聚精颗粒对弱精子症患者顶体酶、核蛋白组型转换、CDY基因和CatSper基因的实验及临床研究
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摘要
男性不育症是男科目前研究的热点和难点,精液异常是男性不育症的主要原因,而其中弱精子症是精液异常的主要类型。导师徐福松教授在此方面积累了丰富的经验,运用导师经验方“聚精颗粒”治疗此类病人效果卓越,但其机理尚不明确,尤其对精子顶体酶和精子核蛋白组型转换两个方面,以及CDY基因和CatSper基因表达方面。本论文共分六个部分,在理论和实验两方面作了总结和研究。
1对中医文献进行系统复习,回顾了男性不育症的历史渊源、发展认识和目前中医对其病因、病机、治疗、研究的概况。尤其是总结了目前对弱精子症患者的中医治疗现状。
2对弱精子症的现代医学的研究情况作了概述,主要总结了弱精子症的细胞因子、DNA、遗传学、蛋白质等方面研究现状。
3对弱精子症的现代治疗现状作了概述,主要总结了激素、左旋肉碱和乙酰左旋肉碱、维生素和辅助生殖等方面的治疗状况。
4对弱精子症患者的精子顶体酶和精子核蛋白组型转换进行了实验研究,并分析了弱精子症患者的精子顶体酶和精子核蛋白组型转换的临床特点和精液参数的关系。与正常生育组相比,299例弱精子症组患者除精子密度外,活力a级精子、活力a+b级精子、精子顶体酶值和核蛋白组型转换半定量值差异有显著性,p<0.01。与正常生育组相比,63例顶体酶异常性弱精子症患者的精子密度(p<0.05)、活力a级精子、活力a+b级精子、精子顶体酶值和核蛋白组型转换半定量值差异有显著性,p<0.01。与正常生育组相比,57例核蛋白组型异常性弱精子症组除精子密度外,活力a级精子、活力a+b级精子、精子顶体酶值和核蛋白组型转换半定量值差异有显著性,p<0.01。结果表明,顶体酶活性和精子核蛋白组型转换分别与精子密度、a级及a+b级活力之间具有相关性,精子顶体酶异常和核蛋白组型转换异常是导致男性不育的主要原因之一。
5运用聚精颗粒对精子顶体酶异常和精子核蛋白组型转换异常的弱精子症患者进行临床治疗,观察到58例治疗后精子顶体酶明显上升,与治疗前比较有显著性差异(p<0.01)。、治疗前后精液各参数比较,除精子密度外(p<0.05),a级精子、(a+b级)精子有显著差异性,p<0.01,精液质量明显改善。而63例核蛋白组型异常组中,精子密度治疗前后有差异性(P<0.05),而治疗后精子核蛋白组型半定量值和a级精子、(a+b级)精子、活率等主要精液参数指标,均有显著改善(p<0.01)。本研究的结果显示,聚精颗粒能促进顶体酶异常性弱精子症患者的精子顶体酶值升高,促进核蛋白组型异常性弱精子症患者的精子核蛋白组型转换半定量值下降,提高生育力和改善精液质量。总有效率分别达81%和80.9%,表明了聚精颗粒对弱精子症患者精子质量良好的调节作用,因此具有确切的有效性和良好的安全性。
6在对精子核蛋白组型转换异常弱精子症患者的CDY基因和CatSper基因表达进行实验研究中,我们观察到,CDY基因检测结果比较完整,CatSper基因表达比较低。聚精颗粒治疗前后的CDY基因表达量分别是治疗前的2.75倍、2.32倍、0.60倍和0.14倍。有2例聚精颗粒治疗前后的CatSper基因表达量是治疗前的0.99倍和0.093倍。我们初步认为,聚精颗粒通过CDY基因表达增强,明确CDY基因具有对精子核蛋白组型转换的调节作用,即CDY基因高表达能够降低核蛋白组型转换值的作用,而且聚精颗粒具有促进CDY基因表达作用,提示CDY基因表达调节作用是至关重要的。而CatSper的基因调节作用尚需进一步研究。
Male infertility is not only the hot topic, but also the therapeutic nodus. Male infertily is main due to dysspermia, asthenospermatism is main type. Professor Xu Fu Song has affluent experience and his empirical formula of "Jujing granulation"is effective in treating this disease, however the mechanism is not clear. Our reseach includes theory exploration and experiment reseach. The article is divided to six parts
1. We reviewed the traditional Chinese medicine(TCM) documents and looked back to the origin and history of male infertility. We also reviewed pathogenesis, therapeutics and reseach in TCM. Especially we summarized the methods have treated for asthenospermia by TCM at present.
2. We summarized the documents of asthenospermia in modern medicine, mainly summarized the study of cytokine, DNA, genetics and protein in asthenospermia.
3. We summarized the therapeutics of asthenospermia in modern medicine, mainly summarized the treatment methods of hormone, L-camitine and L-acetyl-camitine, vitamin, assisted reproductive technology(ART) in asthenospermia.
4. observe the change of spermatozoa acrosin activity and spermatozoa nucleoprotein transition in asthenospermia patient, study the relationship of spermatozoa acrosin activity, spermatozoa nucleoprotein transition and seminal parameter in these patients. Compare with normal breeding group, except sperm concentration, rapid motility, forward motility, acrosin and nucleoprotein transition changed significantly(p<0.01) in 299 cases of asthenospermia patient. For 63 cases of asthenospermatism with abnormal acrosin, compare with normal breeding group, sperm concentration changed(p<0.05), rapid motility, forward motility, acrosin and nucleoprotein transition changed significantly(p<0.01). For 57 cases of asthenospermatism with abnormal nucleoprotein transition, compare with normal breeding group, except sperm concentration, rapid motility, forward motility, acrosin and nucleoprotein transition changed significantly(p<0.01). acrosin and nucleoprotein transition is correlation with sperm concentration, rapid motility and forward motility, Abnormal acrosin and abnormal nucleoprotein transition are one of excellence causes for male infertility.
5. The asthenospermia patients have treated by Jujing granulation, We observed, compare with normal breeding group, sperm concentration changed (p<0.05), rapid motility, forward motility, acrosin and nucleoprotein transition changed significantly(p<0.01) in 58 cases of asthenospermia patient with abnormal acrosin. For 63 cases of asthenospermatism with abnormal nucleoprotein transition, sperm concentration changed (p<0.05), rapid motility, forward motility, acrosin and nucleoprotein transition changed significantly(p<0.01). The Jujing granulation have good curative effect on asthenospermia patient with abnormal acrosin and abnormal nucleoprotein transition. It may improved acrosin, cut down nucleoprotein transition, inproved sperm quality. The Jujing granulation have good curative effect on male sterility.
6. In study of the expressing of CDY and CatSper in 4 cases of asthenospermatism with abnormal nucleoprotein transition, we observed,CDY expressed has completed, CatSper expressed was low.After treated by Jujing granulation, the quantity of CDY expressed were 2.75,2.32,0.60 and 0.14 times respectively compared with pre-treated. The quantity of hCatSper expressed were 0.99 and 0.093 times respectively in only 2 patients. We initianlly think that CDY expressed could accommodate protamines replaced histones, CDY expressed could elevated by Jujing granulation, and cut down nucleoprotein transition. The role of CatSper expressed need more study.
1对中医文献进行系统复习,回顾了男性不育症的历史渊源、发展认识和目前中医对其病因、病机、治疗、研究的概况。尤其是总结了目前对弱精子症患者的中医治疗现状。
2对弱精子症的现代医学的研究情况作了概述,主要总结了弱精子症的细胞因子、DNA、遗传学、蛋白质等方面研究现状。
3对弱精子症的现代治疗现状作了概述,主要总结了激素、左旋肉碱和乙酰左旋肉碱、维生素和辅助生殖等方面的治疗状况。
4对弱精子症患者的精子顶体酶和精子核蛋白组型转换进行了实验研究,并分析了弱精子症患者的精子顶体酶和精子核蛋白组型转换的临床特点和精液参数的关系。与正常生育组相比,299例弱精子症组患者除精子密度外,活力a级精子、活力a+b级精子、精子顶体酶值和核蛋白组型转换半定量值差异有显著性,p<0.01。与正常生育组相比,63例顶体酶异常性弱精子症患者的精子密度(p<0.05)、活力a级精子、活力a+b级精子、精子顶体酶值和核蛋白组型转换半定量值差异有显著性,p<0.01。与正常生育组相比,57例核蛋白组型异常性弱精子症组除精子密度外,活力a级精子、活力a+b级精子、精子顶体酶值和核蛋白组型转换半定量值差异有显著性,p<0.01。结果表明,顶体酶活性和精子核蛋白组型转换分别与精子密度、a级及a+b级活力之间具有相关性,精子顶体酶异常和核蛋白组型转换异常是导致男性不育的主要原因之一。
5运用聚精颗粒对精子顶体酶异常和精子核蛋白组型转换异常的弱精子症患者进行临床治疗,观察到58例治疗后精子顶体酶明显上升,与治疗前比较有显著性差异(p<0.01)。、治疗前后精液各参数比较,除精子密度外(p<0.05),a级精子、(a+b级)精子有显著差异性,p<0.01,精液质量明显改善。而63例核蛋白组型异常组中,精子密度治疗前后有差异性(P<0.05),而治疗后精子核蛋白组型半定量值和a级精子、(a+b级)精子、活率等主要精液参数指标,均有显著改善(p<0.01)。本研究的结果显示,聚精颗粒能促进顶体酶异常性弱精子症患者的精子顶体酶值升高,促进核蛋白组型异常性弱精子症患者的精子核蛋白组型转换半定量值下降,提高生育力和改善精液质量。总有效率分别达81%和80.9%,表明了聚精颗粒对弱精子症患者精子质量良好的调节作用,因此具有确切的有效性和良好的安全性。
6在对精子核蛋白组型转换异常弱精子症患者的CDY基因和CatSper基因表达进行实验研究中,我们观察到,CDY基因检测结果比较完整,CatSper基因表达比较低。聚精颗粒治疗前后的CDY基因表达量分别是治疗前的2.75倍、2.32倍、0.60倍和0.14倍。有2例聚精颗粒治疗前后的CatSper基因表达量是治疗前的0.99倍和0.093倍。我们初步认为,聚精颗粒通过CDY基因表达增强,明确CDY基因具有对精子核蛋白组型转换的调节作用,即CDY基因高表达能够降低核蛋白组型转换值的作用,而且聚精颗粒具有促进CDY基因表达作用,提示CDY基因表达调节作用是至关重要的。而CatSper的基因调节作用尚需进一步研究。
Male infertility is not only the hot topic, but also the therapeutic nodus. Male infertily is main due to dysspermia, asthenospermatism is main type. Professor Xu Fu Song has affluent experience and his empirical formula of "Jujing granulation"is effective in treating this disease, however the mechanism is not clear. Our reseach includes theory exploration and experiment reseach. The article is divided to six parts
1. We reviewed the traditional Chinese medicine(TCM) documents and looked back to the origin and history of male infertility. We also reviewed pathogenesis, therapeutics and reseach in TCM. Especially we summarized the methods have treated for asthenospermia by TCM at present.
2. We summarized the documents of asthenospermia in modern medicine, mainly summarized the study of cytokine, DNA, genetics and protein in asthenospermia.
3. We summarized the therapeutics of asthenospermia in modern medicine, mainly summarized the treatment methods of hormone, L-camitine and L-acetyl-camitine, vitamin, assisted reproductive technology(ART) in asthenospermia.
4. observe the change of spermatozoa acrosin activity and spermatozoa nucleoprotein transition in asthenospermia patient, study the relationship of spermatozoa acrosin activity, spermatozoa nucleoprotein transition and seminal parameter in these patients. Compare with normal breeding group, except sperm concentration, rapid motility, forward motility, acrosin and nucleoprotein transition changed significantly(p<0.01) in 299 cases of asthenospermia patient. For 63 cases of asthenospermatism with abnormal acrosin, compare with normal breeding group, sperm concentration changed(p<0.05), rapid motility, forward motility, acrosin and nucleoprotein transition changed significantly(p<0.01). For 57 cases of asthenospermatism with abnormal nucleoprotein transition, compare with normal breeding group, except sperm concentration, rapid motility, forward motility, acrosin and nucleoprotein transition changed significantly(p<0.01). acrosin and nucleoprotein transition is correlation with sperm concentration, rapid motility and forward motility, Abnormal acrosin and abnormal nucleoprotein transition are one of excellence causes for male infertility.
5. The asthenospermia patients have treated by Jujing granulation, We observed, compare with normal breeding group, sperm concentration changed (p<0.05), rapid motility, forward motility, acrosin and nucleoprotein transition changed significantly(p<0.01) in 58 cases of asthenospermia patient with abnormal acrosin. For 63 cases of asthenospermatism with abnormal nucleoprotein transition, sperm concentration changed (p<0.05), rapid motility, forward motility, acrosin and nucleoprotein transition changed significantly(p<0.01). The Jujing granulation have good curative effect on asthenospermia patient with abnormal acrosin and abnormal nucleoprotein transition. It may improved acrosin, cut down nucleoprotein transition, inproved sperm quality. The Jujing granulation have good curative effect on male sterility.
6. In study of the expressing of CDY and CatSper in 4 cases of asthenospermatism with abnormal nucleoprotein transition, we observed,CDY expressed has completed, CatSper expressed was low.After treated by Jujing granulation, the quantity of CDY expressed were 2.75,2.32,0.60 and 0.14 times respectively compared with pre-treated. The quantity of hCatSper expressed were 0.99 and 0.093 times respectively in only 2 patients. We initianlly think that CDY expressed could accommodate protamines replaced histones, CDY expressed could elevated by Jujing granulation, and cut down nucleoprotein transition. The role of CatSper expressed need more study.
引文
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