针灸对腺嘌呤诱导的少弱精症大鼠生精功能的影响
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摘要
少精子症(oligospermia)是精液中精子的数量低于正常健康有生育能力的男子,是一种较常见的男性不育的病症。现在认为每毫升精液中精子数目低于2000万为少精子症;弱精子症(asthenospermia)是指精液参数中前向运动的精子(a和b级)小于50%或a级运动的精子小于25%的病症。弱精子症又称为精子活力低下。临床上少精子症状常常与精子活率低下同时存在,此时称之为少弱精子症。本病的病因病机非常复杂,遗传因素、内分泌因素、免疫因素、精索静脉曲张、感染因素、放射线、化学品接触等均对精子的生成和成熟有一定程度的影响而造成男性不育。本文总结了历代中医学家对男性不育症的病因、病机、辨证施治的认识,综述了迄今为止中医药、针灸治疗男性不育的相关进展及针灸的治疗机理;总结了现代医学对男性不育少弱精症的病因及病理机制的认识,以及药物治疗本病的进展。针灸治疗基于辨证取穴、循经取穴,远端取穴与局部取穴相配合的原则,结合针刺的补泻手法,在临床治疗男性不育症有较好的疗效。本文在前人研究的基础上,结合导师赖新生多年对男性不育症的治疗经验,选取关元、肾俞及三阴交作为治疗用穴,采用电针和艾灸不同的疗法,观察针灸对少弱精大鼠模型生殖功能的影响。
研究方法
46只健康成年SD大鼠,随机抽取10只作为正常对照组,其余36只随机分为三组,分别是造模组、电针组和艾灸组。
造模:将腺嘌呤配成浓度为14mg/ml的混悬液,给SD大鼠灌胃,即20mg/100g体重,每日灌胃1次,共28天。造成大鼠少弱精模型。
治疗:于造模停止的第二天起对电针组大鼠施以电针治疗,取穴关元、肾俞、三阴交,采用疏密波,频率为14次/分钟,强度以大鼠安静耐受为度,留针20分钟,连续治疗45天。同时于造模停止的第二天起给予艾灸组大鼠艾灸治疗,取穴关元和肾俞,每次15分钟,每日治疗一次,连续治疗45天。
检测指标及方法:治疗结束后采用计算机辅助精子质量分析(computer-aided semen analysis,CASA)系统检测大鼠的精子密度、精子活率、前向运动精子百分率;检测大鼠的体重、睾丸及附睾的质量,计算睾丸及附睾的系数;采用放免法检测大鼠血清睾酮(Testosterone,T)、卵
Infertility is a world widely problem influencing people and their families. The number of infertility has amounted to 50000000~80000000, and this kind of disorder has been giving rise to misfortune and grief to people. Additionally, there are 2000000 persons more each year suffering infertility. Moreover, this number increases continuously. Among patients, 40%~50% is due to male factors, 30% of which is caused by disturbance of spermatozoa generation. The situation in China is similar which is not optimistic. Oligospermia refers to the amount of sperms is less than 2 × 10~7/ml;Asthenspermia means number of spermatozoa moving ahead (including grade a and grade b) is less than 50% or the number of sperms grade a less than 25%. Oligospermia and asthenspermia are commonly seen in combination in clinic. The etiology and pathogenesis are very complex which including genetic factors, endocrine, immune factors, spermophlebectasia, infection factor, radiation, chemical materials contact, which affect spermatogenesis and mature. This thesis summarized knowledge of ancient TCM scholars' viewpoints about etiology, pathology and differentiation of syndromes and development of modern treatment of TCM (including acupuncture and moxibustion) and knowledge of etiology, pathology and internal medicine treatment of modern west medicine. It is reported that acupuncture and moxibustion had good clinical effects on male sterility based on principles of locations of points according to differentiation of syndromes, following meridians, combination of
distal position and local position. The points of Guanyuan, Shenshu and Sanyinjiao were adopted in this research, electricity and moxibustion were also applied in treatment to observe the influence on reproductive function of oligospermia and asthenospermia rats animals.MethodsThere were 46 SD rats which were randomized into normal group(10), model group, electro-acupuncture group and moxibustion group( 34 rats in three groups)Establishing models: Confected the suspensoid agent with adenine and sterile water in concentration of 14mg/ml. Gave rats the suspensoid agent with 20mg/100g body weight ? day for 28 days.Treatment: One group was treated by electro-acupunture. Acupuncture the points of Guanyuan, Shenshu and Sanyinjiao for 20minutes with dense-sparse wave in frequency of 14 times/minute. Repeated the treatment every day for 45 days. The other group was treated for 15minutes by moxibustion to the points of Guanyuan and Shenshu. Repeated the treatment every day for 45 days.Detecting indexes: Detected the spermatozoa density, spermatozoa livability and percentage of spermatozoa moving ahead with computer -aided semen analysis(CASA);weight of rat body, weight and quotiety of testicle and epididymis;serum Testosterone (T)> Follicle stimulating homone(FSH)and Luteinizing hormone(LH) with RIA;1C, 2C, 4C 1C/2C, 2C/4C and 1C/4C of testicle tissue by the method of flow cytometry (FCM);integrality of membrane of spermatozo by water test;MDA by colorimetric method with spectrophotometer.All data was processed by software package SPSS 11.5 and analyzed by one-way ANOVA method between four groups comparison and LSD or Games-Howell method between two groups comparison, a =0.05.Results1 The influence on quality of semen by acupuncture and moxibustion1. 1 The result showed: Comparing to normal group, spermatozoa
density in model group, electro-acupuncture group and moxibustion group were decreased (P< 0.01);comparing to model group, spermatozoa density in electro-acupuncture group was significantly higher (P< 0.05);There was not significant difference between moxibustion group and model group.1. 2 Comparing to normal group, spermatozoa livability of model group, electro-acupuncture group and moxibustion group was decreased (P< 0.01);comparing to model group, spermatozoa livability of electro-acupuncture group was significantly higher (P < 0.05);moxibustion group' s was significantly higher than model group(P< 0.05).1.3 Comparing to normal group, percentage of spermatozo moving ahead of model group, electro-acupuncture group and moxibustion group was decreased (P < 0.01);comparing to model group, percentage of spermatozo moving ahead of electro-acupuncture group was significantly higher (P < 0. 05);there was not signif icantl difference between moxibustion group and model group.2 The influence on sexual gland by acupuncture and moxibustion2. 1 Comparing to normal group, the weights and quotieties of testicle of model group, electro-acupuncture group and moxibustion group were decreased (P<0.01);comparing to model group, weight of testicle of electro-acupuncture group was significantly higher (P < 0.05);There was not significant difference between moxibustion group and model group.2.2 Comparing to normal group, the weights and quotieties of epididymis of model group, electro-acupuncture group and moxibustion group were decreased (P < 0.01);the weight and quotietiy of epididymis of electro-acupuncture group model group are significantly higher than that of model group, there were not significant differences between moxibustion group and model group.2.3 Histopathological examination: it showed there was damage to convoluted seminiferous tubules in model group with much less spermatozoa;the damage in electro-acupuncture group was milder than model group;the condition of moxibustion group was similar with model group.
3 The influence on sexual hormone in serum by acupuncture and moxibustionThe result showed: there was not significant difference on serum T between four groups;no significant difference on serum FSH;comparing to normal group, serum LH of model group (P < 0.01), electro-acupuncture group (P < 0. 05) and moxibustion group (P < 0. 01) increased;there was not significant difference between electro-acupuncture group and model group, no significant difference between moxibustion group and model group, but it seemed LH in electro-acupuncture group inclined toward the normal group.4 The influence on seminiferous cell cycle by acupuncture and moxibustion4.1 Comparing to normal group, 1C of model group (P< 0.01), electro-acupuncture group(P < 0.01) and moxibustion group decreased (P < 0. 01);while electro-acupuncture group' s was higher than that of model group(P < 0.05), there was not significant difference between moxibustion group and model group;comparing to normal group, 2C of model group(P< 0.05), electro-acupuncture group(P< 0. 05) and moxibustion group increased (P< 0.05);4.2 Proportion between different seminiferous cells: Comparing to normal group, the value of 4C/2C of model group(P< 0. 05), and moxibustion group increased (P< 0. 05), the value of 1C/4C of model group (P < 0.01), electro-acupuncture group(P < 0.01) and moxibustion group (P < 0.01) decreased significantly, the value of 1C/4C of electro-acupuncture group was higher than that of the value of model group(P < 0.05);Comparing to normal group, 1C/2C of model group(P < 0.01), electro-acupuncture group(P < 0.01) and moxibustion group (P< 0.01) decreased significantly. There were not differences between model group, electro-acupuncture group and moxibustion group.5 The influence on integrality of membrane of spermatozoa tail by acupuncture and moxibustionComparing to normal group, the amount of tail-swollen of spermatozoa of model group(P< 0.01), electro-acupuncture group(P< 0.01) and moxibustion group(P < 0.01) decreased;
electro-acupuncture group' s was higher than that of model group(P < 0.05), there was no difference between model group and moxibustion group.6 The influence on MDA of semen by acupuncture and moxibustionThere was no significant difference on value of MDA of semen between four groups(P > 0.05).ConclusionElectro-acupuncture could promote the reproductive function to certain degree, which was showed in different ways: effectively increasing spermatozo density, spermatozoa livability and percentage of spermatozoa moving ahead;promoting the growth of rats and its' testicle and epididymis;maybe regulating the level of LH;affecting seminiferous cells meiotic division process;protecting the membrane of spermatozoa. Generally, the therapy of moxibustion manifested no obvious effect on seminiferous function of oligospermia and asthenospermia rats except promoting the sperm livability.
研究方法
46只健康成年SD大鼠,随机抽取10只作为正常对照组,其余36只随机分为三组,分别是造模组、电针组和艾灸组。
造模:将腺嘌呤配成浓度为14mg/ml的混悬液,给SD大鼠灌胃,即20mg/100g体重,每日灌胃1次,共28天。造成大鼠少弱精模型。
治疗:于造模停止的第二天起对电针组大鼠施以电针治疗,取穴关元、肾俞、三阴交,采用疏密波,频率为14次/分钟,强度以大鼠安静耐受为度,留针20分钟,连续治疗45天。同时于造模停止的第二天起给予艾灸组大鼠艾灸治疗,取穴关元和肾俞,每次15分钟,每日治疗一次,连续治疗45天。
检测指标及方法:治疗结束后采用计算机辅助精子质量分析(computer-aided semen analysis,CASA)系统检测大鼠的精子密度、精子活率、前向运动精子百分率;检测大鼠的体重、睾丸及附睾的质量,计算睾丸及附睾的系数;采用放免法检测大鼠血清睾酮(Testosterone,T)、卵
Infertility is a world widely problem influencing people and their families. The number of infertility has amounted to 50000000~80000000, and this kind of disorder has been giving rise to misfortune and grief to people. Additionally, there are 2000000 persons more each year suffering infertility. Moreover, this number increases continuously. Among patients, 40%~50% is due to male factors, 30% of which is caused by disturbance of spermatozoa generation. The situation in China is similar which is not optimistic. Oligospermia refers to the amount of sperms is less than 2 × 10~7/ml;Asthenspermia means number of spermatozoa moving ahead (including grade a and grade b) is less than 50% or the number of sperms grade a less than 25%. Oligospermia and asthenspermia are commonly seen in combination in clinic. The etiology and pathogenesis are very complex which including genetic factors, endocrine, immune factors, spermophlebectasia, infection factor, radiation, chemical materials contact, which affect spermatogenesis and mature. This thesis summarized knowledge of ancient TCM scholars' viewpoints about etiology, pathology and differentiation of syndromes and development of modern treatment of TCM (including acupuncture and moxibustion) and knowledge of etiology, pathology and internal medicine treatment of modern west medicine. It is reported that acupuncture and moxibustion had good clinical effects on male sterility based on principles of locations of points according to differentiation of syndromes, following meridians, combination of
distal position and local position. The points of Guanyuan, Shenshu and Sanyinjiao were adopted in this research, electricity and moxibustion were also applied in treatment to observe the influence on reproductive function of oligospermia and asthenospermia rats animals.MethodsThere were 46 SD rats which were randomized into normal group(10), model group, electro-acupuncture group and moxibustion group( 34 rats in three groups)Establishing models: Confected the suspensoid agent with adenine and sterile water in concentration of 14mg/ml. Gave rats the suspensoid agent with 20mg/100g body weight ? day for 28 days.Treatment: One group was treated by electro-acupunture. Acupuncture the points of Guanyuan, Shenshu and Sanyinjiao for 20minutes with dense-sparse wave in frequency of 14 times/minute. Repeated the treatment every day for 45 days. The other group was treated for 15minutes by moxibustion to the points of Guanyuan and Shenshu. Repeated the treatment every day for 45 days.Detecting indexes: Detected the spermatozoa density, spermatozoa livability and percentage of spermatozoa moving ahead with computer -aided semen analysis(CASA);weight of rat body, weight and quotiety of testicle and epididymis;serum Testosterone (T)> Follicle stimulating homone(FSH)and Luteinizing hormone(LH) with RIA;1C, 2C, 4C 1C/2C, 2C/4C and 1C/4C of testicle tissue by the method of flow cytometry (FCM);integrality of membrane of spermatozo by water test;MDA by colorimetric method with spectrophotometer.All data was processed by software package SPSS 11.5 and analyzed by one-way ANOVA method between four groups comparison and LSD or Games-Howell method between two groups comparison, a =0.05.Results1 The influence on quality of semen by acupuncture and moxibustion1. 1 The result showed: Comparing to normal group, spermatozoa
density in model group, electro-acupuncture group and moxibustion group were decreased (P< 0.01);comparing to model group, spermatozoa density in electro-acupuncture group was significantly higher (P< 0.05);There was not significant difference between moxibustion group and model group.1. 2 Comparing to normal group, spermatozoa livability of model group, electro-acupuncture group and moxibustion group was decreased (P< 0.01);comparing to model group, spermatozoa livability of electro-acupuncture group was significantly higher (P < 0.05);moxibustion group' s was significantly higher than model group(P< 0.05).1.3 Comparing to normal group, percentage of spermatozo moving ahead of model group, electro-acupuncture group and moxibustion group was decreased (P < 0.01);comparing to model group, percentage of spermatozo moving ahead of electro-acupuncture group was significantly higher (P < 0. 05);there was not signif icantl difference between moxibustion group and model group.2 The influence on sexual gland by acupuncture and moxibustion2. 1 Comparing to normal group, the weights and quotieties of testicle of model group, electro-acupuncture group and moxibustion group were decreased (P<0.01);comparing to model group, weight of testicle of electro-acupuncture group was significantly higher (P < 0.05);There was not significant difference between moxibustion group and model group.2.2 Comparing to normal group, the weights and quotieties of epididymis of model group, electro-acupuncture group and moxibustion group were decreased (P < 0.01);the weight and quotietiy of epididymis of electro-acupuncture group model group are significantly higher than that of model group, there were not significant differences between moxibustion group and model group.2.3 Histopathological examination: it showed there was damage to convoluted seminiferous tubules in model group with much less spermatozoa;the damage in electro-acupuncture group was milder than model group;the condition of moxibustion group was similar with model group.
3 The influence on sexual hormone in serum by acupuncture and moxibustionThe result showed: there was not significant difference on serum T between four groups;no significant difference on serum FSH;comparing to normal group, serum LH of model group (P < 0.01), electro-acupuncture group (P < 0. 05) and moxibustion group (P < 0. 01) increased;there was not significant difference between electro-acupuncture group and model group, no significant difference between moxibustion group and model group, but it seemed LH in electro-acupuncture group inclined toward the normal group.4 The influence on seminiferous cell cycle by acupuncture and moxibustion4.1 Comparing to normal group, 1C of model group (P< 0.01), electro-acupuncture group(P < 0.01) and moxibustion group decreased (P < 0. 01);while electro-acupuncture group' s was higher than that of model group(P < 0.05), there was not significant difference between moxibustion group and model group;comparing to normal group, 2C of model group(P< 0.05), electro-acupuncture group(P< 0. 05) and moxibustion group increased (P< 0.05);4.2 Proportion between different seminiferous cells: Comparing to normal group, the value of 4C/2C of model group(P< 0. 05), and moxibustion group increased (P< 0. 05), the value of 1C/4C of model group (P < 0.01), electro-acupuncture group(P < 0.01) and moxibustion group (P < 0.01) decreased significantly, the value of 1C/4C of electro-acupuncture group was higher than that of the value of model group(P < 0.05);Comparing to normal group, 1C/2C of model group(P < 0.01), electro-acupuncture group(P < 0.01) and moxibustion group (P< 0.01) decreased significantly. There were not differences between model group, electro-acupuncture group and moxibustion group.5 The influence on integrality of membrane of spermatozoa tail by acupuncture and moxibustionComparing to normal group, the amount of tail-swollen of spermatozoa of model group(P< 0.01), electro-acupuncture group(P< 0.01) and moxibustion group(P < 0.01) decreased;
electro-acupuncture group' s was higher than that of model group(P < 0.05), there was no difference between model group and moxibustion group.6 The influence on MDA of semen by acupuncture and moxibustionThere was no significant difference on value of MDA of semen between four groups(P > 0.05).ConclusionElectro-acupuncture could promote the reproductive function to certain degree, which was showed in different ways: effectively increasing spermatozo density, spermatozoa livability and percentage of spermatozoa moving ahead;promoting the growth of rats and its' testicle and epididymis;maybe regulating the level of LH;affecting seminiferous cells meiotic division process;protecting the membrane of spermatozoa. Generally, the therapy of moxibustion manifested no obvious effect on seminiferous function of oligospermia and asthenospermia rats except promoting the sperm livability.
引文
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