铁过载对性成熟期雄性大鼠生育能力的影响

doi:10.3969/j.issn.1004-616x.2017.02.012

摘要/Abstract

摘要：

目的：研究铁过载对性成熟期雄性大鼠生育能力的影响。方法：6~7周龄SPF级雄性Wistar大鼠40只，按体质量随机分为对照组、缺铁组、低剂量铁过载组及高剂量铁过载组，每组10只，经腹腔注射含Fe2+分别为0.9、0.3、9和18 mg的右旋糖酐铁注射液，隔日注射1次，连续6周。于第6周末经腹主动脉采血，采用比色法检测血清铁含量。留取睾丸及附睾，计算脏器指数，制备精子悬液，显微镜下观察精子数量、精子运动情况及精子畸形情况。采用可见光法检测睾丸组织中过氧化氢酶（CAT）水平，黄嘌呤氧化酶法检测睾丸组织中总超氧化物歧化酶（T-SOD）水平，比色法检测睾丸组织中铁含量及谷胱甘肽过氧化物酶（GSH-Px）水平，硫代巴比妥酸法检测睾丸组织中丙二醛（MDA）水平。结果：缺铁组大鼠血清铁含量明显低于对照组（P < 0.01），而睾丸组织中铁含量与对照组比较，差异无统计学意义（P > 0.05）。低剂量和高剂量铁过载组大鼠血清和睾丸组织中的铁含量均显著高于对照组（P < 0.01）。与对照组相比，缺铁组大鼠的精子数量、精子活动率及精子畸形率的差异均无统计学意义（P > 0.05）。低剂量铁过载组大鼠的精子活动率显著低于对照组（P < 0.01）。高剂量铁过载组大鼠的精子数量和精子活动率显著低于对照组和低剂量铁过载组，且精子畸形率显著高于对照组和低剂量铁过载组（P < 0.05）。缺铁组大鼠睾丸GSH-Px活性显著低于对照组（P < 0.05）。低剂量铁过载组大鼠睾丸GSH-Px活性显著低于对照组，而MDA水平显著高于对照组（P < 0.05）。高剂量铁过载组大鼠睾丸CAT水平及GSH-Px活性均显著低于对照组和低剂量铁过载组，而MDA水平显著高于对照组和低剂量铁过载组（P < 0.05）。结论：铁过载可影响性成熟期雄性大鼠的生育能力，表现为精子数量减少、精子运动率降低以及精子畸形率升高，其原因可能与铁过载所导致的高氧化应激水平有关。

关键词: 铁过载, 精子数量, 精子活动率, 精子畸形率, 氧化应激

Abstract:

OBJECTIVE: The aim of the study was to observe the effect of iron overload on fertility of sexually mature male rats. METHODS: According to bodyweights,40 healthy male Wistar rats were randomly divided into four groups:control,iron deficiency,low-dose iron overload and high-dose iron overload groups. All rats were injected intraperitoneally every other day. The content of the Fe2+ in these four groups were 0.9,0.3,9 and 18 mg,respectively. After 6 weeks,blood samples from these rats were collected to determine serum iron levels. Testes and epididymides were removed and weighed. Sperm numbers were counted. Sperm mobility and teratosperm rates were calculated. Iron concentration,catalase (CAT),total superoxide dismutase (T-SOD),glutathione peroxidase (GSH-Px) and malonaldehyde (MDA) levels in the testes were determined. RESULTS: Compared with the control group,the serum iron concentration of the iron deficiency group was significantly lower (P < 0.01). However,there was no significant difference in the iron concentrations of the testes (P > 0.05). The serum and testes iron concentrations in the low- and high-doses iron overload groups were both significantly higher than that in the control group (all P < 0.01). There was no significant difference in the sperm count,the sperm motility rate and the teratosperm rate between the iron deficiency group and the control group (all P > 0.05). Compared with the control group,the sperm motility rate in the low-dose iron overload group was significantly decreased (P < 0.01);the sperm count and the sperm motility rate in the high-dose iron overload group were all significantly lower than those of control group and low-dose iron overload group,but the teratosperm rate was significantly higher than that of control group and low-dose iron overload group (all P < 0.05). The rats in the iron deficiency group had significantly lower GSH-Px levels in the testis comparing with the control group. Moreover,compared with the control group,the GSH-Px levels in the testis of the low-dose iron overload group were significantly lower,but the MDA level was significantly higher (all P < 0.05). The CAT and GSH-Px levels in the testis of the high-dose iron overload group were all significantly lower than those of control group and low-dose iron overload group,but the MDA level was significantly higher than that of control group and low-dose iron overload group (all P < 0.05). CONCLUSION: Iron overload can affect the fertility of sexually mature male rats,including reduced sperm count,decreased sperm motility rate and increased teratosperm rate. These adverse effects may be related to the increased oxidative stress level caused by iron overload.

Key words: iron overload, sperm count, sperm motility rate, teratosperm rate, oxidative stress

中图分类号:

R321.1

张华琦, 马爱国, 孙永叶, 钟凤, 蔡静, 何丽敏. 铁过载对性成熟期雄性大鼠生育能力的影响[J]. 癌变·畸变·突变, 2017, 29(2): 134-138,142.

ZHANG Huaqi, MA Aiguo, SUN Yongye, ZHONG Feng, CAI Jing, HE Limin. Effect of iron overload on fertility of sexually mature male rats[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2017, 29(2): 134-138,142.

参考文献

[1] ZHANG A S,ENNS C A. Iron homeostasis:Recently identified proteins provide insight into novel control mechanisms [J]. J Biol Chem,2009,284(2):711-715．

[2] KARTIKASARI A E,GCORGION N A,VISSEREN F L,et al. Endothelial activation and induction of monocyte adhesion by nontransferrin-bound iron present in human sera[J]. FASEB J,2006,20(2):353-355.

[3] 赵长峰,杨洪旗,姜会敏,等．铁缺乏对孕鼠及其胎鼠体内元素分布的影响[J]．卫生研究,2001,30(5):277-279．

[4] MEANS R J. Advances in the anemia of chronic disease[J]. Int J Hematol,1999,70:7-12.

[5] ANDREWS N C. Medical progress:disorders of iron metabolism[J]. N Engl J Med,1999,341:1986-1995．

[6] NICOLAS G,VIAAE L,LOU D Q,et al. Constitutive hepcidin expression prevents iron overload in a mouse model of hemochromatosis[J]. Nat Genet,2003,34:97-101.

[7] 张梦雪,马爱国,胡迎芬,等．铁摄入过量对大鼠胚胎发育营养的实验研究[J] ．中国食物与营养,2014,20(2):65-69．

[8] 王心如. 毒理学基础[M]. 北京:人民卫生出版社,2003:119．

[9] HAMETNER S,WIMMER I,HAIDER L,et al. Iron and oxidative damage in the multiple sclerosis brain[J]. J Neurol Sci,2013,333(11):386-387.

[10] 李新华,张欣欣. 铁负荷过多综合征的研究进展—2007 年美国肝病学会 (AASLD) 年会专题报道[J]. 肝脏,2008,13(2):148-150.

[11] MIRA MALKOV,YONIT FISHER,JEREMY DON. Developmental Schedule of the postnatal rat testis determined by flow cytometry[J]. Biol Reprod,1998,59:84-92.

[12] 陈绍坤,余红,李洁,等. 微量元素与男性不育症患者精子质量的关系分析[J]. 现代医药卫生,2012,28(6):849-850.

[13] 何丽敏,孙永叶,蔡静,等. 铁过量对大鼠淋巴细胞DNA损伤的影响[J]. 癌变·畸变·突变,2015,27(1):59-63.

[14] 王亚锦,胡迎芬,王洋,等. 铁过载对大鼠骨髓及肝脾组织的影响[J]. 现代生物医学进展,2015,15(19):3641-3644.

[15] 徐谊朝,朱伟杰,李菁,等. 有生育需求中老年男性精子参数的评价[J]. 生殖与避孕,2014,34(007):599-602.

[16] NASRI S,REZAI-ZARCHI S,KERISHCHI P,et al. The Effect of iron oxide nanoparticles on sperm numbers and mobility in male mice[J]. Zahedan J Res Med Sci,2015:29-31.

[17] JALILI C,SALAHSHOOR M R,NASERI A. Protective effect of Urtica dioica L against nicotine-induced damage on sperm parameters,testosterone and testis tissue in mice[J]. Iran J Reprod Med,2014,12(6):401-408.

[18] WANG J,PANTOPOULOS K. Regulation of cellular iron metabolism[J]. Biochem J,2011,434(1):365-381.

[19] 陈玉明,陈杰,王日文,等. Fe²⁺对雄性泥鳅生殖毒性的研究[J]. 湖北农业科学,2011,50(7):1444-1447.

[20] FINAZZI D,AROSIO P. Biology of ferritin in mammals:an update on iron storage,oxidative damage and neurodegeneration [J]. Arch Toxicol,2014,88(10):1787-1802.

[21] 李云春,马爱国,汪求真,等. 叶黄素对铁过载大鼠氧化应激水平影响的实验研究[J]. 中国食物与营养,2014,20(3):58-61.

[22] MOJICA-VILLEGAS M A,IZQUIERDO-VEGA J A,CHAMORRO-CEVALLOS G,et al. Protective effect of resveratrol on biomarkers of oxidative stress induced by iron/ascorbate in mouse spermatozoa[J]. Nutrients,2014,6(2):489-503.

[23] WELLEJUS A,POULSEN H E,LOFT S. Iron-induced oxidative DNA damage in rat sperm cells in vivo and in vitro[J]. Free Radic Res,2000,32(1):75-83.

[24] MARZEC-WRÓBLEWSKA U,KAMI?SKI P,?AKOTA P,et al. Zinc and iron concentration and SOD activity in human semen and seminal plasma[J]. Biol Trace Elem Res,2011,143(1):167-177.

[25] MATÉS J M,PÉREZ-GÓMEZ C,NUNEZ D C. Antioxidant enzymes and human diseases[J]. Clin Biochemi,1999,32(8):595-603.