| Objectives Deficiency of trace element zinc in seminal plasma is one of the important causes of low sperm motility.This research aims to explore mechanism by which zinc supplementation in semen improves sperm motility and to explore the role of G proteincoupled receptor 39(GPR39)in the effect of trace element zinc on sperm motility.Methods 40 semen samples including 20 cases of asthenospermia(PR+NR<40% and PR<32%)and 20 cases of normal semen(PR+NR≥40% and PR≥32%)which met the requirement were collected from Xingtai Infertility Specialized Hospital from July 2021 to December 2021 entered research.The semen was divided into 2 parts,one of which was added with zinc(50μg/m L),and the other was not treated.Sperm parameters measured by CASA after 1 hour.According to the changes of sperm motility after the zinc supplementation experiment,asthenospermia semen were divided into asthenospermia increased motility group(AP group,12)and asthenospermia motility unaffected group(AN group,8).Normal semen was divided into normal motility increased group(CP group,10).and normal vitality unaffected group(CN group,10).We selected 6 cases randomly from each group for follow-up experiments.Detection of zinc level in seminal plasma by trace element analyzer,detection of free zinc expression in sperm by Zinquin Ethyl Ester combined with confocal laser,observation of GPR39 expression in Sperm by Immunofluorescence Staining,detection of m RNA expression of GPR39,AQP3,AQP7,MT1 and MT2 in sperm by q PCR,detection of SOD and MDA in Seminal Plasma by ELISA.Results 1 There was no significant difference in seminal plasma zinc levels between normal semen and asthenospermia semen before zinc supplementation(P>0.05),but zinc levels in seminal plasma increased significantly after zinc supplementation(P<0.05).2 The fluorescence of free zinc in asthenospermia spermatozoa was enhanced in the sperm head and sperm tail after zinc supplementation,but there is no obvious change in normal sperm.3 Comparing the m RNA expressions of 9 zinc transporters and 10 zinc-iron transporters in the zinc transporter family,there was no significant change in the expression of zinc transporters after zinc supplementation(P>0.05).4 GPR39 was widely expressed in sperm and was mainly located in the sperm head and the mid-neck of sperm tail.5 Compared with no zinc supplementation,the m RNA expressions of GPR39 and AQP7 in sperm of the CP group increased after zinc supplementation(P<0.05),and the level of MT1 in seminal plasma increased in normal semen(P<0.05).6 Compared with no zinc supplementation,after zinc supplementation,the m RNA expressions of GPR39 and AQP7 in sperm of AP group increased,the level of SOD in seminal plasma increased(P<0.05),and the m RNA expression of AQP3 and AQP7 in sperm of AN group increased in asthenospermia semen(P<0.05).7 Compared with no zinc supplementation,there was no significant difference in the m RNA expression of MT2 in sperm and the level of MDA in seminal plasma between the groups after zinc supplementation(P>0.05).Conclusions GPR39,AQP7,MT1 and SOD play important roles on improving sperm motility after zinc supplementation,but the mechanisms of improving function of normal sperm and weak sperm are different.In normal sperm,zinc promote the expression of MT1 mainly through GPR39-mediated enhancement of AQP7 to improve sperm motility.In asthenospermia,GPR39-mediated AQP7 regulates the content of SOD to improve sperm motility.Figure 11;Table 6;Reference 130... |
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