吸烟和解脲支原体感染影响精子质量的相关研究
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摘要
本文主要探讨吸烟、解脲支原体感染对精子质量的影响，并初步探索其相关机制。对不育患者精子参数进行分析，研究吸烟、解脲支原体感染对精子参数（精子形态、精子活力、精子活率、精子运动参数、精子密度，等）的影响，初步探讨精浆锌、白细胞、精子DNA碎片、精浆中性α-葡糖苷酶在吸烟、解脲支原体感染影响精子参数中的作用，探讨其作用机制。研究结果表明，吸烟降低精浆锌水平，降低抗氧化能力，而降低精子活率、活力，增加精子畸形率；吸烟增加精液白细胞，可能通过氧化应激，降低精子质量；吸烟可损伤精子，导致精子DNA碎片增加，降低精子活力，影响精子形态；吸烟不通过降低精浆中性α-葡糖苷酶影响精子质量。解脲支原体改变精子形态，降低受精能力，还可通过增加精液白细胞降低精子质量。本文为吸烟、解脲支原体感染影响精子质量提供了实验依据，对吸烟、解脲支原体感染影响精子质量的机制进行探讨，为男性不育的病因学提供了理论基础，并为孕前咨询、临床治疗及药物开发提供实验依据。
Background
Male infertility, which accounts for20%of the causes of infertility haunting15%ofthe married couples, is an important direction in the subject of reproduction. Other causesof infertility, including disorders due to female alone or both, and the unknow causes,accounting for40%,25%and15%, respectively. Worse still, prevalence of infertility isascending over recent years due to unfavorable life-style, aggravated environmentalpollution and increase of sextually transmitted diseases. Along with research advancing,changes in sperm parameters, due to many influential factors, has recently been found to bethe key to male infertility. Negative effects have been found at smoking to sperm quality inmany researches, but what changes exist in sperm parameters is controversy. And it isundefined that through which way sperm quality is influenced. It needs to be confirmed thatwhether Ureaplasma Urealyticum infection decreases sperm quality and what is themechanism between them.
Objective
This present study aimed at exploring the effects of smoking and UreaplasmaUrealyticum (UU) infection on sperm parameters, including concentration of sperm cells,motility, movement and morphous of sperm cells. And investigating whether smokingeffects seminal plasma zinc, seminal leukocytes concentration, sperm DNA fragmentationand neutral α-glycosidase of seminal plasma, and their role in effects of smoking on spermparameters. In this study, the relationship between UU infection and seminal leukocytes isalso investigated, further more, we also investigate the role of seminal leukocytes in effectsof UU infection on sperm parameters. In order to pave the way for make clear of themechanism involved in male infertility and to provide experimental evidence fordifferential diagnosis and prophylaxis in clinic work.
Methods
During2007to2011,2492male patients suffering from infertility who treated inaffiliated hospitals of Jilin University were enrolled in this study, and semen parametersand sperm morphology analysis were performed.
Firstly,1525male patients suffering from infertility who not infects UU is assignedinto non-smoking group and smoking group which were further stratified into mild subgroup (smoking frequency [SF]: less than5cigarets per day; smoking ages [SA]: lessthan5years), moderate sub-group (SF:5to20cigarets per day; SA:5to10years), andsevere sub-groups (SF: more than20cigarets per day; SA: more than10years). The spermparameters, including concentration of sperm cells, motility, movement and morphous ofsperm cells, as well as seminal leukocytes concentration were detected for all patientswhether smoke or not. In addition, seminal plasma zinc, sperm DNA fragmentation andneutral α-glycosidase of seminal plasma were detected for some of these patients.According to the level of seminal plasma zinc in smokers and non-smokers,147subjectswere additionally assigned into normal sub-group with seminal plasma zinc level more than2.11mmol/L, and abnormal sub-group with seminal plasma zinc level of2.11mmol/L orlower. While136subjects were assigned into normal sub-group with sperm DNAfragmentation of25%or less, and abnormal sub-group with the rate more than25%inregard to sperm DNA fragmentation in smokers and non-smokers. The seminal plasmaα-glucoside content in smokers and non-smokers was20mU per ejaculation or more,according to this standard,140subjects were assigned into normal sub-group and abnormalsub-group.
In addition, in regard to the results of UU infection detected,967subjetcs who arenon-smokers were assigned into UU-positive group and UU-negtive group. Then seminalleukocytes concentration were detected for these subjects. In regard to the results detected,the UU-negative group were divided into normal sub-group with leukocytes less than1×106/ml, and abnormal sub-group with leukocytes concentration of1×106/ml or more.And sperm parameters such as concentration of sperm cells, motility, movement of spermcells were also detected for these subjects. According to whether the concentration ofseminal leukocytes was normal or not, the UU-negative group and UU-positive group wasdivided into normal sub-group and abnormal sub-group.
The statistical methods t-test, nonparametric t-test, χ2-test, correlation analysis andanalysis of variance were applied to study.
Results
Smokers had a significant lower proportions of fast (P<0.01) and slow (P<0.05)progressive, sperm motility (P<0.01) and sperm viability (P<0.05) as compared with nosmoking group. Statistical difference was found in Sperm movement parameters, includingVAP、 LIN、 STR(P<0.05)、 WOB(P<0.01) which were lower in smokers than innon-smokers. Sperm normal morphology percentage was lower in smokers than in non-smokers (P<0.01), Morphologically, percentage of abnormal sperm, including spermwith defects especially those with the head defects (P<0.05) were more. In comparison withnon-smokers, a decrease was seen in slow progressive sperms (P<0.05) of mild smokers, inpercentage of sperm motility and fast progressive sperms (P<0.05)of moderate smokers, inpercentage of sperm motility, viability and fast progressive sperms of severe smokers(P<0.05), whereas an increase was seen in percentage of non-progressive sperms (P<0.05).When compared with non-smokers, the sperm motion parameters were no change and inmoderate with VCL、VAP、LIN、WOB、STR (P <0.05) lower and severe smokers withrespect to VCL、LIN、WOB、STR (P<0.05) and VAP、VSL (P<0.01) lower. Compared withnonsmokers, the sperm morphology was no significant abnormal in mild smokers, butnormal sperms were less in moderate (P<0.05) and severe (P<0.01) smokers, especiallyhead defect (P<0.05, P<0.01). The concentration of seminal plasma zinc in smokers wasless than non-smokers (P<0.05). An inverse relation existed between smoking years andquantity and seminal plasma zinc concentration(r=-0.335, P<0.01；r=-0.438, P<0.01),Sperm parameters (concentration, motility and morphology) were found to be significantlylower among smokers than non-smokers (P<0.05). A decrease was found to be significantlygreater in smokers with abnormal seminal plasma zinc levels when compared to smokerswith normal seminal plasma zinc levels (P<0.05) or with either group of non-smokers(P<0.01).The leukocyte levels in semen of smoking and non-smoking groups were less than1×106/ml, and its level of smoking group was higher than non-smoking group. The rise ofleukocyte level existed in the moderate (P<0.05) and severe (P<0.01) smoking group,especially in the later one. The sperm DNA fragmentation of smoking group was higherthan non-smoking group (P<0.05). A positive relation existed between smoking years andquantity and sperm DNA fragmentation (r=0.436, P<0.01；r=0.263, P<0.05) Spermparameters (motility and morphology) were found to be significantly lower amongnon-smokers with abnormal high sperm DNA fragmentation than normal ones (P<0.05)and smokers with abnormal high sperm DNA fragmentation than non-smokers withabnormal sperm DNA fragmentation and smokers with normal ones(P<0.05；P<0.01) andsmokers with normal sperm DNA fragmentation (P<0.05).The neutral α-glycosidase ofseminal plasma was no significant different between smoking group and non-smokinggroup. The sperm parameters were similar among the normal and abnormal neutralα-glycosidase of seminal plasma in smoker and non-smoker.
UU-positive group had a significantly higher proportion of abnormal sperm ascompared with UU-negative infertile group (P<0.05). The percentage of sperm cells withhead and tail defects was significantly higher in UU-positive infertile group than that inUU-negative infertile groups (P<0.01and P<0.05, respectively). An abnormally highleucocytes count (>1×106/ml of semen) was observed in a higher percentage ofUU-positive infertile patients than UU-negative infertile patients (P<0.001).The patientswith abnormally high seminal leukocytes were found to have a significantly decreasedpercentage of fast and slow progressive sperms (P<0.01and P<0.05, respectively), spermmotility and viability (P<0.01), VCL, VSL, VAP, LIN, WOB, STR (P<0.001), andincreased percentage of non-progressive sperms (P<0.01). Infertile patients withUU-positive have an abnormally increase in high seminal leukocytes than them withUU-negtive (χ2=43.67, P<0.001). Infertile patients with abnormally high seminalleukocytes were found to have a significantly increased percentage of morphologicallyabnormal sperm, as compared with infertile patients with normal leucocytes levels (P<0.01,P<0.05, respectively). The percentage of head and tail defects in which had significantlyincrease (P<0.05). The percentage of abnormal sperm was significantly higher inUU-positive patients with abnormally high seminal leucocytes as compared withUU-negative men with abnormally high seminal leucocytes (P<0.05). The percentage ofhead defects in which had significantly increase (P<0.05).
Conclusion
Along with smoking, lots of harmful substances such as nicotine increase in spermmalformation and lead to decrease sperm fertilization ability. With prolongation of time andincrease in numbers of cigarettes, adverse effects of smoking accumulate in adose-dependant manner. Meanwhile, a decrease in level of seminal zinc which plays apivotal role in oxidative damage, and further influence the quality of sperm. What is more,excessive product was produced by leukocytes the number of which in seminal fluid wasraised during smoking, which attributes to decrease of movement of sperm, and worse still.The sperm DNA damage was worse and cause to sperm parameters decrease by smoking.Seminal plasma neutral α-glycosidase was not changed by smoking to decrease spermquality. Briefly speaking, cigarette smoking may do harm to sperm in three ways. Firstly,decrease in seminal zinc level leads to attenuation of anti-oxidative ability and influence inquality, activity and morphology of sperm, and at last, the male fertility. Secondly, seminal leucocytes increase abnormally, which leads to oxidation stress causing abnormalities insperm parameters. Finally,.harmful substances exert adverse effects on sperms, leading tosperm DNA damage, sperm paramorphia increase and sperm fertilization ability decrease.
UU increase in sperm malformation, and therefore, the fertilizability of sperm isweakened. UU infection leads to lipid peroxidation injuries of chondriosome, decrease insynthesis of ATP and sperm activity, and attentuation of forward movement ability.Moreover, impaired antioxidant system initiate oxidative stress may reduce quality ofsperm though diminishing the percentage of fast and slow progressive sperm, motility andsurvial rate and sperm motor ability. As for the mechanisms affecting the quality of sperm,one is UU exert adverse effects on sperms, leading to sperm paramorphia increase andsperm fertilization ability decrease, the other one referred to the abnormities of leukocytes,which influence either the sperm activity to cause male infertility.
Background
Male infertility, which accounts for20%of the causes of infertility haunting15%ofthe married couples, is an important direction in the subject of reproduction. Other causesof infertility, including disorders due to female alone or both, and the unknow causes,accounting for40%,25%and15%, respectively. Worse still, prevalence of infertility isascending over recent years due to unfavorable life-style, aggravated environmentalpollution and increase of sextually transmitted diseases. Along with research advancing,changes in sperm parameters, due to many influential factors, has recently been found to bethe key to male infertility. Negative effects have been found at smoking to sperm quality inmany researches, but what changes exist in sperm parameters is controversy. And it isundefined that through which way sperm quality is influenced. It needs to be confirmed thatwhether Ureaplasma Urealyticum infection decreases sperm quality and what is themechanism between them.
Objective
This present study aimed at exploring the effects of smoking and UreaplasmaUrealyticum (UU) infection on sperm parameters, including concentration of sperm cells,motility, movement and morphous of sperm cells. And investigating whether smokingeffects seminal plasma zinc, seminal leukocytes concentration, sperm DNA fragmentationand neutral α-glycosidase of seminal plasma, and their role in effects of smoking on spermparameters. In this study, the relationship between UU infection and seminal leukocytes isalso investigated, further more, we also investigate the role of seminal leukocytes in effectsof UU infection on sperm parameters. In order to pave the way for make clear of themechanism involved in male infertility and to provide experimental evidence fordifferential diagnosis and prophylaxis in clinic work.
Methods
During2007to2011,2492male patients suffering from infertility who treated inaffiliated hospitals of Jilin University were enrolled in this study, and semen parametersand sperm morphology analysis were performed.
Firstly,1525male patients suffering from infertility who not infects UU is assignedinto non-smoking group and smoking group which were further stratified into mild subgroup (smoking frequency [SF]: less than5cigarets per day; smoking ages [SA]: lessthan5years), moderate sub-group (SF:5to20cigarets per day; SA:5to10years), andsevere sub-groups (SF: more than20cigarets per day; SA: more than10years). The spermparameters, including concentration of sperm cells, motility, movement and morphous ofsperm cells, as well as seminal leukocytes concentration were detected for all patientswhether smoke or not. In addition, seminal plasma zinc, sperm DNA fragmentation andneutral α-glycosidase of seminal plasma were detected for some of these patients.According to the level of seminal plasma zinc in smokers and non-smokers,147subjectswere additionally assigned into normal sub-group with seminal plasma zinc level more than2.11mmol/L, and abnormal sub-group with seminal plasma zinc level of2.11mmol/L orlower. While136subjects were assigned into normal sub-group with sperm DNAfragmentation of25%or less, and abnormal sub-group with the rate more than25%inregard to sperm DNA fragmentation in smokers and non-smokers. The seminal plasmaα-glucoside content in smokers and non-smokers was20mU per ejaculation or more,according to this standard,140subjects were assigned into normal sub-group and abnormalsub-group.
In addition, in regard to the results of UU infection detected,967subjetcs who arenon-smokers were assigned into UU-positive group and UU-negtive group. Then seminalleukocytes concentration were detected for these subjects. In regard to the results detected,the UU-negative group were divided into normal sub-group with leukocytes less than1×106/ml, and abnormal sub-group with leukocytes concentration of1×106/ml or more.And sperm parameters such as concentration of sperm cells, motility, movement of spermcells were also detected for these subjects. According to whether the concentration ofseminal leukocytes was normal or not, the UU-negative group and UU-positive group wasdivided into normal sub-group and abnormal sub-group.
The statistical methods t-test, nonparametric t-test, χ2-test, correlation analysis andanalysis of variance were applied to study.
Results
Smokers had a significant lower proportions of fast (P<0.01) and slow (P<0.05)progressive, sperm motility (P<0.01) and sperm viability (P<0.05) as compared with nosmoking group. Statistical difference was found in Sperm movement parameters, includingVAP、 LIN、 STR(P<0.05)、 WOB(P<0.01) which were lower in smokers than innon-smokers. Sperm normal morphology percentage was lower in smokers than in non-smokers (P<0.01), Morphologically, percentage of abnormal sperm, including spermwith defects especially those with the head defects (P<0.05) were more. In comparison withnon-smokers, a decrease was seen in slow progressive sperms (P<0.05) of mild smokers, inpercentage of sperm motility and fast progressive sperms (P<0.05)of moderate smokers, inpercentage of sperm motility, viability and fast progressive sperms of severe smokers(P<0.05), whereas an increase was seen in percentage of non-progressive sperms (P<0.05).When compared with non-smokers, the sperm motion parameters were no change and inmoderate with VCL、VAP、LIN、WOB、STR (P <0.05) lower and severe smokers withrespect to VCL、LIN、WOB、STR (P<0.05) and VAP、VSL (P<0.01) lower. Compared withnonsmokers, the sperm morphology was no significant abnormal in mild smokers, butnormal sperms were less in moderate (P<0.05) and severe (P<0.01) smokers, especiallyhead defect (P<0.05, P<0.01). The concentration of seminal plasma zinc in smokers wasless than non-smokers (P<0.05). An inverse relation existed between smoking years andquantity and seminal plasma zinc concentration(r=-0.335, P<0.01；r=-0.438, P<0.01),Sperm parameters (concentration, motility and morphology) were found to be significantlylower among smokers than non-smokers (P<0.05). A decrease was found to be significantlygreater in smokers with abnormal seminal plasma zinc levels when compared to smokerswith normal seminal plasma zinc levels (P<0.05) or with either group of non-smokers(P<0.01).The leukocyte levels in semen of smoking and non-smoking groups were less than1×106/ml, and its level of smoking group was higher than non-smoking group. The rise ofleukocyte level existed in the moderate (P<0.05) and severe (P<0.01) smoking group,especially in the later one. The sperm DNA fragmentation of smoking group was higherthan non-smoking group (P<0.05). A positive relation existed between smoking years andquantity and sperm DNA fragmentation (r=0.436, P<0.01；r=0.263, P<0.05) Spermparameters (motility and morphology) were found to be significantly lower amongnon-smokers with abnormal high sperm DNA fragmentation than normal ones (P<0.05)and smokers with abnormal high sperm DNA fragmentation than non-smokers withabnormal sperm DNA fragmentation and smokers with normal ones(P<0.05；P<0.01) andsmokers with normal sperm DNA fragmentation (P<0.05).The neutral α-glycosidase ofseminal plasma was no significant different between smoking group and non-smokinggroup. The sperm parameters were similar among the normal and abnormal neutralα-glycosidase of seminal plasma in smoker and non-smoker.
UU-positive group had a significantly higher proportion of abnormal sperm ascompared with UU-negative infertile group (P<0.05). The percentage of sperm cells withhead and tail defects was significantly higher in UU-positive infertile group than that inUU-negative infertile groups (P<0.01and P<0.05, respectively). An abnormally highleucocytes count (>1×106/ml of semen) was observed in a higher percentage ofUU-positive infertile patients than UU-negative infertile patients (P<0.001).The patientswith abnormally high seminal leukocytes were found to have a significantly decreasedpercentage of fast and slow progressive sperms (P<0.01and P<0.05, respectively), spermmotility and viability (P<0.01), VCL, VSL, VAP, LIN, WOB, STR (P<0.001), andincreased percentage of non-progressive sperms (P<0.01). Infertile patients withUU-positive have an abnormally increase in high seminal leukocytes than them withUU-negtive (χ2=43.67, P<0.001). Infertile patients with abnormally high seminalleukocytes were found to have a significantly increased percentage of morphologicallyabnormal sperm, as compared with infertile patients with normal leucocytes levels (P<0.01,P<0.05, respectively). The percentage of head and tail defects in which had significantlyincrease (P<0.05). The percentage of abnormal sperm was significantly higher inUU-positive patients with abnormally high seminal leucocytes as compared withUU-negative men with abnormally high seminal leucocytes (P<0.05). The percentage ofhead defects in which had significantly increase (P<0.05).
Conclusion
Along with smoking, lots of harmful substances such as nicotine increase in spermmalformation and lead to decrease sperm fertilization ability. With prolongation of time andincrease in numbers of cigarettes, adverse effects of smoking accumulate in adose-dependant manner. Meanwhile, a decrease in level of seminal zinc which plays apivotal role in oxidative damage, and further influence the quality of sperm. What is more,excessive product was produced by leukocytes the number of which in seminal fluid wasraised during smoking, which attributes to decrease of movement of sperm, and worse still.The sperm DNA damage was worse and cause to sperm parameters decrease by smoking.Seminal plasma neutral α-glycosidase was not changed by smoking to decrease spermquality. Briefly speaking, cigarette smoking may do harm to sperm in three ways. Firstly,decrease in seminal zinc level leads to attenuation of anti-oxidative ability and influence inquality, activity and morphology of sperm, and at last, the male fertility. Secondly, seminal leucocytes increase abnormally, which leads to oxidation stress causing abnormalities insperm parameters. Finally,.harmful substances exert adverse effects on sperms, leading tosperm DNA damage, sperm paramorphia increase and sperm fertilization ability decrease.
UU increase in sperm malformation, and therefore, the fertilizability of sperm isweakened. UU infection leads to lipid peroxidation injuries of chondriosome, decrease insynthesis of ATP and sperm activity, and attentuation of forward movement ability.Moreover, impaired antioxidant system initiate oxidative stress may reduce quality ofsperm though diminishing the percentage of fast and slow progressive sperm, motility andsurvial rate and sperm motor ability. As for the mechanisms affecting the quality of sperm,one is UU exert adverse effects on sperms, leading to sperm paramorphia increase andsperm fertilization ability decrease, the other one referred to the abnormities of leukocytes,which influence either the sperm activity to cause male infertility.
引文
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