叶酸对炔雌醚致雄性大鼠生殖损伤的保护作用研究

doi:10.11843/j.issn.0366-6964.2017.02.019

畜牧兽医学报 ›› 2017, Vol. 48 ›› Issue (2): 351-358.doi: 10.11843/j.issn.0366-6964.2017.02.019

叶酸对炔雌醚致雄性大鼠生殖损伤的保护作用研究

李健¹，陈福宁²，陈耀星³，王子旭³，董玉兰³，曹静³，司丽芳¹，刘志军¹，赵战勤^1*

（1.河南科技大学动物科技学院，洛阳471023；2.北京昌平区中西医结合医院，北京102208；3.中国农业大学动物医学院，北京100193）

收稿日期:2016-08-11 出版日期:2017-02-23 发布日期:2017-02-23
通讯作者: 赵战勤，博士，副教授，Tel:0379-64282341,E-mail：zhaozhanqin@126.com
作者简介:李健（1980-），男，安徽亳州人，博士，副教授，主要从事生殖毒理学、动物解剖学与组织胚胎学教学与研究，E-mail：lijian800702@126.com
基金资助:
国家自然科学基金项目（U1404325；31302106）；河南省教育厅高等学校重点科研项目基础研究计划（16A230004）；河南科技大学青年科学基金项目（2015QN032）；河南科技大学高级别科研项目培育基金（序号4）；河南省自然科学基金项目（162300410081）

The Study of Protection of Folacin against Quinestrol Induced Reproductive Toxicity in Male Rat

LI Jian¹, CHEN Fu-ning², CHEN Yao-xing³, WANG Zi-xu³, DONG Yu-lan³, CAO Jing³,SI Li-fang¹,LIU Zhi-jun¹,ZHAO Zhan-qin^1*

(1.College of Animal Science and Technology, Henan University of Science and Technology, Luoyang 471023, China; 2.Changping Hospital of Integrated Chinese and Western Medicine, Beijing 102208, China 3. College of Veterinary Medicine, China Agricultural University, Beijing 100193, China)

Received:2016-08-11 Online:2017-02-23 Published:2017-02-23

摘要/Abstract

摘要：

旨在探讨叶酸对炔雌醚致雄性大鼠生殖损伤的保护作用。将20只8周龄成年雄性SD大鼠适应饲养1周后，随机分为4组。对照组：橄榄油+生理盐水；炔雌醚组：1.0 mg•kg^-1炔雌醚；炔雌醚+叶酸组：1.0 mg•kg^-1炔雌醚+1.1 mg•kg^-1叶酸；叶酸组：1.1 mg•kg^-1叶酸；灌胃处理，每天一次，连续2周。处理结束后，取生殖器官称重，检测精子质量，测定睾丸抗氧化功能，制作睾丸组织切片，观察睾丸组织结构，通过免疫组织化学染色检测Caspase-3与eNOS表达。结果显示，炔雌醚单独处理组造成大鼠生殖器官萎缩，重量下降，精子发生异常，精子质量下降，抗氧化酶活性下降，睾丸发生氧化应激；与之相反，凋亡效应酶Caspase-3与eNOS表达增加。炔雌醚+叶酸处理组通过减少eNOS表达降低氧化应激，抑制Caspase-3表达，降低精子畸形率、生精小管损伤，最终改善雄性大鼠生殖功能。与对照组相比，单独叶酸处理组睾丸抗氧化酶活性、组织结构和精子质量均有一定程度提高，无显著差异。本研究首次证实叶酸有效地抑制了炔雌醚对雄性大鼠生殖功能的破坏作用。

Abstract:

The study was conducted to investigate the effects of folacin on quinestrol induced abberant reproduction in adult male rat. Twenty 8-week-old adult male SD rats were randomly divided into 4 groups after 1-week of adaptive feeding. Control group: olive oil + physiological saline; Quinestrol group: 1.0 mg•kg^-1 quinestrol; Quinestrol + folacin group: 1.0 mg•kg^-1 quinestrol + 1.1 mg•kg^-1 folacin (dissolved in saline); Folacin group: 1.1 mg•kg^-1 folacin. Quinestrol and folacin were given intragastric administration to male rats daily for 2 weeks. After the treatments, the sexual organs were weighed, semen quality were tested, antioxidant status of testes were investigated, testis tissue section were made, histopathology of testes were observed, the expressions of Caspase-3 and endothelial nitric oxide synthase (eNOS) were detected by immunohistochemical staining. The results showed that atrophy of sexual organs were induced, the weights of sexual organs were decreased, abnormal spermatogenesis were abberant, sperm quality were reduced, antioxidant enzyme activity were decreased, oxidative stress were happend in testes after the treatment of quinestrol alone. By contrast, the expressions of Caspase-3 and eNOS in the testes were increased. In the group of quinestrol and folacin treatment, oxidative stress were reduced by decreasing eNOS expression, the expression of Caspase-3 were inhibited, the sperm deformity rate and the damage of the tubule were reduced, the reproductive function of male rats were improved significantly. Compared with the control group, the testicular antioxidant enzyme activities, tissue structure and sperm quality were ameliorated to some extent in the folacin treatment group, there was no significant difference. The results clearly demonstrate for the first time that folacin effectively inhibited the damaging effects of quinestrol on reproductive function in adult male rats.

中图分类号:

S85
S814

李健，陈福宁，陈耀星，王子旭，董玉兰，曹静，司丽芳，刘志军，赵战勤. 叶酸对炔雌醚致雄性大鼠生殖损伤的保护作用研究[J]. 畜牧兽医学报, 2017, 48(2): 351-358.

LI Jian, CHEN Fu-ning, CHEN Yao-xing, WANG Zi-xu, DONG Yu-lan, CAO Jing,SI Li-fang,LIU Zhi-jun,ZHAO Zhan-qin. The Study of Protection of Folacin against Quinestrol Induced Reproductive Toxicity in Male Rat[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(2): 351-358.

参考文献

［1］TANG T, QIAN K, SHI T, et al. Photodegradation of quinestrol in waters and the transformation products by UV irradiation［J］. Chemosphere, 2012. 89:1419-1425.
［2］ZHAO M, LIU M, LI D, et al. Anti-fertility effect of levonorgestrel and quinestrol in Brandt’s voles (Lasiopodomys brandtii) ［J］. Integr Zool, 2007, 2(4):260-268.
［3］LÜ X, GUO Y, SHI D. Effects of quinestrol on reproductive hormone expression, secretion, and receptor levels in female Mongolian gerbils (Meriones unguiculatus) ［J］.Theriogenology, 2012, 77(6):1223-1231.
［4］SHEN W, SHI D, WANG D, et al. Inhibitive effects of quinestrol on male testes in Mongolian gerbils (Meriones unguiculatus) ［J］. Res Vet Sci, 2012, 93(2): 907-913.
［5］LI J, CHEN F, LI C, et al. Quinestrol induces spermatogenic apoptosis in vivo via increasing pro-apoptotic proteins in adult male mice［J］. Tissue Cell, 46(5): 318-325.
［6］HEMACHAND T, SHAHA C. Functional role of sperm surface glutathione S-transferases and extra cellular glutathione in the haploid spermatozoa under oxidative stress［J］.FEBS Lett, 2003, 538(1):14-18.
［7］AGARWAL A, SALEH R A, BEDAIWY M A. Role of reactive oxygen species in the pathophysiology of human reproduction［J］. Fertil Steril, 2003, 79(4):829-843.
［8］SUBUDHI U, DASA K, PAITAL B, et al. Alleviation of enhanced oxidative stress and oxygen consumption of L-thyroxine induced hyperthyroid rat liver mitochondria by vitamin E and curcumin［J］. Chem Biol Interact, 2008, 173(2):105-114.
［9］BURTON G J, JAUNIAUX E. Oxidative stress［J］. Best Pract Res Clin Obstet Gynaecol, 2011, 25(3):287-299.
［10］VAANHOLT L M, MILNE A, ZHENG Y, et al. Oxidative costs of reproduction: Oxidative stress in mice fed standard and low antioxidant diets［J］. Physiol Behav, 2016, 154:1-7.
［11］PLANTAMP C, SALORT K, GIBERT P, et al. All or nothing: Survival, reproduction and oxidative balance in Spotted Wing Drosophila (Drosophila suzukii) in response to cold［J］. J Insect Physiol, 2016, 8:28-36.
［12］刘艳利，申静，支丽慧，等．叶酸调控鸡脾和胸腺IGF-2表达的表观遗传机制探究［J］．畜牧兽医学报，2016，47(2):296-304.
LIU Y L, SHEN J, ZHI L H, et al. The study on epigenetic mechanism of IGF2 expression in spleen and thymus regulated by folic acid in broilers［J］. Acta Veterinaria et Zootechnica Sinica, 2016, 47(2):296-304.(in Chinese)
［13］KAMEN B. A review on folate and antifolate pharmacology［J］. Semin Oncol, 1997, 24 (5):18-39.
［14］SHALABY M A, EL ZORBA H Y, ZIADA R M. Reproductive toxicity of methomyl insecticide in male rats and protective effect of folic acid［J］. Food Chem Toxicol, 2010, 48(11):3221-3226.
［15］CAILLE N, RODINA M, KOCOUR M, et al. Quantity, motility and fertility of tench Tinca tinca (L.) sperm in relation to LHRH analogue and carp pituitary treatments［J］. Aqua Int, 2006, 14(1):75-87.
［16］SÖNMEZ M, TÜRK G, YÜCE A. The effect of ascorbic acid supplementation on sperm quality, lipid peroxidation and testosterone levels of male Wistar rats［J］.Theriogenology, 2005, 63(7): 2063-2072.
［17］WATSON P F. Use of a Giemsa stain to detect changes in acrosomes of frozen ram spermatozoa［J］. Vet Rec, 1975, 97(1):12-15.
［18］MA A, YANG X, WANG Z, et al. Adult exposure to diethylstilbestrol induces spermatogenic cell apoptosis in vivo through increased oxidative stress in male hamster［J］. Reprod Toxicol, 2008,25:367-373.
［19］JURAND A, GOEL S C. The use of methyl green-pyronin staining after glutaraldehyde fixation and paraffin or araldite embedding ［J］. Tissue Cell, 1976, 8(2):389-394.
［20］李健，陈耀星，陈福宁，等．MAPK 信号通路在炔雌醚诱导大鼠生精细胞凋亡中的作用研究［J］．畜牧兽医学报，2016，47(2): 381-387.
LI J, CHEN Y X, CHEN F N, et al. The role of MAPK signaling pathway in quinestrol-induced apoptosis of spermatogenic cells in rat［J］. Acta Veterinaria et Zootechnica Sinica, 2016, 47(2): 381-387.(in Chinese)
［21］CAI L, CHEN S, EVANS T, et al. Apoptotic germ-cell death and testicular damage in experimental diabetes: prevention by endothelin antagonism［J］. Urol Res, 2000, 28(5):342-347.
［22］CAMERON D F, MURRAY F T, DRYLIE D D. Interstitial compartment pathology and spermatogenic disruption in testes from impotent diabetic men［J］. Anat Rec, 1985, 213(1):53-62.
［23］BAN Y, KOMATSU T, KEMI M, et al. Testicular spermatid and epididymal sperm head counts as an indicator for reproductive toxicity in rats［J］. Exp Anim, 1995, 44(4):315-322.
［24］FENECH M. Micronutrients and genomic stability: a new paradigm for recommended dietary allowances (RDAs) ［J］. Food Chem Toxicol, 2002, 40(8):1113-1117.
［25］PADMANABHAN S, TRIPATHI D N, VIKRAM A, et al. Methotrexate-induced cytotoxicity and genotoxicity in germ cells of mice: Intervention of folic and folinic acid［J］. Mutat Res, 2009, 673(1):43-52.
［26］IBRAHIM W, TOUSSON E, MM ALI E, et al. Folic acid alleviates oxidative stress and hyperhomocysteinemia involved in testicular dysfunction of hypothyroid rats［J］. Gen Comp Endocr, 2011, 174(2) :143-149.
［27］SWAYNE B G, KAWATA A, BEHAN N A, et al. Investigating the effects of dietary folic acid on sperm count, DNA damage and mutation in Balb/c mice［J］. Mutat Res, 2012, 737(1): 1-7.
［28］MYAR C A, INGERSOLL R, WALLOCK L M. Folate levels and the effects of folate deficiency on the reproductive organs of male rats［J］. FASEB J, 1999, 13:229-232.
［29］FORGES T, MONNIER-BARBARINO P, ALBERTO J M, et al. Impact of folate and homocysteine metabolism on human reproductive health［J］. Hum Reprod Update, 2007, 13(3):225-238.
［30］LI J, WANG H, ZHANG J, et al. Abnormal secretion of reproductive hormones and antioxidant status involved in quinestrol induced reproductive toxicity in adult male rat［J］. Tissue Cell, 2014, 46(1): 27-32.
［31］STANGER O. Physiology of folic acid in health and disease［J］. Curr Drug Metab, 2002, 3(2):211-223.
［32］DEGTEREV A, BOYCE M, YUAN J. A decade of caspases［J］. Oncogene, 2003, 22(53):8543-8567.
［33］ZABIHI S, ERIKSSON U J, Wentzel P. Folic acid supplementation affects ROS scavenging enzymes, enhances Vegf-A, and diminishes apoptotic state in yolk sacs of embryos of diabetic rats［J］. Reprod Toxicol, 2007, 23(4):486-498
［34］李运书，孟祥硕，崔策，等．叶酸对胚胎大鼠心肌细胞凋亡及Bax、Bcl-2表达的影响［J］．承德医学院学报，2015, 3(14):259-261.
LI Y S, MENG X S, CUI C, et al. Effect of folic acid on cardiomyocyte apoptosis and the expressions of Bcl-2 and Bax in Embryonic rat［J］. Journal of Chengde Medical College, 2015, 3(14):259-261.(in Chinese)

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叶酸对炔雌醚致雄性大鼠生殖损伤的保护作用研究

The Study of Protection of Folacin against Quinestrol Induced Reproductive Toxicity in Male Rat

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