纳米细菌对人精子质量的影响

Effect of nanobacteria on human sperm quality

目的 探讨纳米细菌对人精子浓度和运动能力的影响。方法 选择2021年10~12月新乡医学院第三附属医院泌尿外科收治的12例Ⅲ型前列腺炎患者为研究对象,收集患者前列腺液标本,分离培养和鉴定前列腺液标本中纳米细菌,依据培养结果将患者分为纳米细菌阴性组和纳米细菌阳性组。应用计算机辅助精子分析系统(CASA)分析纳米细菌阴性组和纳米细菌阳性组患者的精液量、精子总活力、精子前向运动力。另取10例健康男性精液,优化处理后分为对照组和实验组;实验组精子与纳米细菌悬液共孵育,对照组精子与输卵管液共孵育;分别于孵育0、2、4、8 h时,应用CASA检测2组精子浓度、总活力、前向运动力、直线速度(VSL)、曲线速度(VCL)、平均路径速度(VAP)、精子头部侧摆幅度(ALH)和精子形态及凝集情况。结果 培养的纳米细菌为沉积在管底的白色沉淀,且培养基颜色变黄;阴性标本颜色与培养基对照组相比,颜色略有变黄。茜素红钙染色结果显示,不同视野下的纳米细菌常聚集成簇,单个细菌由于形体微小不可见,成簇的纳米细菌可被染成红色。12例Ⅲ型前列腺炎患者的前列腺液标本中,5例纳米细菌培养阳性,7例纳米细菌培养阴性。纳米细菌阴性组与纳米细菌阳性组患者的年龄、精液量、总活力比较差异无统计学意义(P>0.05),纳米细菌阳性组患者的精子浓度、前向运动力显著低于纳米细菌阴性组(P<0.001)。实验组和对照组精子形态各时间点无明显变化,均未出现明显凝集。实验组和对照组精子浓度随孵育时间的延长呈下降趋势(F=135.420、132.925,P<0.001);孵育0 h时,对照组和实验组精子浓度比较差异无统计学意义(P>0.05);孵育2、4、8 h时,实验组精子浓度显著低于对照组(P<0.001)。对照组精子总活力、前向运动力、VCL、VSL、VAP及ALH随着孵育时间的延长呈下降趋势(F=304.494、48.012、95.780、53.538、383.257、287.428,P<0.001),且实验组精子总活力、前向运动力、VCL、VSL、VAP及ALH随着孵育时间的延长亦呈下降趋势(F=1 758.057、924.174、297.114、244.244、1 173.730、290.031,P<0.001)。孵育0 h时,对照组和实验组精子总活力、前向运动力、VCL、VSL、VAP及ALH比较差异无统计学意义(P>0.05);孵育2、4、8 h时,实验组精子总活力、前向运动力、VCL、VSL、VAP及ALH显著低于对照组(P<0.05)。结论 纳米细菌可显著降低精子浓度和运动参数,进而影响患者的生育能力。

Objective To investigate the effect of nanobacteria on the concentration and motility of human sperm.Methods Twelve patients with type Ⅲ prostatitis admitted to the Department of Urology, the Third Affiliated Hospital of Xinxiang Medical University from October to December 2021 were selected as research subjects, and prostate fluid specimens from the patients were collected, the nanobacteria was isolated, cultured and identificated;then, the patients were divided into the nanobacteria-negative group and nanobacteria-positive group according the culture results, and the semen volume, total sperm motility and sperm forward motion of patients in the nanobacteria-negative group and nanobacteria-positive group were analyzed by the computer-assisted sperm analysis(CASA).The semen collected from 10 healthy male people were divided into control group and experimental group after optimization;the sperm in the experimental group was co-incubated with nanobacterial suspension, while the sperm in the control group was co-incubated with oviductal fluid;the sperm concentration, total viability, forward motility, straight line velocity(VSL),curve velocity(VCL),average path velocity(VAP),sperm head swing amplitude(ALH),sperm morphology and agglutination in the two groups were measured by CASA at 0,2,4 and 8 hours of incubation, respectively.Results The cultured nanobacteria were white precipitates deposited at the bottom of the tubes and the color of the medium turned yellow;the color of the negative specimens turned slightly yellow compared with the medium control.The results of alizarin red calcium staining showed that nanobacteria in different fields of view were often gathered into clusters and individual bacteria was too small to be visible, and clusters of nanobacteria could be stained red.Among the 12 patients with type Ⅲ prostatitis, 5 cases were positive for nanobacteria culture and 7 cases were negative for nanobacteria culture.There were no significant differences in the age, semen volume and total motility of sperm of the patients between the nanobacteria-negative group and nanobacteria-positive group(P>0.05);but the sperm concentration and forward motility of sperm of patients in the nanobacteria-positive group were significantly lower than those in the nanobacteria-negative group(P<0.001).There was no significant change in sperm morphology at all time points, and no obvious agglutination was observed in the experimental group and control group.The sperm concentration showed decreasing trend with the increase of incubation time in the experimental group and control group(F=135.420,132.925;P<0.001);At 0 hour of incubation, there was no significant difference in the sperm concentration between the experimental group and control group(P>0.05);the sperm concentration in the experimental group was significantly lower than that in the control group at 2,4,and 8 hours of incubation(P<0.001).The total sperm motility, forward motility, VCL,VSL,VAP and ALH showed decreasing trend with the increase of incubation time in the control group(F=304.494,48.012,95.780,53.538,383.257,287.428;P<0.001) and experimental group(F=1 758.057,924.174,297.114,244.244,1 173.730,290.031;P<0.001).At 0 hour of incubation, there was no significant difference in total sperm motility, forward motility, VCL,VSL,VAP and ALH between the control group and experimental group(P>0.05);at 2,4 and 8 hours of incubation, the total sperm motility, forward motility, VCL,VSL,VAP and ALH in the experimental group were significantly lower than those in the control group(P<0.05).Conclusion Nanobacteria can significantly reduce sperm concentration and motility parameters, thereby affect the fertility of patients.