日粮纳米锌水平对公羊睾丸和附睾Cu-ZnSOD表达的影响

doi:10.11843/j.issn.0366-6964.2014.10.007

摘要/Abstract

摘要：

旨在研究日粮纳米锌对公羊睾丸和附睾铜锌超氧化物歧化酶基因（Cu-ZnSOD）mRNA和蛋白表达的影响。将16只9月龄体重相近、健康状况良好的晋中绵羊公羊随机分成4组，分别喂给基础日粮（对照组），基础日粮+纳米锌（50、100和150 mg•kg^-1DM）。预饲期10 d，正试期80 d。试验结束时采集绵羊睾丸和附睾组织。采用RT-PCR、Western blotting和免疫组化技术检测其Cu-ZnSOD mRNA、Cu-ZnSOD蛋白表达及在睾丸中定位情况。结果显示：绵羊睾丸和附睾组织Cu-ZnSOD mRNA表达量顺序是附睾头≥睾丸＞附睾体＞附睾尾。日粮添加50~100 mg•kg^-1纳米锌可增加其睾丸和附睾组织Cu-ZnSOD mRNA和蛋白表达量。免疫组化结果显示Cu-ZnSOD在睾丸的生精细胞中有强阳性信号，间质细胞有弱阳性信号。综上表明，绵羊睾丸和附睾不同区段Cu-ZnSOD mRNA和蛋白表达有差异。日粮添加适量锌可提高睾丸和附睾中Cu-ZnSOD mRNA和蛋白表达量，其作用机制仍需进一步研究。

Abstract:

The objectives of this study were to evaluate the expression of copper zinc superoxide dismutase and its cellular localization in testis and epididymis of ram lambs from different levels of Nano-zinc supplementation groups.Sixteen 9-month-old Jinzhong ram lambs with approximately weight and good healthy were randomly divided into 4 treatments，fed with a basal diet with supplementation of 0，50，100 and 150 mg•kg^-1 DM Nano-zinc respectively.Experimental period was 90 d.At the end of test，the samples of testis and epididymis were collected.Expression of Cu-ZnSOD mRNA and protein was detected by real-time fluorescent quantitative PCR and Western blotting.Cellular localization of Cu-ZnSOD in testis was examined by immunohistochemistry.The quantitative real-time PCR results showed the rank order of Cu-ZnSOD mRNA expression was caput ≥testis > corpora >cauda.The mRNA and protein expression of Cu-ZnSOD in testis and epididymis were higher in the group with supplementation of 50 and 100 mg•kg^-1 Nano-zinc than those in control group（P<0.05）.The strong positive signals of Cu-ZnSOD protein were detected in spermatogenic cells of testis，and weak signals in interstitial tissue.The expression of Cu-ZnSOD was different among testis，caput，corpora and cauda of epididymidis.The optimal supplementation of Nano-zinc in diet could improve the expression of Cu-ZnSOD in testis and epididymis of rams.And the mechanisms needs to be further researched.

中图分类号:

S826.2

任有蛇，秦小伟，郭丽娜，赵辉，刘文忠，张建新，张春香. 日粮纳米锌水平对公羊睾丸和附睾Cu-ZnSOD表达的影响[J]. 畜牧兽医学报, 2014, 45(10): 1622-1630.

REN You-she，QIN Xiao-wei，GUO Li-na，ZHAO Hui，LIU Wen-zhong，ZHANG Jian-xin,ZHANG Chun-xiang. Effect of the Supplementation of Different Nano-zinc Levels on Expression of Copper Zinc Superoxide in Testis and Epididymis of Ram Lambs[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(10): 1622-1630.

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https://www.xmsyxb.com/CN/Y2014/V45/I10/1622

参考文献

［1］BEDWAL R S，NAIR N，MATHUR R S.Effects of zinc deficiency and toxicity on reproductive organs，pregnancy and lactation-a review［J］.Trace Elem Med，1991，8:89-100.
［2］BEDWAL R S，BAHUGUNA A.Zinc，copper and selenium in reproduction［J］.Experientia，1994，50:626-640.
［3］AULD D S.The ins and outs of biological zinc sites［J］. Biometals，2009，22:141-148.
［4］KUMARI D，NAIR N，BEWAL R S.Effect of dietary zinc deficiency on testes of Wistar rats：Morphometric and cell quantification studies［J］.J Trace Elem Med Biol，2011，25(1):47-53.
［5］KUMARI D，NAIR N，BEWAL R S.Testicular apoptosis after dietary zinc deficiency：ultrastructural and TUNEL studies［J］.Syst Biol Reprod Med，2011,57(5):233-243.
［6］WALLACE E，CALVIN H I，SALGO M P，et al.Normal levels of zinc and sulphydryls in morphologically abnormal populations of spermatozoa from moderately zinc deficient rats［J］.Gamete Res，1984，9:375-386.
［7］MERRELLS K J，BLEWETT H，JAMIESON J A，et al.Relationship between abnormal sperm morphology induced by dietary zinc deficiency and lipid composition in testes of growing rats［J］.Br J Nutr， 2009,102(2):226-232.
［8］WANG C L,WU C J，WANG Y R，et al.Study progress of Cu/Zn superoxide disnmtase–from gene to function ［J］.Progr Modern Biomed，2008，8(5)：940-943.
［9］AlVAREZ J G，STOREY B T.Evidence for increased lipid peroxidative damage and loss of superoxide dismutase activity as a model of sublethal cryodamage to human sperm during cryopreservation［J］.J Androl，1992，13:232-242.
［10］SANOCKA D，MIESEL R，JEDRZEJCZAK P，et al.Effect of reactive oxygen species and the activity of antioxidant systems in human semen；association with male infertility［J］.Intern J Andro，1997，20:255-264.
［11］PEEKER R，ABRAMSSON L，MARKLUND S L.Superoxide dismutase isoenzymes in human seminal plasma and spermatozoa［J］. Mol Hum Reprod，1997,3(12):1061-1066.
［12］MRUK D D，SILVESTRINI B，MO M Y，et al.Antioxidant superoxide dismutase–a review：its function，regulation in the testis，and role in male fertility［J］.Contraception，2002,65:305-311.
［13］MURAWSKI M，SACZKO J，MARCINKOWSKA A，et al.Evaluation of superoxide dismutase activity and its impact on semen quality parameters of infertile men［J］.Folia Histochem Cytobiol，2007，45 （Suppl 1）:S123-126.
［14］WAHEED M M，GOUDA E M，KHALIFA T A.Impact of seminal plasma superoxide dismutase and glutathione peroxidase on cryopreserved buffalo spermatozoa［J］.Anim Reprod Sci，2013,142(3-4):126-130.
［15］SØRENSEN M B，STOLTENBERG M，HENRIKSEN K，et al.Histochemical tracing of zinc ions in the rat testis［J］.Mol Hum Reprod，1998,4:423-428.
［16］STOLTENBERG M，SØRENSEN M B，DANSCHER G，et al.Autometallographic demonstration of zinc ions in rat sperm cells［J］.Mol Hum Reprod，1997，3(9):763-767.
［17］FUJIMORI O，TSUKISE A，YAMADA K.Histochemical demonstration of zinc in rat epididymis using a sulphide-silver method［J］.Histochemistry，1988，88(3-6):469-473.
［18］STOLTENBERG M，ERNST E，ANDREASEN A，et al.Histochemical localization of zinc ions in the epididymis of the rat［J］.Histochem J，1996，28(3):173-185.
［19］STOLTENBERG M,LUND L,JUHL S,et al.Histochemical demonstration of zinc ions in human epididymis using autometallography［J］.Histochem J，1997，29(10):721-726.
［20］ROWE M P，POWELL J G，KEGLEY E B，et al.Effect of supplemental trace-mineral source on bull semen quality［J］. Prof Anim Sci，2014,30，1:68-73.
［21］KUMAR N，VERMA R P，SINGH L P，et al.Effect of different levels and sources of zinc supplementation on quantitative and qualitative semen attributes and serum testosterone level in crossbred cattle (Bos indicus ×Bos taurus) bulls［J］.Reprod Nutr Dev，2006，46(6):663-675.
［22］KENDALL N R，MCMULLEN S，GREEN A，et al.The effect of a zinc，cobalt and selenium soluble glass bolus on trace element status and semen quality of ram lambs［J］.Anim Reprod Sci，2000,62(4):277-283.
［23］SAID T M，AGARWAL A，SHARMA R K，et al.Human sperm superoxide anion generation and correlation with semen quality in patients with male infertility［J］.Fertil Steril，2004，82:871-877.
［24］MARZEC-WRÓBLEWSKA U，KAMI?SKI P，LAKOTA 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］KASIMANICKAM R，PELZER K D，KASIMANICKAM V，et al.Association of classical semen parameters，sperm DNA fragmentation index，lipid peroxidation and antioxidant enzymatic activity of semen in ram-lambs［J］.Theriogenology，2006，65(7):1407-1421.
［26］KASIMANICKAM R，KASIMANICKAM V，THATCHER C D，et al.Relationships among lipid peroxidation，glutathione peroxidase，superoxide dismutase，sperm parameters，and competitive index in dairy bulls［J］.Theriogenology，2007，67(5):1004-1012.
［27］CASSANI P，BECONI M T，O'FLAHERTY C.Relationship between total superoxide dismutase activity with lipid peroxidation，dynamics and morphological parameters in canine semen［J］.Anim Reprod Sci，2005，86(1-2):163-173.
［28］ROCA J，RODRIGUEZ M J，GIL M A，et al.Survival and in vitro fertility of boar spermatozoa frozen in the presence of superoxide dismutase and/or catalase［J］. J Andro，2005，26:15-24.
［29］GARRATT M，BATHGATE R，GRAAF S P.Copper-zinc superoxide dismutase deficiency impairs sperm motility and in vivo fertility［J］.Reproduction，2013，146(4):297-304.
［30］TSUNODA S，KAWANO N，MIYADO K，et al.Impaired fertilizing ability of superoxide dismutase 1-deficient mouse sperm during in vitro fertilization［J］.Biol Reprod，2012，87(5):121.
［31］KOBAYASHI T，MIYAZAKI T，NATORI M，et al.Protective role of superoxide dismutase in human sperm motility：superoxide dismutase activity and lipid peroxide in human seminal plasma and spermatozoa［J］.Hum Reprod，1991，6(7):987-991.
［32］CELINO F T，YAMAGUCHI S，MIURA C，et al.Tolerance of spermatogonia to oxidative stress is due to high levels of Zn and Cu/Zn superoxide dismutase［J］.PLoS ONE，2011，6(2):e16938.
［33］BAUMBER J,BALL B A.Determination of glutathione peroxidase and superoxide dismutase-like activities in equine spermatozoa，seminal plasma，and reproductive tissues［J］.Am J Vet Res，2005，66(8):1415-1419.
［34］KOZIOROWSKA-GILUN M，GILUN P，FRASER L，et al.Antioxidant enzyme activity and mRNA expression in reproductive tract of adult male European Bison (Bison bonasus，Linnaeus 1758) ［J］.Reprod Domest Anim，2013，48(1):7-14.
［35］NONOGAKI T，NODA Y，NARIMOTO K，et.al.Localization of CuZn-superoxide dismutase in the human male genital organs ［J］.Hum Reprod，1992，7(1):81-85.
［36］JOW W W，SCHLEGEL P N，CICHON Z，et al.Identification and localization of copper–zinc superoxide dismutase gene expression in rat testicular development［J］.Andrology，1993，14：439-447.

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