BC group),model group(CTX group),SI low-dose,medium-dose and high-dose group[100 mg/(kg·bw),200 mg/(kg·bw)and 400 mg/(kg·bw)],and methyltestosterone control group(MT group).CTX[50 mg/(kg·bw)]was injected intraperitoneally for 7 days to establish a spermatogenesis disorder model in male mice.From the second day,different concentrations of SI were given by intragastric administration.After 30 days,the testicular index,sperm density,and sperm aberration rate of male mice were measured;the contents of malondialdehyde(MDA)in serum and testicular tissue of mice were detected by kits;further,the activities of SDH,AΚP and Na+-Κ+-ATPase in the testicular tissue of mice were detected by kits;specifically,the TNF-αand NF-κB in the testicular tissue of mice were detected by ELISA;the expression level of NF-κB,COX2 and iNOS protein were detected by Western blot.The results showed that the SI dose groups had comparable effects to the MT group.Compared with the CTX group,the sperm density of mice,and the activities of SDH,AΚP and Na+-Κ+-ATPase in the testicular tissue of mice were increased,and sperm aberration rate was decreased in the SI dose groups(P<0.05 or P<0.01),the MDA content in serum and testicular tissue of mice,and TNF-αand NF-κB in mouse testis were decreased in the SI medium-dose group and SI high-dose group(P<0.05 or P<0.01).Additionally,compared with the BC group,the expression of NF-κB,COX2,and iNOS proteins in the testicular tissue was visibly increased in the CTX group(P<0.05 or P<0.01).In contrast,compared with the CTX group,the SI high-dose group inhibited protein expression with statistical differences(P<0.05 or P<0.01).Thus,SI have preventive effect on the reproductive system dysfunction of male mice induced by CTX,and its mechanism of action is related to the inhibition of NF-κB/COX2/iNOS oxidative stress pathway.