The Expression Of Bin1-b In Epididymis Of Oligoasthenozoospermia Rat And The Effects Of Bin1-b Antibody On Sperm

Part I The expression of Bin1-b protein and mRNA in the epididymis of oligoasthenozoospermia ratObjective To examine the expression of Bin1-b protein and its mRNA in the epididymis of oligoasthenozoospermia rat, and to study it's expression changes of pathologic epididymis.Methods Sprague-Dwaley male rats (n=30) were randomly divided into a oligoasthenozoospermia model group and a control group. The animals were treated via gavage of solution of Triptolide(10 mg/kg/d) and 1% CMC for 8 weeks, respectively. The immunohistochemical (IHC) examination and in situ hybridization (ISH) technology were performed in the epididymis of 2 groups.Results The expression of Bin1-b protein displays in the endepidermis, principal cells and basal cells of 2 groups, but the expression level of oligoasthenozoospermia group markedly increases (P<0.05). The expression of Bin1-b protein and mRNA was observed in the endepidermis, principal cells and basal cells of both groups, but the expression level of those two are higher in oligoasthenozoospermia group control one (P<0.05).Conclusion The expression of Bin1-b protein and its mRNA increases in the epididymis of oligoasthenozoospermia group. The results suggest that Bin1-b may be a endogenous defensive molecule of male reproductive system in the the pathologic state.Partâ…¡The effects of Bin1-b antibody on human spermObjective To observe the effects of Bin1-b antibody on sperm mitochondria, motility, capacitation and acrosomal reaction function in vitro, and to [0]investigate it's function on sperm of development, mature, capacitation and fertilizability in the epdidiymis.Methods Normal semen samples were selected according to the World Health Organization standards. After prepared by using a swim-up technique,, they were co-inbubation with anti-Bin1-b (final concentration of 1:200) , the blinding of Bin1-b polyclonal antibody with sperm was observed by fluorescence microscopy , the effects of Bin1-b antibody on sperm motility and capacitation were observed through chlortetracycline and Coomassie Brilliant-R250 staining, respectively. And the effects on sperm motility and mitochondrial function[0] were detected by using CASA system and JC-1 staining flow cytometry Detector.Results 1. The anti-Bin1-b polyclonal antibody and human sperm bind with sperm in the head, and non-specific binding was not observed. 2. Compared with the control group, the sperm motility decrease significantly at all time points after co-incubated with the antibody (final concentration 1:200) for 1h, 2h and 4h (P <0.05). 3. Compared with the control group, the number of sperm with high mitochondrial membrane potential reduced significantly after co-incubated with the antibody (final concentration 1:200) for 1h (P <0.05). 4. Compared with the control group, The percentage of high mitochondrial membrane potential sperm was significantly decreased after co-incubated with the antibody (final concentration 1:200) for 1h (P <0.05). 5. Compared with the control group, the percentage of acrosome reaction sperm significantly decreased after co-incubated with the antibody (final concentration 1:200) for 5h (P <0.05).Conclusion Anti-Bin1-b polyclonal antibody specific integrated with the human sperm head , a significant reduction in the mitochondrial membrane potential sperm and the sperm of athletic ability, can inhibit the sperm and the acrosome reaction, and other closely related to the reproductive physiology function.