【目的】分析谷胱甘肽过氧化物酶-5(glutathione peroxidase-5,GPx5)在正常及隐睾症双峰驼睾丸及附睾中的分布与表达,探索其在双峰驼隐睾时精子的抗氧化对生殖的调控作用。【方法】选取成年(5岁)双峰驼正常睾丸附睾10对及隐睾睾丸附睾6...【目的】分析谷胱甘肽过氧化物酶-5(glutathione peroxidase-5,GPx5)在正常及隐睾症双峰驼睾丸及附睾中的分布与表达,探索其在双峰驼隐睾时精子的抗氧化对生殖的调控作用。【方法】选取成年(5岁)双峰驼正常睾丸附睾10对及隐睾睾丸附睾6对,利用HE染色、胶原染色、网状染色观察其组织学结构特征,通过免疫组织化学染色、Western blotting、免疫荧光染色结合Image Pro Plus 13.0图像分析研究GPx5的表达及分布。【结果】与正常组相比,隐睾组双峰驼生精小管发育不良且管腔直径极显著缩小(P<0.01),间质细胞减少,附睾管腔缩小且上皮形成空泡,睾丸及附睾间质网状纤维和胶原纤维较丰富。免疫组织化学结果显示,GPx5在正常组双峰驼睾丸中未见明显表达,在隐睾组支持细胞中可见明显表达。Western blotting结果显示,与正常组相比,GPx5在隐睾组双峰驼睾丸和附睾体中表达量极显著升高(P<0.01),在附睾头中表达量极显著降低(P<0.01),在附睾尾中表达量无显著差异(P>0.05)。免疫荧光定位显示,与正常组相比,GPx5在隐睾组双峰驼睾丸和附睾尾中表达量极显著升高(P<0.01),在附睾头中表达量极显著降低(P<0.01),在附睾体中表达量无显著差异(P>0.05)。【结论】双峰驼隐睾的睾丸及附睾趋向纤维化;GPx5蛋白表达量在双峰驼隐睾睾丸中显著增加,且在支持细胞中表达增强,生精小管中的氧化应激影响了精子的正常生成;在附睾头的主细胞表达微弱,提示附睾微环境的抗氧化异常,精子损伤明显。展开更多
The mammalian epididymis not only plays a fun dame ntal role in the maturati on of spermatozoa,but also provides protecti on agai nst various stressors.The foremost among these is the threat posed by oxidative stress,...The mammalian epididymis not only plays a fun dame ntal role in the maturati on of spermatozoa,but also provides protecti on agai nst various stressors.The foremost among these is the threat posed by oxidative stress,which arises from an imbalance in reactive oxygen species and can elicit damage to cellular lipids,proteins,and nucleic acids.In mice,the risk of oxidative damage to spermatozoa is mitigated through the expression and secretion of glutathione peroxidase 5(GPX5)as a major luminal scavenger in the proximal caput epididymidal segment.Accordingly,the loss of GPX5^-/-mediated protection leads to impaired DNA integrity in the spermatozoa of aged Gpx57 mice.To explore the underlying mechanism,we have conducted transcriptomic analysis of caput epididymidal epithelial cells from aged(13 months old)Gpx5^-/-m mice.This analysis revealed the dysregulation of several thousand epididymal mRNA transcripts,in eluding the downregulation of a subgroup of piRNA pathway gen es,in aged Gpx5^-/-mice.In agreeme nt with these fin dings,we also observed the loss of piRNAs,which potentially bind to the P-element-induced wimpy testis(PlWI)-like proteins PIWIL1 and PIWIL2.The absence of these piRNAs was correlated with the elevated mRNA levels of their putative gene targets in the caput epididymidis of Gpx5^-/-mice.Importantly,the oxidative stress response genes tend to have more targeting piRNAs,and many of them were among the top increased genes upon the loss of GPX5^-/-.Taken together,our findings suggest the existence of a previously uncharacterized somatic piRNA pathway in the mammalian epididymis and its possible invoIvement in the aging and oxidative stress-mediated responses.展开更多
Tendinopathy is a common and complex musculoskeletal disorder,unfortunately current clinical strategies for tendinopathy have low therapeutic efficacy because of complicated pathogenesis.Oxidative stress is considered...Tendinopathy is a common and complex musculoskeletal disorder,unfortunately current clinical strategies for tendinopathy have low therapeutic efficacy because of complicated pathogenesis.Oxidative stress is considered as the major cause of tendinopathy as well as the important target,but still lacking ideal antioxidant solution.To this end,an efficient reactive oxygen species(ROS)biocatalyst,PtIrRuRhCu high-entropy alloy nanozyme(HEANZ),has been designed for treatment of tendinopathy.The non-ionic block copolymer(polyvinyl pyrrolidone)coated PtIrRuRhCu HEANZ with size of~4.0 nm exhibits good biocompatibility and multiple enzyme-like antioxidant activity(including peroxidase,catalase and superoxide dismutase(SOD)-like)to modulate ROS.The therapeutic efficacy of PtIrRuRhCu HEANZ in tendinopathy has been systematically demonstrated in vitro and in vivo.PtIrRuRhCu HEANZ can alleviate the t-Butyl hydroperoxide(TBHP)stimulated tendinopathy by clearing ROS,reducing inflammation and restoring mitochondrial autophagy.Using phosphoglycerate mutase family member 5(PGAM5)siRNA and FUN14 domain containing protein 1(FUNDC1)siRNA for intervention,we clearly revealed that PtIrRuRhCu HEANZ promots mitochondrial autophagy through upregulating the PGAM5/FUNDC1/glutathione peroxidase 4(GPX4)axis.This study provides a nanozyme strategy for the antioxidant treatment of tendinopathy and provides insights into the therapeutic mechanism.展开更多
文摘【目的】分析谷胱甘肽过氧化物酶-5(glutathione peroxidase-5,GPx5)在正常及隐睾症双峰驼睾丸及附睾中的分布与表达,探索其在双峰驼隐睾时精子的抗氧化对生殖的调控作用。【方法】选取成年(5岁)双峰驼正常睾丸附睾10对及隐睾睾丸附睾6对,利用HE染色、胶原染色、网状染色观察其组织学结构特征,通过免疫组织化学染色、Western blotting、免疫荧光染色结合Image Pro Plus 13.0图像分析研究GPx5的表达及分布。【结果】与正常组相比,隐睾组双峰驼生精小管发育不良且管腔直径极显著缩小(P<0.01),间质细胞减少,附睾管腔缩小且上皮形成空泡,睾丸及附睾间质网状纤维和胶原纤维较丰富。免疫组织化学结果显示,GPx5在正常组双峰驼睾丸中未见明显表达,在隐睾组支持细胞中可见明显表达。Western blotting结果显示,与正常组相比,GPx5在隐睾组双峰驼睾丸和附睾体中表达量极显著升高(P<0.01),在附睾头中表达量极显著降低(P<0.01),在附睾尾中表达量无显著差异(P>0.05)。免疫荧光定位显示,与正常组相比,GPx5在隐睾组双峰驼睾丸和附睾尾中表达量极显著升高(P<0.01),在附睾头中表达量极显著降低(P<0.01),在附睾体中表达量无显著差异(P>0.05)。【结论】双峰驼隐睾的睾丸及附睾趋向纤维化;GPx5蛋白表达量在双峰驼隐睾睾丸中显著增加,且在支持细胞中表达增强,生精小管中的氧化应激影响了精子的正常生成;在附睾头的主细胞表达微弱,提示附睾微环境的抗氧化异常,精子损伤明显。
基金This research was supported by the National Basic Research Program of China(Grant No.2014CB943103)National Natural Science Foundation of China(Grant No.31471104,No.31671203,No.31301225,No.31301226,No.31701119,and No.31571192)and was partly realized under the frame of the France-China scientific exchange programs"Xu Guangqi"and"Cai Yuanpei"of the"Partenariat Hubert CurienM attributed to J RD and YLZ The authors thank Prof.Winnie Wai Chi Shum,Prof.Xiaodong Sun,Ms.Aihua Liu,Dr.Chaobao Zhang,Dr.Zhen Lin,Dr.Xueting Luo,and the Bio-Med Big Data Center,CAS-MPG Partner Institute for Computational Biology,Shanghai Institutes for Biological Sciences,and Chinese Academy of Sciences for their kind support.
文摘The mammalian epididymis not only plays a fun dame ntal role in the maturati on of spermatozoa,but also provides protecti on agai nst various stressors.The foremost among these is the threat posed by oxidative stress,which arises from an imbalance in reactive oxygen species and can elicit damage to cellular lipids,proteins,and nucleic acids.In mice,the risk of oxidative damage to spermatozoa is mitigated through the expression and secretion of glutathione peroxidase 5(GPX5)as a major luminal scavenger in the proximal caput epididymidal segment.Accordingly,the loss of GPX5^-/-mediated protection leads to impaired DNA integrity in the spermatozoa of aged Gpx57 mice.To explore the underlying mechanism,we have conducted transcriptomic analysis of caput epididymidal epithelial cells from aged(13 months old)Gpx5^-/-m mice.This analysis revealed the dysregulation of several thousand epididymal mRNA transcripts,in eluding the downregulation of a subgroup of piRNA pathway gen es,in aged Gpx5^-/-mice.In agreeme nt with these fin dings,we also observed the loss of piRNAs,which potentially bind to the P-element-induced wimpy testis(PlWI)-like proteins PIWIL1 and PIWIL2.The absence of these piRNAs was correlated with the elevated mRNA levels of their putative gene targets in the caput epididymidis of Gpx5^-/-mice.Importantly,the oxidative stress response genes tend to have more targeting piRNAs,and many of them were among the top increased genes upon the loss of GPX5^-/-.Taken together,our findings suggest the existence of a previously uncharacterized somatic piRNA pathway in the mammalian epididymis and its possible invoIvement in the aging and oxidative stress-mediated responses.
基金supported by the National Major Research plan of NSFC(No.92368201)the National Key Research and Development Program of China(No.2021YFA1201404)+2 种基金Major Project of the National Natural Science Foundation of China(No.81991514)Jiangsu Province Medical Innovation Center of Orthopedic Surgery(No.CXZX202214)Jiangsu Provincial Key Medical Center Foundation,Jiangsu Provincial Medical Outstanding Talent Foundation,Jiangsu Provincial Medical Youth Talent Foundation and Jiangsu Provincial Key Medical Talent Foundation,and the Fundamental Research Funds for the Central Universities(Nos.14380493,and 14380494).
文摘Tendinopathy is a common and complex musculoskeletal disorder,unfortunately current clinical strategies for tendinopathy have low therapeutic efficacy because of complicated pathogenesis.Oxidative stress is considered as the major cause of tendinopathy as well as the important target,but still lacking ideal antioxidant solution.To this end,an efficient reactive oxygen species(ROS)biocatalyst,PtIrRuRhCu high-entropy alloy nanozyme(HEANZ),has been designed for treatment of tendinopathy.The non-ionic block copolymer(polyvinyl pyrrolidone)coated PtIrRuRhCu HEANZ with size of~4.0 nm exhibits good biocompatibility and multiple enzyme-like antioxidant activity(including peroxidase,catalase and superoxide dismutase(SOD)-like)to modulate ROS.The therapeutic efficacy of PtIrRuRhCu HEANZ in tendinopathy has been systematically demonstrated in vitro and in vivo.PtIrRuRhCu HEANZ can alleviate the t-Butyl hydroperoxide(TBHP)stimulated tendinopathy by clearing ROS,reducing inflammation and restoring mitochondrial autophagy.Using phosphoglycerate mutase family member 5(PGAM5)siRNA and FUN14 domain containing protein 1(FUNDC1)siRNA for intervention,we clearly revealed that PtIrRuRhCu HEANZ promots mitochondrial autophagy through upregulating the PGAM5/FUNDC1/glutathione peroxidase 4(GPX4)axis.This study provides a nanozyme strategy for the antioxidant treatment of tendinopathy and provides insights into the therapeutic mechanism.
