为研究大豆异黄酮(soybean isoflavones,SI)对香猪睾丸形态及精子发生标志基因表达的影响,选择40头健康的28日龄雄性香猪,随机分成4组,每组10个重复,每个重复1头猪。1、2、3和4组分别在基础饲粮中添加0、125、250和500mg/kg SI,饲喂60d后从各组分别随机选取5头屠宰,采取睾丸样品,分析睾丸的组织形态及E型钙粘连蛋白1(Cdh1)、联会复合体蛋白3(SCP3)、过渡蛋白1(Tnp1)和波形蛋白(Vim)基因的表达。结果表明:各添加水平的SI均造成了睾丸曲细精管中空泡的形成,减少了管腔中成熟精子的数量;饲粮添加SI极显著降低了睾丸组织Cdh1、SCP3和Tnp1基因的表达量(P<0.01);添加250mg/kg的SI显著降低了睾丸组织Vim基因的表达量(P<0.05)。由此可知,饲粮添加125、250、500mg/kg的SI对香猪睾丸形态有明显损伤,抑制了精子发生标志基因的表达;250mg/kg SI显著抑制猪睾丸支持细胞标志基因表达。
To investigate the effects of soybean isoflavones (SI) on testis morphology and expression of marker genes during spermatogenesis of Xiang pigs, 40 healthy male pigs aged 28 days were selected and divided randomly into 4 groups with 10 replicates in each group and 1 pig per replicate. Pigs in groups 1, 2, 3 and 4 were fed a basal diet supplemented with 0, 125, 250 and 500 mg/kg SI, respectively. Five pigs in each group were slaughtered after 60 d of feeding, and the tissue of testis was sampled for the analysis of tissue morphology and gene expression of Cdh1, SCP3, Tnp1 and Vim. The results showed as follows:supplementation of SI (all doses in the study) induced vacuolization in seminiferous tubules, and decreased the number of mature sperm in tubules; the supplementation significantly decreased the expression levels of Cdh1, SCP3 and Tnp1 genes (P<0.01); the expression level of Vim gene was significantly decreased by 250 mg/kg SI (P<0.05). In conclusion, dietary SI at doses of 125 to 500 mg/kg has obvious injury on testis morphology of Xiang pigs, and inhibits expressions of marker genes during spermatogenesis. Dietary SI at a dose of 250 mg/kg inhibits expression of the marker gene of sustentacular cell of testis.
[1] AKINGBEMI B T,BRADEN T D,KEMPPAINEN B W,et al.Exposure to phytoestrogens in the perinatal period affects androgen secretion by testicular leydig cells in the adult rat[J].Endocrinology,2007,148(9):4475-4488.
[2] 周远忠,申旭波,袁国平,等.大豆异黄酮对雄性子代大鼠生殖系统影响[J].中国公共卫生,2008,24(4):450-451.
[3] CHRISTOPHER R C,CELINE Z,JEAN-LOUIS B,et al.Potential detrimental effects of a phytoestrogen-rich diet on male fertility in mice[J].Molecular and Cellular Endocrinology,2010,321:152-160.
[4] 杨科峰,蔡美琴,毛绚霞,等.大豆异黄酮对邻苯二甲酸二丁酯致小鼠遗传毒性的保护作[J].营养学报,2008,30(2):199-202.
[5] DE KRETSER D M,LOVELAND K L,MEINHARDT A,et al.Spermatogenesis[J].Human Reproduction,1998,13(1):1-8.
[6] TOKUDA M,KADOKAWA Y,KURAHASHI H,et al.CDH1 is a specific marker for undifferentiated spermatogonia in mouse testes[J].Biology of Reproduction,2007,76:130-141.
[7] YUAN L,LIU J G,ZHAO J,et al.The murine SCP3 gene is required for synaptonemal complex assembly,chromosome synapsis,and male fertility[J].Molecular Cell,2000,5(1):73-83.
[8] MARRET C,AVALLET O,PERRARD-SAPORI M H,et al.Localization and quantitative expression of mRNAs encoding the testis-specific histone TH2B,the phosphoprotein P19,the transition proteins 1 and 2 during pubertal development and throught the spermatogenic cycle of the rat[J].Molecular Reproductive Development,1998,51:22-35.
[9] PAYNE R L,BINDNER T D,SOUTHERN L L,et al.Effects of dietary soy isoflavones on growth,carcass traits,and meat quality in growing-finishing pigs[J].Journal of Animal Science,2001,79:1230-1239.
[10] 纪冬,辛绍杰.实时荧光定量PCR的发展和数据分析[J].生物技术通讯,2009,20(4):598-600.
[11] YOKO S,SHIARI N,TERUAKI I.Study of spermatogenesis and thickening of lamina propria in the human seminiferous tubules[J].Fertility and Sterility,2008,90(4):1310-1312.
[12] FUBIN W,QIANGGE Z,JINGLI C,et al.The microtubule plus end-binding protein EB1 is involved in Sertoli cell plasticity in testicular seminiferous tubules[J].Experimental Cell Research,2008,314:213-226.
[13] DELCLOS K B,THOMAS J B,LARYY G L,et al.Effects of dietary genistein exposure during development on male and female CD (Sprague-Dawley) rats[J].Reproductive Toxicology,2001,15:647-663.
[14] KATARZYNA P,IRENA B B,MARIOLA M,et al.Changes in male reproductive system and mineral metabolism induced by soy isoflavones administered to rats from prenatal life until sexual maturity[J].Nutrition,2011,27(3):372-279.
[15] PELTTARI J,HOJA M R,YUAN L,et al.Meiotic chromosomal core consisting of cohesion complex proteins recruits DNA recombination proteins and promotes synapsis in the absence of an axial element in mammalian meiotic cells[J].Molecular Cell Biology,2001,21:5667-5677.
[16] YU Y E,ZHANG Y,UNNI E,et al.Abnormal spermatogenesis and reduced fertility in transition nuclear protein 1-deficient mice[J].Proceedings of the National Academy of Sciences,2000,97:4683-4688.
[17] DELLA-MARIA J,GERARD A,FRANCK P,et al.Effects of androgen-binding protein (ABP) on spermatid Tnp1 gene expression in vitro[J].Molecular and Cellular Endocrinology,2002,198:131-141.
[18] SOUAD H M,BEKHEET,RALF S.Disruption of gap junctional intercellular communication by antibiotic gentamicin is associated with aberrant localization of occludin,N-cadherin,connexin 43,and vimentin in SerW3 Sertoli cells in vitro[J].Environmental Toxicology and Pharmacology,2009,28:155-160.
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