| Reproduction is the world biological inevitable process of the new and old replacement.For humans,men and women are the main body in the process of reproductive.It will lead to non-reproduction when alternative has problem.At present,the male sterility has been widely concerned.According to the survey,couples who can't pregnant reach up to at least 50% of the abnormal factors from the male reproductive system.The cause of male infertility is more complicated.Accompany the life habits,diet structures and the change of the surrounding environments, influencing factors are also different.In recent years, with the microwave technology widely application in people's life and work,harm to health produced by the microwave radiation has been a growing number of studies confirming.The reproductive system is the sensitive target organ of microwave radiation.A certain frequency of continuous microwave radiation can lead to male infertility.In recent years,although relevant theories of the microwave radiation influencing on the male reproductive system are in perfecting,its mechanism is still unclear.Traditional Chinese medicine Morida Officinalis How is dry root of rubiaceae plant medicinal indianmulberry with functions of kidney Yang, strong bones and muscles.Our previous studies have found that water extract and alcohol extract of Morida Officinalis How have a certain effects on dyszoospermia caused by microwave radiation.In recent years, domestic and foreign scholars have do a certain research in effective components and effects of Morida Officinalis How.As one of the main ingredients,Morida Officinalis How plays an important role in such aspects of antioxidation,enhancing immunity and improving the cognitive behaviors.But effects and mechanism of Morida Officinalis How polysaccharide on reproductive system dysfunction caused by microwave radiation are rare reported.In accordance with the above questions,this topic focuses on protection of Morida Officinalis How polysaccharide on male rat dyszoospermia caused by 1800 MHZ microwave radiation and influences on orexin neurons in the hypothalamus to investigate the mechanism of action of Morida Officinalis How polysaccharide.Three parts of the study were included.Part One Building dyszoospermia models of SD male rats caused by 1800 MHZ microwave radiationObjective:To build dyszoospermia models of SD male rats caused by 1800 MHZ microwave radiationMethod: Male rats were divided into three groups according to completely random grouping method,namely Control(Control) group 10,microwave radiation(MWR) 50 and sham radiation group(Sham) 10.MWR group of male rats from 8:00 am to 8:00 pm daily were exposured to 1800 MHZ microwave radiation in the box with power density of 200 uw/cm2 and 4 weeks.Sham group of male rats from 8:00 am to 8:00 pm daily were under microwave radiation in the box which does not switch on radiation.Control groups of rats in rat cage with no radiation source.Three groups are guaranteed position,feeding and drinking water freely.Mental states,body weights,body hair and activities of three groups of males rats were observated.Indicators of sexual behaviors(numbers of climbing back and ejaculation, latency periods of ejaculation) and sperm parameters(sperm counts and sperm deformity rates) of three groups of male rats were detected.The organizational structures of three groups of male rats' testis and epididymis were observed by HE dyeing and transmission electron microscope.Result: After 2 w and 4 w of radiation,three groups of rats weights had no statistical significance(P> 0.05).MWR group of male rats' skin flabby,body hair withered and yellow,thick, dry,gloss differential with a third part especially obvious after 4 w of radiation.Male rats show manic,trying to flee,irritability,and shoulder arch back behavior.Compared with the control group and Sham group,the incubation periods of climbing back,ejaculatory latency periods in MWR group of male rats were significantly prolonged,while the numbers of ejaculation and climbing back were decreased significantly,and sperm counts,sperm deformity rates were increased significantly with statistical significances(P<0.01).Compared with the control group and Sham group,seminiferous tubule structures of testicular were observed more loosen and wider spacing.The epithelial basement membranes were thinner.Sperm cells were visibly loosen,falling off with almost no leydig cells and sperms few inside lumen in MWR group.Compared with the control group and Sham group,tubule structures of head of epididymis were observed more irregular,smaller diameters and thinner.Sperm cells were visibly loosen,falling off and sperms few inside lumen in MWR group.Conclusion:Sperm production functions and behavior abilities were reduced and testicular damage was caused in male rats when adopting the power density of 200 uw/cm2 1800 MHZ microwave radiation.Dyszoospermia models of SD male rats were estiblished successfully.Part Two Protection of Morida Officinalis How polysaccharide on dyszoospermia models of SD male rats caused by 1800 MHZ microwave radiationObjective:To explore effect of Morida Officinalis How on dyszoospermia models of SD male rats caused by 1800 MHZ microwave radiation. Method: All rats were divided into four groups by random method,namely the control group(the Control group,10),the radiation group(MWR group,10),Morida Officinalis How oligosaccharide group(10),Morida Officinalis How polysaccharide group(30). The Control group had no radiation and drug delivery.MWR group had microwave radiation and 5 m L dual steaming water by gavage administration.Morida Officinalis How oligosaccharide group were given Morida Officinalis How oligosaccharide the next day after microwave radiation by gavage administration(2 times/day,0.03 g/day).Morida Officinalis How polysaccharide groups were given Morida Officinalis How polysaccharide the next day after microwave radiation by gavage administration(100mg/kg?d-1,200mg/kg?d-1 and 300mg/kg?d-1).Those were all with 2 weeks and 5 m L.Wet weights and the coefficients of testis and epididymis were determined in all groups of male rats.Indicators of sexual behaviors and sperm parameters in all groups were detected.Serum levels of luteinizing hormone(LH),follicle-stimulating hormone(FSH) and testosterone(T) were detected in all groups by enzyme immunosorbent(Elisa) method. The organizational structure of three groups of male rats' testis and epididymis were observed by HE dyeing and transmission electron microscope.Result: Wet weights and coefficients of testis and epididymis of male rats in all groups had no statistical significance(P > 0.05).Compared with the control group,incubation periods of climbing back and ejaculation in the MWR group had significantly increased,while numbers of climbing back and ejaculation in MWR group were obviously decreased.Sperm counts significantly reduced while sperm malformation rates increased significantly.All above were with statistical differences(P<0.01).Compared with the MWR group,incubation periods of climbing back and ejaculation in the Morida Officinalis How polysaccharide groups of 200mg/kg and 300mg/kg were decreased obviously,while numbers of climbing back and ejaculation were obviously increased.Sperm counts increased obviously whlie malformation rates significantly reduced.All above were with statistical significances(P<0.01).Compared with the control group,serum levels of FSH and LH in the MWR group were decreased and increased,while T were obviously decreased(P<0.01).Compared with the MWR group,serum levels of T in the Morida Officinalis How polysaccharide groups of 200mg/kg and 300mg/kg were obviously increased(P<0.05).Compared with the MWR group,testicular sperm production epitheliums connected more closely,cellular levels were increased and neat rowed,seminiferous tubule diameters were normal,mount of sperms in the lumen were more.Tissue structures of testicular had no obvious improvement in Morida Officinalis How oligosaccharide group and Morida Officinalis How polysaccharide group of 100mg/kg.There were more closely connection in basement membrane of epithelium,increased lumen cellular levels,normal lumen diameter,sperms more in head of epididymis.Conclusion: Aspects of sperm parameters,sexual behaviors,sex hormone levels and morphology had an improving effect in Morida Officinalis How polysaccharide on dyszoospermia models of SD male rats caused by microwave radiation and 200mg/kg had the best effect.Part Three Influence of Morida Officinalis How polysaccharide on orexin A neuron located in hypothalamus of dyszoospermia models of SD male rats caused by 1800 MHZ microwave radiationObjective:To discuss the possible mechanism of Morida Officinalis How polysaccharide on dyszoospermia models of SD male rats caused by 1800 MHZ microwave radiation.Method: All rats were divided into three groups by random method,namely the control group(the Control group,10),the radiation group(MWR group,10),Morida Officinalis How polysaccharide group(20).The Control group had no radiation and 5 m L dual steaming water by gavage administration.MWR group had microwave radiation of 12h/day for 4 w and 5 m L dual steaming water by gavage administration.Morida Officinalis How polysaccharide groups were given Morida Officinalis How polysaccharide the next day after microwave radiation by gavage administration(200mg/kg?d-1 and 300mg/kg?d-1).Those were all with 2 weeks and 5 m L.Organization structures of hypothalamus and pituitary of all groups were observed by Nissl dyeing.Expression of orexin in hypothalamus and pituitary were observed by immunofluorescent staining.Level of orexin mRNA in hypothalamus was detected by real-time fluorescent quantitative PCR.Feeding and drinking of all rats were detected in metabolism cages.Result:The positive expression in MWR group was less than control one,while those in Morida Officinalis How polysaccharide group of 200mg/kg and 300mg/kg were increased by Nissl dyeing.The pituitary gland tissue in all groups had no evidently positive results.From orexin A immunofluorescent staining in hypothalamus,orexin A neurons mainly located around the fornix and positive fibers are widely distributed.Compared with control group,Orexin A neuron in hypothalamus and Orexin A positve fiber in arcuate nucleus of MWR group were evidently increased(P<0.01).Compared with MWR group,Orexin A neuron in hypothalamus and Orexin A positve fiber in arcuate nucleus of Morida Officinalis How polysaccharide groups of 200mg/kg and 300mg/kg were evidently decreased(P<0.01).There were no Orexin A neuron and Orexin A positve fiber in pituitary.Compared with control group,Orexin A mRNA in hypothalamus of MWR group were evidently increased(P <0.01).Compared with MWR group,Orexin A mRNA in hypothalamus of Morida Officinalis How polysaccharide groups of 200mg/kg and 300mg/kg were evidently decreased with statistically significant differences(P<0.01).Compared with control group,food intake and water quantity in MWR group were evidently increased with statistically significant differences(P<0.01).Compared with MWR group,food intake and water quantity in Morida Officinalis How polysaccharide groups of 200mg/kg and 300mg/kg were evidently increased with statistically significant differences(P<0.01).Conclusion:Morida Officinalis How polysaccharide had the inhibitory effect on orexin A neurons and fibers located in hypothalamus and arcuate nucleus of dyszoospermia models of SD male rats caused by 1800 MHZ microwave radiation.200mg/kg Morida Officinalis How polysaccharide had the best effects,which maybe was one of protection mechanisms for dyszoospermia caused by the microwave radiation. |
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