【作者】 陈昱；

【导师】 严孙杰；

【作者基本信息】 福建医科大学 ， 内科学， 2010， 硕士

【摘要】 【第一部分】低强度微波辐射对雄性小鼠生殖功能的影响【目的】探讨低强度微波辐射对雄性小鼠生殖功能的影响。【方法】4周龄雄性昆明种小鼠45只,随机分为4组,用相当于移动电话辐射频率的模拟辐射源(900MHz)对小鼠进行全身辐射,辐照平均功率密度分别为0μW/cm2(伪照射组)、50μW/cm2、150μW/cm2、250μW/cm2组,每天12小时,连续60d后取双侧睾丸和附睾称重,常规分析附睾的精子计数、活动率和畸形率,并观察睾丸组织结构,及测定血清睾酮、黄体生成素水平。【结果】(1)各组小鼠脏器系数:各照射组小鼠睾丸重、睾丸指数均有不同程度的降低,其中50μW/cm2组为(84.96±20.58)mg、(0.174±0.039)% ;150μW/cm2组为(6.29±21.6)mg、(0.161±0.046)%;与伪照射组的(138.45±30.34)mg、(0.287±0.056)%相比具有统计学显著性差别(P<0.01)。(2)精子的常规分析结果:50μW/cm2组、150μW/cm2组、250μW/cm2组的精子平均活动率分别为59.625%、44.917%、53.545%,与伪照射组(78.750%)比较,差异均具有显著性(P<0.01);50μW/cm2组、150μW/cm2组、250μW/cm2组的精子平均畸形率分别为4.52%、7.192%、4.673%,与伪照射组(2.840%)比较,差异均具有显著性(P <0.01);精子相对计数与伪照射组[(6.51±0.74)×106/ml]比较, 50μW/cm2组为(5.35±0.75)×106/ml(P <0.01);150μW/cm2组为(5.43±1.00)×106/ml(P <0.01),差别有显著性。250μW/cm2组的精子相对计数较伪照射组稍低,但差别无显著意义。(3)睾丸组织结构观察结果:光镜下可见各微波辐照组小鼠睾丸组织曲细精管上皮结构稍疏松,曲细精管内成熟精子稀少(50μW/cm2、250μW/cm2组)和缺失(150μW/cm2组),呈生精减少或停滞;生精小管间的间质细胞减少。以上变化以150μW/cm2组最明显,250μW/cm2组次之, 50μW/cm2组也有明显的改变。(4)各组血清激素水平比较:50μW/cm2、150μW/cm2照射组血清睾酮水平[(2.546±0.535)ng/mL、(2.093±0.732)ng/mL]较伪照射组[(3.466±0.808) ng/mL]有明显的降低,250μW/cm2照射组也呈下降趋势,但与对照组比未达统计学显著水平。各组血清LH水平无差别,但150μW/cm2照射组数值较低。【结论】在本实验条件下,微波辐射可引起实验小鼠精子计数减少、精子活动率下降及精子畸形增多,并可引起血清睾酮水平降低。提示低强度的微波辐射可能对雄性生殖功能产生不良影响。【第二部分】低强度微波辐射对雄性小鼠生殖内分泌功能的影响【目的】探讨低强度微波辐射对雄性小鼠生殖内分泌功能的影响。【方法】48只4周龄雄性昆明种小鼠,随机分为六组,伪照射组、去睾丸对照组、去睾丸50μW/cm2照射组、去睾丸150μW/cm2照射组、去睾丸250μW/cm2照射组和单纯辐照对照组(150μW/cm2)。照射组小鼠处理同第一部分。试验结束后,断头处死小鼠,立即取血与脑组织进行如下分析:各小鼠均测定血清睾酮(T)、黄体生成素水平(LH),测定下丘脑组织琥珀酸脱氢酶(SDH)、ATP酶(ATPase)活力;各组随机抽取3只动物的下丘脑组织进行电镜超微结构检查;伪照射组、单纯照射对照组取双侧睾丸称重,并计算脏器指数。【结果】(1)各组血清激素水平比较:辐照对照组小鼠血清睾酮水平[(3.026±0.367 )ng/mL]较伪照射组(4.256±0.501ng/mL)有明显的降低,差别具有显著意义。各去睾丸组小鼠血清睾酮均处于极低水平。辐照对照组小鼠血清LH水平[(5.480±2.276)mIU/ml]比伪照射组[(6.493±1.872)mIU/ml)有所降低,但差别无显著意义。去睾丸对照组小鼠血清LH水平[(8.614±2.843)mIU/ml]比伪照射组[(6.493±1.872)mIU/ml]有所升高,虽差别未达统计学显著水平,但也显示了去除睾丸带来的影响。去睾丸照射各组血清LH水平分别是:50μW/cm2组为(7.339±2.663)mIU/ml、150μW/cm2组为(6.276±3.020)mIU/ml、250μW/cm2组为(7.663±3.464)mIU/ml。即在微波辐照条件下,各去睾丸组血清LH水平均未升高至去睾丸对照组水平(8.614±2.843)mIU/ml,其中150μW/cm2组只处于伪照射组水平,说明照射各组下丘脑垂体分泌功能受到一定抑制。(2)下丘脑组织琥珀酸脱氢酶、ATP酶活力测定结果:脑SDH活性水平比较,伪照射组小鼠下丘脑组织SDH活性水平[(12.341±3.343)U/mgprot]与去睾丸对照组小鼠[(12.346±2.023)U/mgprot]无差别。而各照射组下丘脑组织SDH活性水平均有明显的降低,辐射对照组为(8.534±1.455 )U/mgprot、50μW/cm2组为(9.074±1.719)U/mgprot、150μW/cm2组为(7.453±1.240)U/mgprot、250μW/cm2组为(8.120±1.663)U/mgprot,与伪照射组和去睾丸对照组相比差别均有统计学意义。脑ATPase活性水平比较,伪照射组小鼠下丘脑组织ATPase活性水平[(30.704±6.057)U/mgprot]与去睾丸对照组小鼠[(29.898±5.378)U/mgprot]无差别。而各照射组下丘脑组织ATPase活性水平均有明显的降低,辐射对照组为(18.651±3.486)U/mgprot、50μW/cm2组为(24.240±4.365)U/mgprot、150μW/cm2组为(16.425±4.153)U/mgprot、250μW/cm2组为(17.911±3.529)U/mgprot,与伪照射组和去睾丸对照组相比均有统计学意义。照射组中,50μW/cm2组、150μW/cm2组血清LH水平与其小鼠下丘脑组织ATP酶活力呈正相关,相关系数分别为0.795、0.717,提示下丘脑垂体细胞分泌功能的改变与ATP酶活力改变有关。(3)电镜下各组下丘脑线粒体形态学观察结果:电镜下各组下丘脑线粒体形态学观察结果:某些照射组下丘脑细胞线粒体密度减少,线粒体出现不同程度的肿胀、嵴变形和消失等的变化,其中150μW/cm2组最为明显。表明该条件照射下,下丘脑线粒体受到损伤。【结论】在本实验条件下,微波辐射造成了小鼠下丘脑组织细胞中与线粒体功能密切相关两种酶活力的下降,并造成一定程度的线粒体损伤,这可能带来细胞供能障碍的后果,影响下丘脑垂体系统的分泌功能,进而影响生殖功能。这些结果为研究长期暴露在低水平的移动电话微波辐射下对男性生殖功能的影响提供了新的线索。更多还原

【Abstract】 Part I Effects of low intensity microwave radiation on reproductive function in male mice【Objective】To investigate the effects of low intensity microwave radiation on reproductive function in male mice.【Methods】45 male mice of 4 weeks age were randomized into 4 groups and exposed to electromagnetic fields with frequency of 900MHz and power density of 0μW/cm2(sham-exposed controls)、50μW/cm2、150μW/cm2、250μW/cm2 respectively 12 hours per day for 60 days using experimental exposure device.On the day of last treatment, immediately after the termination of exposure sham-exposure and exposure mice were sacrificed. The testes and epididymis of every mouse were viscerated and weighted,and the sperm count,sperm motility and morphology of sperm in epididymis were analysed, and Serum testosterone(T) and luteinizing hormone(LH) were measured.【Results】(1) The organ indexThe testes weight and testes index of radiation groups decreased in some degree:The testes weight and testes index of 50μW/cm2 groupe were (84.96±20.58)mg、(0.174±0.039)% respectively,and those of 150μW/cm2 groupe were (76.29±21.66)mg、(0.161±0.046)% respectively.They all lower than those[(138.45±30.34mg、(0.287±0.056)%] of sham-exposed controls statistically significantly . The epididymis weight of each group showed no notable change.(2) Effects of microwave radiation on sperm counting,sperm motility and abnormal sperm rate of male miceThe average sperm motility rates of 50μW/cm2 group、150μW/cm2 group、250μW/cm2 group were 59.625%、44.917%、53.545% respectively,and they all lower than tha(t78.750%)of sham-exposed controls statistically significantly (P<0.01 for all).The average abnormal sperm rates of 50μW/cm2 group、150μW/cm2 group、250μW/cm2 group were 4.52%、7.192%、4.673% respectively,and they all lower than tha(t2.840%)of sham-exposed controls statistically significantly (P<0.01 for all).The average sperm counting of 50μW/cm2 group、150μW/cm2 group were (5.35±0.75)×106/ml and (5.43±1.00)×106/ml respectively,and they were both lower than that[(6.51±0.74)×106/ml] of sham-exposed controls statistically significantly (P<0.05 and P<0.01 respectively). In 250μW/cm2 group, the average sperm counting was (5.95±0.94)×106/ml and was not different from that of sham-exposed group.(3)Histology of testes:In testis pathological section of exposed groups mice,there existed various degrees of confused cell layers of seminiferous tubule and broadened intercellular space.The mount of sperms in seminiferous tubule decreased or even vanished showing a weaken capacity of spermatogenesis. The density of Leydig cell showed some degrees of decrease. The mice in 150μW/cm2 group showed the most significant change, followed by 250μW/cm2 group and 50μW/cm2 group.(4) The level of hormone in serum:The average serum T level of 50μW/cm2 group、150μW/cm2 group mice were (2.546±0.535) ng/ml and ( 2.093±0.732) ng/ml espectively,and they were both lower than that [(3.466±0.808)ng/ml]of sham-exposed controls statistically significantly (P<0.05 and P<0.01 respectively). The average serum T level of 150μW/cm2 group mice was (2.868±0.749) ng/ml, but the difference of it with that of sham-exposed controls was of no statistically significant though the number of the former was somehow lower. Though the serum T level showed difference, the average serum LH level had no distinction among groups.【Conclusions】Under this experimental condition,900MHz microwave radiation at power density range from 50mW/cm2 to 250mW/cm2 may cause testes injury, sperm counting decrease,sperm motility rates decline,and abnormal sperm rates elevating and change serum T level of male mice.This result hints that low intensity of.civilian use microwave band may affect male reproductive function negatively.Part II Effects of low intensity microwave radiation on reproductive endocrine function in male mice【Objective】To investigate the effects of low intensity microwave radiation on reproductive endocrine function in male mice.【Methods】48 male mice of 4 weeks age were randomized into 6 groups. They are sham-exposed control group,caponizing control group, pure radiating control group with microwave radiating of 150μW/cm2, caponizing group with microwave radiating of 50μW/cm2, caponizing group with microwave radiating of 150μW/cm2, and caponizing group with microwave radiating of 250μW/cm2. The radiating groups were whole-body exposure to 900MHz GSM-like microwave 12 hours per day for 60 days using experimental exposure device.On the day of termination, the mice were sacrificed with decapitate. The testes of the mice in sham-exposed controls and pure radiating controls were viscerated and weighted.The serum testosterone(T) level and luteinizing(LH) level were detected for every mouce and compared by group. The SDH and ATPase activities of hypothalamus tissue were measured for every mouce and compared by group. The ultrastructure especially the mitochondria ultrastructure of hypothalamus tissue was investigated electron microscopically.【Results】(1)The organ index:The testes weight and testes index of pure radiating controls were (0.123±0.014)g、(0.263±0.032)% respectively.They both lower than those[(0.147±(0.016)g、(0.303±0.042)%] of sham-exposed controls statistically significantly . But the epididymis weight of pure radiating controls showed no notable change.(2) The level of hormone in serum:The serum testosterone(T) concentrations of all caponizing groups were in very low level. The serum testosterone(T) concentrations of pure radiating controls was (3.026±0.367)ng/mL,and that of sham-exposed controls was (4.256±0.501)ng/mL.They had differance with statistically significance.The serum luteinizing(LH) concentrations of pure radiating controls was (5.480±2.276)mIU/ml,while The serum LH concentrations of sham-exposed controls was (6.493±1.872)mIU/ml.But the difference was not statistically significant.The serum LH concentrations [(8.614±2.843)mIU/ml] of caponizing controls had some elevate compared with that [(6.493±1.872)mIU/ml] of sham-exposed controls.though the difference was not statistically significant ,the phenomeno showed the influence of caponizing on LH level.About the serum LH of microwave radiating caponizing groups, it was (7.339±2.663)mIU/ml in 50μW/cm2group、(6.276±3.020)mIU/ml in 150μW/cm2 group、and (7.663±3.464)mIU/ml in 250μW/cm2 group.This result showed that under the radiation of microwave, the serum LH level of all caponizing groups’mice did not reach the level [(8.614±2.843)mIU/ml] of caponizing controls,and that of 150μW/cm2 group was only as high as that of sham-exposed controls.That indicated that microwave radiation may depress the secretory function of pituitary.(3) The SDH and ATPase activities of hypothalamus tissue:The SDH activities of hypothalamus tissue of sham-exposed controls and caponizing controls were (12.341±3.343)U/mgprot and (12.346±2.023)U/mgprot respectively, and showed no statistically significance.But all radiating exposure groups showed low activity level and had statistically significance compare with that of sham-exposed controls.They were (8.534±1.455)U/mgprot in pure radiating controls,(9.074±1.719)U/mgprot in 50μW/cm2 group,(7.453±1.240)U/mgprot in 150μW/cm2 group,and (8.120±1.663)U/mgprot in 250μW/cm2 group.The ATPase activities of hypothalamus tissue of sham-exposed controls and caponizing controls were (30.704±6.057)U/mgprot and (29.898±5.378)U/mgprot respectively, and their diffrence had no statistically significance. But all radiating exposuregroups showed low activity level and had statistically significance compare with that of sham-exposed controls.They were (18.651±3.486)U/mgprot in pure radiating controls, (24.240±4.365)U/mgprot in 50μW/cm2 group, (16.425±4.153)U/mgprot in 150μW/cm2 group, and (17.911±3.529)U/mgprot in 250μW/cm2 group respectively.The correlations of serum LH level with hypothalamus ATPase activity were 0.795 in 50μW/cm2 group and 0.717 in 150μW/cm2 group respectively with statistically significance, and this phenomenon indicates that there may be some association between serum LH level and hypothalamus function.(4)mitochondria ultrastructure of hypothalamus tissue:In radiation groups, the mitochondria ultrastructure of hypothalamus tissue displayed changes of various degrees,including mitochondria density decling, part mitochondria swelling,decrease or even disappearance of mitochondrial cristae ,and cavitation formation. The most obvious change happened in 150μW/cm2 group.【Conclusions】Under this experimental condition, 900MHz microwave radiation may impair cell mitochondria and energe metabolism of hypothalamus tissue cells and thus influence their physiologic function.These results contribute to the assessment of potential male fertility risks after continuous chronic exposure to RF radiation at low level.更多还原