小分子牡蛎多肽对雄性小鼠性功能的影响
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摘要
探究不同剂量的小分子牡蛎多肽对雄性小鼠性功能的影响。将6周龄的雄性昆明小鼠随机分成4组,连续灌胃6周。分别测定每组小鼠交配能力,性腺器官的重量和脏器指数,精子数目、活力和畸形率;采用分光光度法测定小鼠血清中胆固醇和阴茎中NO含量;采用ELISA法测定小鼠血清中睾酮(testosterone, T)、黄体生成素(luteinizing hormone, LH)和卵泡刺激素(follicle stimulating hormone, FSH)的含量;采用实时荧光定量PCR检测SRPK2及3个雄激素限速酶(类固醇生成急性调节蛋白(StAR)、细胞色素P450胆固醇侧链裂解酶(P450scc)和3β-羟基类固醇脱氢酶(3β-HSD))基因的表达情况。结果表明,小分子牡蛎多肽可以通过提高小鼠的交配能力、性腺器官重量和脏器系数以及精子质量,增加小鼠血清中胆固醇和性激素以及阴茎中NO的含量,提高性功能相关基因的表达量来增强雄性小鼠的性功能。在一定剂量范围内,性功能的增强程度具有一定剂量依赖性。
To explore the effects of different doses of small molecule polypeptide of oyster on sexual function in male mice, male mice of 6-week-old were randomly divided into 4 groups and continued gavage for 6 weeks. The mating ability, weight and organ coefficient of gonad, sperm count, motility and malformation rate of mice in each group were determined respectively. The levels of cholesterol in serum and NO in penis of mice were tested by spectrophotometry, as well as the contents of testosterone(T), luteinizing hormone(LH) and follicle stimulating hormone(FSH) in serum of mice were determined by ELISA method. The expression of SRPK2 and three androgen rate-limiting enzymes genes(StAR, P450 scc, and 3β-HSD) were detected by using RT-PCR. The results showed that small molecule polypeptide of oyster could enhance sexual function of male mice by improving the mating ability, weight and organ coefficient of gonad and sperm qualities, increasing the content of cholesterol and sex hormones in serum and NO in penis, and improving the expression of sexual function related genes in mice.Moreover, the degree of sexual function enhancement could have a dose-dependent in a certain range of dose.
To explore the effects of different doses of small molecule polypeptide of oyster on sexual function in male mice, male mice of 6-week-old were randomly divided into 4 groups and continued gavage for 6 weeks. The mating ability, weight and organ coefficient of gonad, sperm count, motility and malformation rate of mice in each group were determined respectively. The levels of cholesterol in serum and NO in penis of mice were tested by spectrophotometry, as well as the contents of testosterone(T), luteinizing hormone(LH) and follicle stimulating hormone(FSH) in serum of mice were determined by ELISA method. The expression of SRPK2 and three androgen rate-limiting enzymes genes(StAR, P450 scc, and 3β-HSD) were detected by using RT-PCR. The results showed that small molecule polypeptide of oyster could enhance sexual function of male mice by improving the mating ability, weight and organ coefficient of gonad and sperm qualities, increasing the content of cholesterol and sex hormones in serum and NO in penis, and improving the expression of sexual function related genes in mice.Moreover, the degree of sexual function enhancement could have a dose-dependent in a certain range of dose.
引文
Canseco-Alba A.,and Rodríguez-Manzo G.,2016,Intra-VTAanandam ide infusion produces dose-based biphasic effects on male rat sexual behavior expression,Pharmacology,Biochemistry and Behavior,150-115:182-189
Bao L.,Wang J.B.,Zhang Y.,Cai X.X.,Sun B.,Chen Q.H.,Ren J.W.,Zhang Z.F.,and Li Y.,2015,Effects of panax ginseng oligopeptide of Jilin on sexual function in male mice,Zhongguo Yufang Yixue Zazhi(China Preventive Medicine),16(10):757-760(鲍雷,王军波,张远,蔡夏夏,孙彬,陈启贺,任金威,张召锋,李勇,2015,吉林人参低聚肽对雄性小鼠性功能影响的实验研究,中国预防医学杂志,16(10):757-760)
Cui J.M.,Wang X.Y.,Liu H.Y.,Zhang L.,and Song X.X.,2013,Effects of Xuefurong capsules on sexual function and nNOSexpression in paraventricular nucleus in male castrated rats,Xi'an Jiaotong Daxue Xuebao(Journal of Xi'an Jiaotong U-niversity(Medical Sciences)),31(5):588-591(崔建梅,汪晓阳,刘鸿宇,张磊,宋晓霞,2013,雪芙蓉胶囊对去势雄性大鼠性功能和下丘脑室旁核一氧化氮合酶表达的影响,西安交通大学学报(医学版),31(5):588-591)
Feng Q.L.,2016,Method for extracting polypeptide of small molecule oyster,China Patent,CN201610894500.3(冯群力,2016,一种小分子牡蛎多肽的提取方法,中国专利,CN201610894500.3)
Lagoda G.,Muschamp J.W.,Vigdorchik A.,and Hull E.M.,2004,A nitric oxide synthesis inhibitor in the medial preoptic area inhibits copulation and stimulus sensitization in male rats,Behav.Neurosci.,118(6):1317-1323
Lin M.D.,Chen Y.,Chen T.H.,and Chen H.,2008,Effects of low power microwave on serum testosterone and cholesterol level in male mice,Zhongguo Gonggong Weisheng(Chinese Journal of Public Health),24(10):1206-1207(林孟端,陈昱,陈铁晖,陈华,2008,低功率微波辐射对雄性小鼠性功能影响,中国公共卫生,24(10):1206-1207)
Liu M.H.,Huang Q.L.,Liu Z.F.,and Shui P.X.,2016,Effect of penthorum Chinense pursh on sexual function of male rats,Luzhou Yixueyuan Xuebao(Journal of Luzhou Medical College),39(6):547-550(刘明华,黄群莲,刘中飞,税丕先,2016,赶黄草对雄性大鼠正常性功能的影响,泸州医学院学报,39(6):547-550)
Lin Y.Y.,Li K.C.,Du L.,Miao X.,Lang H.Y.,Liu J.Y.,Zeng L.H.,and Guo G.Z.,2014,Effect of 1 950 MHz radio frequency electromagnetic radiation on testosterone secretion in mice testicular leydig cells,Gaoyadian Jishu(High Voltage Engineering),40(12):3854-3859(林艳云,李康樗,杜乐,苗霞,郎海洋,刘军叶,曾丽华,郭国祯,2014,1 950 MHz射频电磁辐射对小鼠睾丸间质细胞睾酮分泌的影响,高压电技术,40(12):3854-3859)
Luo Q.,Huang X.L.,Li Z.N.,Yang M.L.,and Yan J.,2006,Effect of lycium barbarum polysaccharides on sexual function and reproductive function of male rats,Yingyang Xuebao(Acta Nutrimenta Sinica),28(1):62-70(罗琼,黄晓兰,李卓能,杨明亮,阎俊,2006,枸杞多糖对雄性大鼠性功能及生殖功能的影响,营养学报,28(1):62-70)
Luo Q.,Ren S.C.,Yang M.L.,Yan J.,and Yang J.J.,2009,Effect of laminarina japonica polysacchocrideson sexual function of male rats after irradiation of testis,Yingyang Xuebao(Acta Nutrimenta Sinica),31(3):259-262(罗琼,任世成,杨明亮,闫俊,杨佳娟,2009,海带多糖对睾丸受辐射雄性大鼠性功能的影响,营养学报,31(3):259-262)
Luo Q.J.,Lu S.B.,Li H.,Jing Z.,Xiong J.Y.,and Zhang Y.J.,2013,The Effect of oyster peptides supplementation on serum testosterone,LH content and St AR m RNA expression in rats during the long-term and high loading training,Jiangxi Shifan Daxue Xuebao(Journal of Jiangxi Normal U-niversity(Natural Science Edition)),37(6):611-616(罗齐军,鲁顺保,李红,靖洲,熊静宇,张艳杰,2013,牡蛎多肽对长期大负荷训练大鼠血睾酮、LH和St AR m RNA表达的影响,江西师范大学学报(自然科学版),37(6):611-616)
Lu X.M.,Wang Q.L.,Lang X.Y.,Chen J.S.,Yang Y.C.,and Zhang L.,2013,Study on the antimicrobial and antioxidant activities of bioactive peptides from oyster,Zhongguo Niangzao(China Brewing),32(2):77-79(卢学敏,王颀林,蓝晓燕,陈俊生,杨宇晨,张莉,2013,牡蛎活性多肽的抑菌作用与抗氧化性能研究,中国酿造,32(2):77-79)
Retana-Márquez S.,Bonilla-Jaime H.,Vázquez-Palacios G.,Martínez-García R.,and Velázquez-Moctezuma J.,2003,Changes in masculine sexual behavior,corticosterone and testosterone in response to acute and chronic stress in male rats,Horm Behav.,44(4):327-337
Wang B.X.,Wang S.Q.,Wang S.X.,and Gao Y.H.,2004,Study of the effect of super painkiller on the sexual function of male rats with Type l diabetes,Zhonghua Nankexue Zazhi(National Journal of Andrology),10(7):521-523(王柏欣,王淑秋,王淑湘,高艳华,2004,镇痛灵对雄性l型糖尿病大鼠性功能的影响,中华男科学杂志,10(7):521-523)
Wang J.R.,Wang H.W.,Xu X.,Xu S.H.,and Xue L.X.,2002,Effect of opuntia extracts on sex function in male mice,Haixia Yaoxue(Strait Pharmaceutical Journal),14(1):16-17(王建人,王红伟,徐霞,许世华,薛乐勋,2002,仙人掌提取物对雄性小鼠性功能的影响,海峡药学,14(1):16-17)
Wang X.Y.,and Zhang J.T.,2000,Effect of ginsenoside Rb1 on mouse sexual function and its mechanism,Yaoxue Xuebao(Acta Pharmaceutica Sinica),35(7):492-495(王晓英,张均田,2000,人参皂苷Rb1对小鼠性功能的改善作用及其机制探讨,药学学报,35(7):492-495)
Xu K.S.,Jiang H.,Hang Y.Q.,Xue H.Q.,and Xia D.,2010,The effect of endocrine disruptor fenvalerate on thereproductive physiology of adult male mice,Shanghai Nongye Xuebao(Acta Agriculturae Shanghai),26(2):5-8(徐克斯,蒋辉,杭怡琼,薛惠琴,夏东,2010,内分泌干扰素氰戊菊酯对成年雄性小鼠繁殖生理的影响,上海农业学报,26(2):5-8)
Yang H.,Li X.B.,Li T.,Jia S.S.,Wang H.L.,and Song G.H.,2015,Expression and significance of SR-protein-specific kinase SRPK2 in mouse testis,Zhongguo Shiyan Dongwu Xuebao(Acta Laboratorium Animalis Scientia Sinica),23(2):171-176(杨红,李晓宾,李婷,贾三三,王海龙,宋国华,郭睿,2015,丝氨酸/精氨酸蛋白特异激酶SRPK2在小鼠睾丸组织中的表达及意义,中国实验动物学报,23(2):171-176)
Yu J.,Yang Z.G.,Chen M.Z.,Zhong L.,and Li W.,2012,Induction and mechanism of heLa cell apoptosis by bioactive peptide from oyster,Shipin Kexue(Food Science),33(21):290-294(余杰,杨振国,陈美珍,钟炼,李伟,2012,牡蛎活性肽体外诱导HeLa细胞凋亡及其作用机制,食品科学,33(21):290-294)
Zang Z.J.,Tang H.F.,Tuo Y.,Xing W.J.,Ji S.Y.,Gao Y.,and Deng C.H.,2016,Effects of velvet antler polypeptide on sexual behavior and testosterone synthesis in aging male mice,Asian J.Androl.,18(4):613-619
Zhang J.,and Chen L.Y.,2012,Study on effect of black fungus poIysaccharide on sexual function of adolescence male rats,Xiandai Zhongxiyi Jiehe Zazhi(Modern Journal of Integrated Traditional Chinese and Western Medicine),21(33):3671-3672(张洁,陈立勇,2012,黑木耳多糖对青春期雄性大鼠性功能影响的研究,现代中西医结合杂志,21(33):3671-3672)
Zheng X.,2013,Lamotrigine combining with valproic acid on sex hormones and sexual function in male epilepsy patients,Zhongguo Xingkexue(The Chinese Journal of Human Sexuality),22(9):6-9(郑霞,2013,拉莫三嗪联合丙戊酸对男性癫痫患者性激素水平、性功能变化的影响,中国性科学,22(9):6-9)
Zhou D.X.,Qiu S.D.,Wang Z.Y.,and Zhang J.,2006,Effect of tail-suspension on the reproduction of adult male rats,Zhonghua Nankexue Zazhi(National Journal of Andrology),12(4):326-329(周党侠,邱曙东,王志勇,张洁,2006,尾悬吊状态对性成熟期雄性大鼠生殖功能的影响,中华男科学杂志,12(4):326-329)
Bao L.,Wang J.B.,Zhang Y.,Cai X.X.,Sun B.,Chen Q.H.,Ren J.W.,Zhang Z.F.,and Li Y.,2015,Effects of panax ginseng oligopeptide of Jilin on sexual function in male mice,Zhongguo Yufang Yixue Zazhi(China Preventive Medicine),16(10):757-760(鲍雷,王军波,张远,蔡夏夏,孙彬,陈启贺,任金威,张召锋,李勇,2015,吉林人参低聚肽对雄性小鼠性功能影响的实验研究,中国预防医学杂志,16(10):757-760)
Cui J.M.,Wang X.Y.,Liu H.Y.,Zhang L.,and Song X.X.,2013,Effects of Xuefurong capsules on sexual function and nNOSexpression in paraventricular nucleus in male castrated rats,Xi'an Jiaotong Daxue Xuebao(Journal of Xi'an Jiaotong U-niversity(Medical Sciences)),31(5):588-591(崔建梅,汪晓阳,刘鸿宇,张磊,宋晓霞,2013,雪芙蓉胶囊对去势雄性大鼠性功能和下丘脑室旁核一氧化氮合酶表达的影响,西安交通大学学报(医学版),31(5):588-591)
Feng Q.L.,2016,Method for extracting polypeptide of small molecule oyster,China Patent,CN201610894500.3(冯群力,2016,一种小分子牡蛎多肽的提取方法,中国专利,CN201610894500.3)
Lagoda G.,Muschamp J.W.,Vigdorchik A.,and Hull E.M.,2004,A nitric oxide synthesis inhibitor in the medial preoptic area inhibits copulation and stimulus sensitization in male rats,Behav.Neurosci.,118(6):1317-1323
Lin M.D.,Chen Y.,Chen T.H.,and Chen H.,2008,Effects of low power microwave on serum testosterone and cholesterol level in male mice,Zhongguo Gonggong Weisheng(Chinese Journal of Public Health),24(10):1206-1207(林孟端,陈昱,陈铁晖,陈华,2008,低功率微波辐射对雄性小鼠性功能影响,中国公共卫生,24(10):1206-1207)
Liu M.H.,Huang Q.L.,Liu Z.F.,and Shui P.X.,2016,Effect of penthorum Chinense pursh on sexual function of male rats,Luzhou Yixueyuan Xuebao(Journal of Luzhou Medical College),39(6):547-550(刘明华,黄群莲,刘中飞,税丕先,2016,赶黄草对雄性大鼠正常性功能的影响,泸州医学院学报,39(6):547-550)
Lin Y.Y.,Li K.C.,Du L.,Miao X.,Lang H.Y.,Liu J.Y.,Zeng L.H.,and Guo G.Z.,2014,Effect of 1 950 MHz radio frequency electromagnetic radiation on testosterone secretion in mice testicular leydig cells,Gaoyadian Jishu(High Voltage Engineering),40(12):3854-3859(林艳云,李康樗,杜乐,苗霞,郎海洋,刘军叶,曾丽华,郭国祯,2014,1 950 MHz射频电磁辐射对小鼠睾丸间质细胞睾酮分泌的影响,高压电技术,40(12):3854-3859)
Luo Q.,Huang X.L.,Li Z.N.,Yang M.L.,and Yan J.,2006,Effect of lycium barbarum polysaccharides on sexual function and reproductive function of male rats,Yingyang Xuebao(Acta Nutrimenta Sinica),28(1):62-70(罗琼,黄晓兰,李卓能,杨明亮,阎俊,2006,枸杞多糖对雄性大鼠性功能及生殖功能的影响,营养学报,28(1):62-70)
Luo Q.,Ren S.C.,Yang M.L.,Yan J.,and Yang J.J.,2009,Effect of laminarina japonica polysacchocrideson sexual function of male rats after irradiation of testis,Yingyang Xuebao(Acta Nutrimenta Sinica),31(3):259-262(罗琼,任世成,杨明亮,闫俊,杨佳娟,2009,海带多糖对睾丸受辐射雄性大鼠性功能的影响,营养学报,31(3):259-262)
Luo Q.J.,Lu S.B.,Li H.,Jing Z.,Xiong J.Y.,and Zhang Y.J.,2013,The Effect of oyster peptides supplementation on serum testosterone,LH content and St AR m RNA expression in rats during the long-term and high loading training,Jiangxi Shifan Daxue Xuebao(Journal of Jiangxi Normal U-niversity(Natural Science Edition)),37(6):611-616(罗齐军,鲁顺保,李红,靖洲,熊静宇,张艳杰,2013,牡蛎多肽对长期大负荷训练大鼠血睾酮、LH和St AR m RNA表达的影响,江西师范大学学报(自然科学版),37(6):611-616)
Lu X.M.,Wang Q.L.,Lang X.Y.,Chen J.S.,Yang Y.C.,and Zhang L.,2013,Study on the antimicrobial and antioxidant activities of bioactive peptides from oyster,Zhongguo Niangzao(China Brewing),32(2):77-79(卢学敏,王颀林,蓝晓燕,陈俊生,杨宇晨,张莉,2013,牡蛎活性多肽的抑菌作用与抗氧化性能研究,中国酿造,32(2):77-79)
Retana-Márquez S.,Bonilla-Jaime H.,Vázquez-Palacios G.,Martínez-García R.,and Velázquez-Moctezuma J.,2003,Changes in masculine sexual behavior,corticosterone and testosterone in response to acute and chronic stress in male rats,Horm Behav.,44(4):327-337
Wang B.X.,Wang S.Q.,Wang S.X.,and Gao Y.H.,2004,Study of the effect of super painkiller on the sexual function of male rats with Type l diabetes,Zhonghua Nankexue Zazhi(National Journal of Andrology),10(7):521-523(王柏欣,王淑秋,王淑湘,高艳华,2004,镇痛灵对雄性l型糖尿病大鼠性功能的影响,中华男科学杂志,10(7):521-523)
Wang J.R.,Wang H.W.,Xu X.,Xu S.H.,and Xue L.X.,2002,Effect of opuntia extracts on sex function in male mice,Haixia Yaoxue(Strait Pharmaceutical Journal),14(1):16-17(王建人,王红伟,徐霞,许世华,薛乐勋,2002,仙人掌提取物对雄性小鼠性功能的影响,海峡药学,14(1):16-17)
Wang X.Y.,and Zhang J.T.,2000,Effect of ginsenoside Rb1 on mouse sexual function and its mechanism,Yaoxue Xuebao(Acta Pharmaceutica Sinica),35(7):492-495(王晓英,张均田,2000,人参皂苷Rb1对小鼠性功能的改善作用及其机制探讨,药学学报,35(7):492-495)
Xu K.S.,Jiang H.,Hang Y.Q.,Xue H.Q.,and Xia D.,2010,The effect of endocrine disruptor fenvalerate on thereproductive physiology of adult male mice,Shanghai Nongye Xuebao(Acta Agriculturae Shanghai),26(2):5-8(徐克斯,蒋辉,杭怡琼,薛惠琴,夏东,2010,内分泌干扰素氰戊菊酯对成年雄性小鼠繁殖生理的影响,上海农业学报,26(2):5-8)
Yang H.,Li X.B.,Li T.,Jia S.S.,Wang H.L.,and Song G.H.,2015,Expression and significance of SR-protein-specific kinase SRPK2 in mouse testis,Zhongguo Shiyan Dongwu Xuebao(Acta Laboratorium Animalis Scientia Sinica),23(2):171-176(杨红,李晓宾,李婷,贾三三,王海龙,宋国华,郭睿,2015,丝氨酸/精氨酸蛋白特异激酶SRPK2在小鼠睾丸组织中的表达及意义,中国实验动物学报,23(2):171-176)
Yu J.,Yang Z.G.,Chen M.Z.,Zhong L.,and Li W.,2012,Induction and mechanism of heLa cell apoptosis by bioactive peptide from oyster,Shipin Kexue(Food Science),33(21):290-294(余杰,杨振国,陈美珍,钟炼,李伟,2012,牡蛎活性肽体外诱导HeLa细胞凋亡及其作用机制,食品科学,33(21):290-294)
Zang Z.J.,Tang H.F.,Tuo Y.,Xing W.J.,Ji S.Y.,Gao Y.,and Deng C.H.,2016,Effects of velvet antler polypeptide on sexual behavior and testosterone synthesis in aging male mice,Asian J.Androl.,18(4):613-619
Zhang J.,and Chen L.Y.,2012,Study on effect of black fungus poIysaccharide on sexual function of adolescence male rats,Xiandai Zhongxiyi Jiehe Zazhi(Modern Journal of Integrated Traditional Chinese and Western Medicine),21(33):3671-3672(张洁,陈立勇,2012,黑木耳多糖对青春期雄性大鼠性功能影响的研究,现代中西医结合杂志,21(33):3671-3672)
Zheng X.,2013,Lamotrigine combining with valproic acid on sex hormones and sexual function in male epilepsy patients,Zhongguo Xingkexue(The Chinese Journal of Human Sexuality),22(9):6-9(郑霞,2013,拉莫三嗪联合丙戊酸对男性癫痫患者性激素水平、性功能变化的影响,中国性科学,22(9):6-9)
Zhou D.X.,Qiu S.D.,Wang Z.Y.,and Zhang J.,2006,Effect of tail-suspension on the reproduction of adult male rats,Zhonghua Nankexue Zazhi(National Journal of Andrology),12(4):326-329(周党侠,邱曙东,王志勇,张洁,2006,尾悬吊状态对性成熟期雄性大鼠生殖功能的影响,中华男科学杂志,12(4):326-329)