AIM: To clone and express mouse peroxiredoxin Ⅰ in IEC-6 cells.METHODS: Total RNAs were isolated from cultured IEC-6 cells, and the coding region of peroxiredoxin I was amplified by RT-PCR. After it was cloned into T...AIM: To clone and express mouse peroxiredoxin Ⅰ in IEC-6 cells.METHODS: Total RNAs were isolated from cultured IEC-6 cells, and the coding region of peroxiredoxin I was amplified by RT-PCR. After it was cloned into T-vector and sequenced,pSG5 was used to transiently express peroxiredoxin I in IEC-6 by liposome-mediated transfection, and the expression of peroxiredoxin I was evaluated by RT-PCRand Western blot. RESULTS: A DNA fragment about 750 bp was amplified from total RNAs of IEC-6 cells using specific primers of peroxiredoxin Ⅰ. The sequencing confirmed the coding region was successfully cloned into T-vector, which was completely coincident with the sequence in GeneBank. After the EcoRI-BamHI fragment of T-vector containing peroxiredoxin I was inserted into pSG5, the recombinant plasmid was transferred to IEC-6 cells. RT-PCR assay showed that a DNA fragment of 930 bp could be amplified,which indicated the transcription of pSG5-Prx. Western blot confirmed the expression of peroxiredoxin Ⅰ in IEC-6 cells.CONCLUSION: Mouse peroxiredoxin I can be successfully expressed in IEC-6 cells.展开更多
This study aims to investigate the protective effects of peroxiredoxin 6 on the total motility and progressive motility of human spermatozoa.Semen samples with normal parameters were collected from 23 males and supple...This study aims to investigate the protective effects of peroxiredoxin 6 on the total motility and progressive motility of human spermatozoa.Semen samples with normal parameters were collected from 23 males and supplemented with different concentrations of peroxiredoxin 6.All the semen samples were measured according to the WHO 5th manual,and the motile spermatozoa were extracted using IVF fertilization medium supplemented with different peroxiredoxin 6 concentrations.Total motility and progressive motility were observed at different time-points of culture at room temperature.After peroxiredoxin 6 supplementation,all groups had a significant increase in total motility and progressive motility compared to the control group.The difference in total motility and progressive motility between the 0 and 10−7 mM groups was observed at 24 and 48 h of culture at room temperature.At 24 h,the total motility increased by 30%in the control group(16.03±11.91 vs.11.51±7.84),and progressive motility increased by 21%(10.53±9.4 vs.8.31±6.04).A similar trend was observed in the 48 h group.In addition,we also found that peroxiredoxin 6 had a well protective effect on sperm kinetic parameters at 10−7 mM.The findings of this study suggest that peroxiredoxin 6 can enhance sperm total motility and progressive motility in IVF fertilization medium.Peroxiredoxin 6 may have potential benefits for sperm preparation in assisted reproductive technology.展开更多
Sperm cryopreservation is useful in assisted reproductive technology and male fertility preservation.However,freezing and thawing significantly reduces the total and progressive motility of human spermatozoa.In the pr...Sperm cryopreservation is useful in assisted reproductive technology and male fertility preservation.However,freezing and thawing significantly reduces the total and progressive motility of human spermatozoa.In the present study,we explored the effects of peroxiredoxin 6(PRDX6)on total and progressive motility of human spermatozoa after cryopreservation.Semen samples of 20 males with normal parameters were collected and frozen in media supplemented with different concentrations of PRDX6(0 mM,10−5 mM,10−7 mM,and 10−9 mM,respectively).Postthaw total and progressive motility of sperms were measured.The results showed that in comparison with 0 mM,the concentrations of 10−5 mM,10−7 mM,and 10−9 mM of PRDX6 all significantly improved total motility and progressive motility of sperms(p<0.05).The 10−7 mM of PRDX6 showed the best performance.In conclusion,the supplementation of PRDX6 helps to maintain the total and progressive motility of human spermatozoa.展开更多
Objective: To study the protective effect of Peroxiredoxin 6 (PRDX6) treatment on the rat model with corneal injury caused by ultraviolet ray. Methods: SD male rats were selected as experimental animals and randomly d...Objective: To study the protective effect of Peroxiredoxin 6 (PRDX6) treatment on the rat model with corneal injury caused by ultraviolet ray. Methods: SD male rats were selected as experimental animals and randomly divided into control group, model group and PRDX6 group, corneal injury models were established by UV irradiation, and they received PRDX6 intervention. The contents of oxidative stress molecules, apoptosis molecules and MMPs/TIMPs in corneal tissues were detected on the 12 d after intervention. Results: MDA, AOPP, Fas, FasL, Bax, Caspase-3, MMP2 and MMP9 contents in corneal tissue of model group were significantly higher than those of control group while T-AOC, SOD, GSH-Px, Bcl-2, Survivin, XIAP, TIMP1 and TIMP2 contents were significantly lower than those of control group;MDA, AOPP, Fas, FasL, Bax, Caspase-3, MMP2 and MMP9 contents in corneal tissue of PRDX6 group were significantly lower than those of model group while T-AOC, SOD, GSH-Px, Bcl-2, Survivin, XIAP, TIMP1 and TIMP2 contents were significantly higher than those of model group. Conclusion: PRDX6 has inhibitory effect on the oxidative stress and apoptosis in corneal injury process caused by ultraviolet ray.展开更多
基金Supported by the National Natural Science Foundation of China,No.30230360
文摘AIM: To clone and express mouse peroxiredoxin Ⅰ in IEC-6 cells.METHODS: Total RNAs were isolated from cultured IEC-6 cells, and the coding region of peroxiredoxin I was amplified by RT-PCR. After it was cloned into T-vector and sequenced,pSG5 was used to transiently express peroxiredoxin I in IEC-6 by liposome-mediated transfection, and the expression of peroxiredoxin I was evaluated by RT-PCRand Western blot. RESULTS: A DNA fragment about 750 bp was amplified from total RNAs of IEC-6 cells using specific primers of peroxiredoxin Ⅰ. The sequencing confirmed the coding region was successfully cloned into T-vector, which was completely coincident with the sequence in GeneBank. After the EcoRI-BamHI fragment of T-vector containing peroxiredoxin I was inserted into pSG5, the recombinant plasmid was transferred to IEC-6 cells. RT-PCR assay showed that a DNA fragment of 930 bp could be amplified,which indicated the transcription of pSG5-Prx. Western blot confirmed the expression of peroxiredoxin Ⅰ in IEC-6 cells.CONCLUSION: Mouse peroxiredoxin I can be successfully expressed in IEC-6 cells.
基金supported by the Peking Post-doctoral Research Fund(EE2019-50)and Peking University International Hospital Research Funds(No.YN2019QN13).
文摘This study aims to investigate the protective effects of peroxiredoxin 6 on the total motility and progressive motility of human spermatozoa.Semen samples with normal parameters were collected from 23 males and supplemented with different concentrations of peroxiredoxin 6.All the semen samples were measured according to the WHO 5th manual,and the motile spermatozoa were extracted using IVF fertilization medium supplemented with different peroxiredoxin 6 concentrations.Total motility and progressive motility were observed at different time-points of culture at room temperature.After peroxiredoxin 6 supplementation,all groups had a significant increase in total motility and progressive motility compared to the control group.The difference in total motility and progressive motility between the 0 and 10−7 mM groups was observed at 24 and 48 h of culture at room temperature.At 24 h,the total motility increased by 30%in the control group(16.03±11.91 vs.11.51±7.84),and progressive motility increased by 21%(10.53±9.4 vs.8.31±6.04).A similar trend was observed in the 48 h group.In addition,we also found that peroxiredoxin 6 had a well protective effect on sperm kinetic parameters at 10−7 mM.The findings of this study suggest that peroxiredoxin 6 can enhance sperm total motility and progressive motility in IVF fertilization medium.Peroxiredoxin 6 may have potential benefits for sperm preparation in assisted reproductive technology.
基金Peking Post-doctoral Research Fund(EE 2019-50)Peking University International Hospital Research Funds(No.YN2019QN13)NHC Key Laboratory of Family Planning and Healthy/key laboratory of reproductive medicine of Hebei provincial(SZ-202006).
文摘Sperm cryopreservation is useful in assisted reproductive technology and male fertility preservation.However,freezing and thawing significantly reduces the total and progressive motility of human spermatozoa.In the present study,we explored the effects of peroxiredoxin 6(PRDX6)on total and progressive motility of human spermatozoa after cryopreservation.Semen samples of 20 males with normal parameters were collected and frozen in media supplemented with different concentrations of PRDX6(0 mM,10−5 mM,10−7 mM,and 10−9 mM,respectively).Postthaw total and progressive motility of sperms were measured.The results showed that in comparison with 0 mM,the concentrations of 10−5 mM,10−7 mM,and 10−9 mM of PRDX6 all significantly improved total motility and progressive motility of sperms(p<0.05).The 10−7 mM of PRDX6 showed the best performance.In conclusion,the supplementation of PRDX6 helps to maintain the total and progressive motility of human spermatozoa.
文摘Objective: To study the protective effect of Peroxiredoxin 6 (PRDX6) treatment on the rat model with corneal injury caused by ultraviolet ray. Methods: SD male rats were selected as experimental animals and randomly divided into control group, model group and PRDX6 group, corneal injury models were established by UV irradiation, and they received PRDX6 intervention. The contents of oxidative stress molecules, apoptosis molecules and MMPs/TIMPs in corneal tissues were detected on the 12 d after intervention. Results: MDA, AOPP, Fas, FasL, Bax, Caspase-3, MMP2 and MMP9 contents in corneal tissue of model group were significantly higher than those of control group while T-AOC, SOD, GSH-Px, Bcl-2, Survivin, XIAP, TIMP1 and TIMP2 contents were significantly lower than those of control group;MDA, AOPP, Fas, FasL, Bax, Caspase-3, MMP2 and MMP9 contents in corneal tissue of PRDX6 group were significantly lower than those of model group while T-AOC, SOD, GSH-Px, Bcl-2, Survivin, XIAP, TIMP1 and TIMP2 contents were significantly higher than those of model group. Conclusion: PRDX6 has inhibitory effect on the oxidative stress and apoptosis in corneal injury process caused by ultraviolet ray.
