Luminal fluid reabsorption plays a fundamental role in male fertility. We demonstrated that the ubiquitous GPCR signaling proteins Gq and β-arrestin-1 are essential for fluid reabsorption because they mediate coupling between an orphan receptor ADGRG2 (GPR64) and the ion channel CFTR. A reduction in protein level or deficiency of ADGRG2, Gq or β-arrestin-1 in a mouse model led to an imbalance in pH homeostasis in the efferent ductules due to decreased constitutive CFTR currents. Efferent ductule dysfunction was rescued by the specific activation of another GPCR, AGTR2. Further mechanistic analysis revealed that β-arrestin-1 acts as a scaffold for ADGRG2/CFTR complex formation in apical membranes, whereas specific residues of ADGRG2 confer coupling specificity for different G protein subtypes, this specificity is critical for male fertility. Therefore, manipulation of the signaling components of the ADGRG2-Gq/β-arrestin-1/CFTR complex by small molecules may be an effective therapeutic strategy for male infertility.

The impairment of sperm maturation is one of the major pathogenic factors in male subfertility, a serious medical and social problem affecting millions of global couples. Regrettably, the existing research on sperm maturation is slow, limited, and fragmented, largely attributable to the lack of a global molecular view. To fill the data gap, we newly established a database, namely the Sperm Maturation Database (SperMD, http://bio-add.org/SperMD ). SperMD integrates heterogeneous multi-omics data (170 transcriptomes, 91 proteomes, and five human metabolomes) to illustrate the transcriptional, translational, and metabolic manifestations during the entire lifespan of sperm maturation. These data involve almost all crucial scenarios related to sperm maturation, including the tissue components of the epididymal microenvironment, cell constituents of tissues, different pathological states, and so on. To the best of our knowledge, SperMD could be one of the limited repositories that provide focused and comprehensive information on sperm maturation. Easy-to-use web services are also implemented to enhance the experience of data retrieval and molecular comparison between humans and mice. Furthermore, the manuscript illustrates an example application demonstrated to systematically characterize novel gene functions in sperm maturation. Nevertheless, SperMD undertakes the endeavor to integrate the islanding omics data, offering a panoramic molecular view of how the spermatozoa gain full reproductive abilities. It will serve as a valuable resource for the systematic exploration of sperm maturation and for prioritizing the biomarkers and targets for precise diagnosis and therapy of male subfertility.

STUDY QUESTION:What is the mechanism behind cryoinjury in human sperm, particularly concerning the interplay between reactive oxygen species (ROS) and autophagy, and how does it subsequently affect sperm fate? SUMMARY ANSWER:The freeze-thaw operation induces oxidative stress by generating abundant ROS, which impairs sperm motility and activates autophagy, ultimately guiding the sperm toward programmed cell death such as apoptosis and necrosis, as well as triggering premature capacitation. WHAT IS KNOWN ALREADY:Both ROS-induced oxidative stress and autophagy are thought to exert an influence on the quality of frozen-thawed sperm. STUDY DESIGN, SIZE, DURATION:Overall, 84 semen specimens were collected from young healthy fertile males, with careful quality evaluation. The specimens were split into three groups to investigate the ROS-induced cryoinjury: normal control without any treatment, sperm treated with 0.5 mM hydrogen peroxide (H2O2) for 1 h, and sperm thawed following cryopreservation. Samples from 48 individuals underwent computer-assisted human sperm analysis (CASA) to evaluate sperm quality in response to the treatments. Semen samples from three donors were analyzed for changes in the sperm proteome after H2O2 treatment, and another set of samples from three donors were analyzed for changes following the freeze-thaw process. The other 30 samples were used for fluorescence-staining and western blotting. PARTICIPANTS/MATERIALS, SETTING, METHODS:Sperm motility parameters, including progressive motility (PR %) and total motility (PR + NP %), were evaluated using the CASA system on a minimum of 200 spermatozoa. The proteomic profiles were determined with label-free mass spectrometry (MS/MS) and protein identification was performed via ion search against the NCBI human database. Subsequently, comprehensive bioinformatics was applied to detect significant proteomic changes and functional enrichment. Fluorescence-staining and western blot analyses were also conducted to confirm the proteomic changes on selected key proteins. The ROS level was measured using 2',7'-dichlorodihydrofluorescein diacetate labeling and the abundance of bioactive mitochondria was determined by evaluating the inner mitochondrial membrane potential (MMP) level. Molecular behaviors of sequestosome-1 (p62 or SQSTM1) and microtubule-associated proteins 1A/1B light chain 3 (LC3) were monitored to evaluate the state of apoptosis in human sperm. Fluorescent probes oxazole yellow (YO-PRO-1) and propidium iodide (PI) were utilized to monitor programmed cell death, namely apoptosis and necrosis. Additionally, gradient concentrations of antioxidant coenzyme Q10 (CoQ10) were introduced to suppress ROS impacts on sperm. MAIN RESULTS AND THE ROLE OF CHANCE:The CASA analysis revealed a significant decrease in sperm motility for both the H2O2-treatment and freeze-thaw groups. Fluorescence staining showed that high ROS levels were produced in the treated sperm and the MMPs were largely reduced. The introduction of CoQ10 at concentrations of 20 and 30 μM resulted in a significant rescue of progressive motility (P < 0.05). The result suggested that excessive ROS could be the major cause of sperm motility impairment, likely by damaging mitochondrial energy generation. Autophagy was significantly activated in sperm when they were under oxidative stress, as evidenced by the upregulation of p62 and the increased conversion of LC3 as well as the upregulation of several autophagy-related proteins, such as charged multivesicular body protein 2a, mitochondrial import receptor subunit TOM22 homolog, and WD repeat domain phosphoinositide-interacting protein 2. Additionally, fluorescent staining indicated the occurrence of apoptosis and necrosis in both H2O2-treated sperm and post-thaw sperm. The cell death process can be suppressed when CoQ10 is introduced, which consolidates the view that ROS could be the major contributor to sperm cryoinjury. The freeze-thaw process could also initiate sperm premature capacitation, demonstrated by the prominent increase in tyrosine phosphorylated proteins, verified with anti-phosphotyrosine antibody and immunofluorescence assays. The upregulation of capacitation-related proteins, such as hyaluronidase 3 and Folate receptor alpha, supported this finding. LARGE SCALE DATA:The data underlying this article are available in the article and its online supplementary material. LIMITATIONS, REASONS FOR CAUTION:The semen samples were obtained exclusively from young, healthy, and fertile males with progressive motility exceeding 60%, which might overemphasize the positive effects while possibly neglecting the negative impacts of cryoinjury. Additionally, the H2O2 treatment conditions in this study may not precisely mimic the oxidative stress experienced by sperm after thawing from cryopreservation, potentially resulting in the omission of certain molecular alterations. WIDER IMPLICATIONS OF THE FINDINGS:This study provides substantial proteomic data for a comprehensive and deeper understanding of the impact of cryopreservation on sperm quality. It will facilitate the design of optimal protocols for utilizing cryopreserved sperm to improve applications, such as ART, and help resolve various adverse situations caused by chemotherapy, radiotherapy, and surgery. STUDY FUNDING/COMPETING INTEREST(S):This work was supported by grants from the Major Innovation Project of Research Institute of National Health Commission (#2022GJZD01-3) and the National Key R&D Program of China (#2018YFC1003600). All authors declare no competing interests. TRIAL REGISTRATION NUMBER:N/A.

Spermatogenesis is an extremely complex process, and any obstruction can cause male infertility. RhoGDIα has been identified as a risk of male sterility. In this study, we generate RhoGDIα knockout mice, and find that the males have severely low fertility. The testes from RhoGDIα-/- mice are smaller than that in WT mice. The numbers of spermatogonia and spermatocytes are decreased in RhoGDIα-/- testis. Spermatogenesis is compromised, and spermatocyte meiosis is arrested at zygotene stage in RhoGDIα-/- mice. Acrosome dysplasia is also observed in sperms of the mutant mice. At the molecular level, RhoGDIα deficiency activate the LIMK/cofilin signaling pathway, inhibiting F-actin depolymerization, impairing testis and inducing low fertility in mouse. In addition, the treatment of RhoGDIα-/- mice with Rac1 inhibitor NSC23766 alleviate testis injury and improve sperm quality by inhibiting the LIMK/cofilin/F-actin pathway during spermatogenesis. Together, these findings reveal a previously unrecognized RhoGDIα/Rac1/F-actin-dependent mechanism involved in spermatogenesis and male fertility.

Heat shock proteins (HSPs) have important roles in different developmental stages of spermatogenesis. The heat shock 70 kDa protein 5 (HSPA5) is an important component of the unfolded protein response that promotes cell survival under endoplasmic reticulum (ER) stress conditions. In this study, we explored the function of HSPA5 in spermatogenesis, by generating a germ cell-specific deletion mutant of the Hspa5 gene (conditional knockout of the Hspa5 gene, Hspa5-cKO) using CRISPR/Cas9 technology and the Cre/Loxp system. Hspa5 knockout resulted in severe germ cell loss and vacuolar degeneration of seminiferous tubules, leading to complete arrest of sper-matogenesis, testicular atrophy, and male infertility in adult mice. Furthermore, defects occurred in the sper-matogenic epithelium of Hspa5-cKO mice as early as Cre recombinase expression. Germ cell ablation of Hspa5 impaired spermatogonia proliferation and differentiation from post-natal day 7 (P7) to P10, which led to a dramatic reduction of differentiated spermatogonia, compromised meiosis, and led to impairment of testis development and the disruption of the first wave of spermatogenesis. Consistent with these results, single-cell RNA sequencing (scRNA-seq) analysis showed that germ cells, especially differentiated spermatogonia, were dramatically reduced in Hspa5-cKO testes compared with controls at P10, further confirming that HSPA5 is crucial for germ cell development. These results suggest that HSPA5 is indispensable for normal spermatogenesis and male reproduction in mice.

Spermatogenesis is an extremely complex process, and any obstruction can cause male infertility. RhoGDIα has been identified as a risk of male sterility. In this study, we generate RhoGDIα knockout mice, and find that the males have severely low fertility. The testes from RhoGDIα −/− mice are smaller than that in WT mice. The numbers of spermatogonia and spermatocytes are decreased in RhoGDIα −/− testis. Spermatogenesis is compromised, and spermatocyte meiosis is arrested at zygotene stage in RhoGDIα −/− mice. Acrosome dysplasia is also observed in sperms of the mutant mice. At the molecular level, RhoGDIα deficiency activate the LIMK/cofilin signaling pathway, inhibiting F-actin depolymerization, impairing testis and inducing low fertility in mouse. In addition, the treatment of RhoGDIα −/− mice with Rac1 inhibitor NSC23766 alleviate testis injury and improve sperm quality by inhibiting the LIMK/cofilin/F-actin pathway during spermatogenesis. Together, these findings reveal a previously unrecognized RhoGDIα/Rac1/F-actin-dependent mechanism involved in spermatogenesis and male fertility.

INTRODUCTION:Peroxiredoxin 6 (Prdx6) is widely expressed in mammalian tissues. Our previous study demonstrated that Prdx6 was expressed in human epididymis, present in human seminal fluid, and in spermatozoa. The protective role of Prdx6 in maintaining the viability and DNA integrity of human spermatozoa was also detected. Here, we demonstrate the potential role and mechanism of Prdx6 in human epididymis epithelial cells (HEECs).MATERIAL AND METHODS:Western blotting was used to measure expression levels of key proteins in the Janus kinase/signal transducer and activator of transcription (JAK/STAT) signaling pathway. The malonaldehyde (MDA) levels and antioxidant capacity in HEECs were detected with the commercial kits. Digital gene expression analysis (DGE) was used to identify gene expression patterns in control and Prdx6-interference HEECs. Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to validate the DGE findings.RESULTS:Compared to control HEECs, the expression levels of JAK1, STAT1, phosphorylated JAK1 and STAT1 were significantly increased, while the expression level of SOCS3 was significantly decreased in Prdx6-interference HEECs. The MDA level and total antioxidant capacity in Prdx6-interference HEECs were significantly increased and decreased compared to that of control, respectively. DGE analysis identified 589 up-regulated and 314 down-regulated genes (including Prdx6) in Prdx6-interference HEECs. Thirteen significantly different pathways were identified between the two groups, with the majority of genes belonging to the CCL, CXCL, IL, and IFIT family of proteins and were related to immunity. In particular, the expression levels of IL6, IL6ST, and eighteen IFN-related genes were significantly increased in Prdx6-interference HEECs compared to control HEECs.CONCLUSIONS:We found that reduced Prdx6 expression induced higher ROS levels in HEECs, which resulted in the activation of the IL-6 receptor and IFNγ expression to induce the JAK1/STAT1 signaling pathway.

Adhesion G protein-coupled receptors are elusive in terms of their structural information and ligands. Here, we solved the cryogenic-electron microscopy (cryo-EM) structure of apo-ADGRG2, an essential membrane receptor for maintaining male fertility, in complex with a Gs trimer. Whereas the formations of two kinks were determinants of the active state, identification of a potential ligand-binding pocket in ADGRG2 facilitated the screening and identification of dehydroepiandrosterone (DHEA), dehydroepiandrosterone sulfate and deoxycorticosterone as potential ligands of ADGRG2. The cryo-EM structures of DHEA-ADGRG2-Gs provided interaction details for DHEA within the seven transmembrane domains of ADGRG2. Collectively, our data provide a structural basis for the activation and signaling of ADGRG2, as well as characterization of steroid hormones as ADGRG2 ligands, which might be used as useful tools for further functional studies of the orphan ADGRG2.

系统分析了AK4在成年大鼠各主要组织(心、肝、脾、肺、肾、睾丸和附睾)及在不同周龄大鼠睾丸和附睾中的表达谱.RT-PCR及Western结果表明,AK4在各组织中呈泛在表达,且在睾丸中AK4水平从2周龄到4周龄逐渐上调,5周龄后表达趋于稳定;在附睾中,AK4在2周龄大鼠中不表达,从3周龄到4周龄表达逐渐上调,5周龄后表达基本趋于稳定.此外,构建了成年大鼠睾丸、附睾的热应激和氧化应激模型,与对照组相比,两个模型中AK4在睾丸和/或附睾组织中表达均显著上调.在应激条件下AK4的过表达对雄性生殖细胞起到一定的保护作用.本研究将为阐明雄性生殖系统中AK4在应激状态下参与的信号通路提供数据支持和理论依据.

Spermatogenesis and sperm maturation are complex and highly ordered biological processes. Any failure or disorder in these processes can cause defects in sperm morphology, motility, and fertilization ability. Cathepsin B (CTSB) is involved in the regulation of a variety of pathological processes. In the present study, we found that CTSB was abundantly expressed in the male reproductive system, however, the specific role of CTSB in regulating spermatogenesis and sperm maturation remained elusive. Hence, we generated Ctsb-/- mice using CRISPR/Cas9 technology. In Ctsb-/- mice, sperm count was significantly decreased while the level of morphologically abnormal sperm was markedly increased. Additionally, these mice had significantly lower levels of progressive motility sperm and elevated levels of immobilized sperm. Histological analysis showed slight vacuolization in the testis epithelium, as well as the loss of epididymal epithelium cells. Further investigation showed that autophagic activity was inhibited and apoptotic activity was increased in both the testis and epididymis of Ctsb-/- mice. Together, our findings demonstrate that CTSB plays an important role in spermatogenesis and sperm maturation in mice.

ALDH4A1, a member of the aldehyde dehydrogenase superfamily, is a key enzyme in the mitochondrial proline metabolism pathway. Recent studies have shown that mutations in aldh4a1 lead to reduced fertility and reproductive premature aging of male nematodes. However, the effect of ALDH4A1 on fertility of male mice has not been studied. In this study, we used CRISPR-Cas9 technology to construct a knockout mouse model of Aldh4a1 for the first time to explore the effect of this gene on the reproduction of male mice. The results showed that compared with WT male mice, Aldh4a1^(-/-) male mice were fertile, had normal spermatogenesis but defect in sperm maturation in the epididymis documented by impaired motility, increased morphological abnormalities and increased spontaneous acrosome reaction. In addition, transmission electron microscopy showed vacuoles in the sperm mitochondria, and fracture in the neck of sperms and vacuoles in these mice. These results revealed that ALDH4A1 plays a vital role in the structure of sperm flagellum and the process of sperm maturation in mice.

Spermatogenesis and sperm maturation are complex and highly ordered biological processes.Any failure or disorder in these processes can cause defects in sperm morphology, motility, and fertilization ability.Cathepsin B (CTSB) is involved in the regulation of a variety of pathological processes.In the present study, we found that CTSB was abundantly expressed in the male reproductive system, however, the specific role of CTSB in regulating spermatogenesis and sperm maturation remained elusive.Hence, we generated Ctsb -/-mice using CRISPR/Cas9 technology.In Ctsb -/-mice, sperm count was significantly decreased while the level of morphologically abnormal sperm was markedly increased.Additionally, these mice had significantly lower levels of progressive motility sperm and elevated levels of immobilized sperm.Histological analysis showed slight vacuolization in the testis epithelium, as well as the loss of epididymal epithelium cells.Further investigation showed that autophagic activity was inhibited and apoptotic activity was increased in both the testis and epididymis of Ctsb -/-mice.Together, our findings demonstrate that CTSB plays an important role in spermatogenesis and sperm maturation in mice.

I. Background: Peroxiredoxin 6 (Prdx6) is widely expressed in mammalian tissues. Our previous study demonstrated that Prdx6 was expressed in human epididymis and spermatozoa, and the protective role of Prdx6 in human spermatozoa was also reported. In this study, we demonstrate the potential role and mechanism of Prdx6 in human epididymis epithelial cells (HEECs). II. Methods and Results: Western blotting was used to measure expression levels of key proteins in the JAK / STAT signaling pathway. Digital gene expression analysis (DGE) was used to identify gene expression patterns in control HECs and in HECs after Prdx6-RNA interference (P6-RNAi). The DGE analysis identified 589 up-regulated and 314 down-regulated genes (including Prdx6) in Prdx6-RNAi (P6-RNAi) HEECs. Thirteen significantly different pathways were identified between the two groups, with the majority different expressed genes belonging to the CCL, CXCL, IL , and IFIT families. In particular, the expression levels of IL6, IL6ST, and eighteen IFN related genes were significantly increased in the condition of the down-regulated expression of Prdx6 . Compared to control HEECs, the expression levels of JAK1, STAT1, phosphorylated JAK1 and STAT1 were significantly increased, while the expression levels of SOCS3 was significantly decreased in P6-RNAi HEECs. The Malondialdehyde (MDA) level and total antioxidant capacity in P6-RNAi HEECs were significantly increased and decreased compared to that of control, respectively. III. Conclusions: We speculated that knockdown of Prdx6 resulted in higher levels of ROS in HEECs, which in turn, activated the JAK1 / STAT1 signaling pathway induced by IL-6 receptor and IFN .

Adenylate kinase 6 (AK6), a nucleus localized phosphotransferase in mammalians, shows ubiquitously expression and broad substrate activity in different tissues and cell types. Although the function of AK6 has been extensively studied in different cancer cell lines, its role in mammalian germline is still unknown. Here we showed that knockdown of AK6 inhibits cell proliferation and promotes cell apoptosis in human testicular carcinoma (NT2 cells). Co-immunoprecipitation experiment and in vitro pull down assay identified WNK1 (with no lysine kinase-1) as one of the AK6 interacting proteins in NT2 cells. Moreover, we found that AK6 regulates the phosphorylation states of WNK1 (Thr60) and affects phosphorylation level of Akt (Ser473) upon hypotonic condition, probably affecting chloride channel and regulating ion transport and homeostasis in NT2 cells and consequently contributing to the decreased cell proliferation rate. In conclusion, AK6 regulates WNK1 phosphorylation states and affects ion homeostasis in NT2 cells. These findings provide new insights into the function of AK6 and WNK1 in human testicular carcinoma. This work also provides foundation for further mechanism study of AK6 in spermatogenesis.

BACKGROUND:In the epididymis of bilateral castrated male rat, gene expression profile changed significantly. However, up to date, no study has investigated how these genes were regulated by microRNAs (miRNAs).OBJECTIVE:We investigated the alterations in the miRNA signature of the epididymis from sham-operated and bilaterally castrated rats.METHODS:By employing deep sequencing technology and qPCR, the global alterations of epididymal miRNA signature between sham-operated (Con-EP library) and bilaterally castrated rats (Cas-EP library) were explored. MiRNA-target interaction networks were annotated by GO and KEGG enrichment.RESULTS:We identified 313 and 306 known miRNAs as well as 152 and 114 novel miRNAs in the Con-EP and Cas-EP libraries, respectively. 59 miRNAs were differentially expressed, including 24 up-regulated and 35 down-regulated miRNAs, among which two up-regulated and three down-regulated ones were validated using qPCR. The expression of these miRNAs in the epididymides of rats at different postnatal ages showed regular changes from birth to adult, suggesting they were androgen-regulated. GO analysis showed that many of the miRNA targets were enriched in metabolic processes. KEGG analysis demonstrated that the targets mainly participated in the mitogen-activated protein kinase (MAPK) pathway. Moreover, 3 and 6 functional modules were detected among the up- and down-regulated miRNA target interaction networks, respectively, and these modules were involved in various biological processes.CONCLUSION:This study represents the first systematic investigation of alterations in the miRNA signature of the epididymis from bilaterally castrated rats and will provide useful resources for functional studies of the miRNAs in the male reproductive system.

INTRODUCTION High levels of reactive oxygen species (ROS) have potential toxic effects on testicular function and sperm quality. Peroxiredoxins (PRDXs) are enzymes with a role as ROS scavenger. The aim of the study was to reveal the presence and localization of PRDXs in human testis, epididymis and spermatozoa, and the protective roles of PRDX2 and PRDX6 in sperm motility. MATERIAL AND METHODS The presence and localization of PRDXs in the human testis, epididymis and spermatozoa were detected by immunohistochemistry, western blot and immunofluorescence. The effect of anti-peroxidative damage to spermatozoa was examined by adding H2O2 to the recombinant protein-treated spermatozoa. RESULTS There were strong signals of PRDX1 in spermatogonia and round spermatids; PRDX2 in the round spermatids; PRDX4 and 5 in spermatogonia; PRDX6 in Sertoli cells. PRDXs were also found in epididymal epithelial cells where the expression of PRDX1, 4, 5, 6 in the cauda was higher than in the caput of epididymis. PRDX1-6 immunoreactivity was found throughout acrosome, post-acrosomal region, equatorial segment, neck and cytoplasmic droplet, midpiece and principal piece. The H2O2-induced reduction in sperm motility was reversed by recombinant PRDX2 or PRDX6 in a dose-dependent manner. CONCLUSIONS PRDX1-6 in the human testis and epididymis presented cell-specificity. PRDX2 and 6 are potential antioxidant protectors for human spermatozoa.

42 43 Luminal fluid reabsorption plays a fundamental role in male fertility. We 44 demonstrated that the ubiquitous GPCR signaling proteins Gq and β-arrestin-1 are 45 essential for fluid reabsorption because they mediate coupling between an orphan 46 receptor ADGRG2 (GPR64) and the ion channel CFTR. A reduction in protein level 47 or deficiency of ADGRG2, Gq or β-arrestin-1 in a mouse model led to an imbalance 48 in pH homeostasis in the efferent ductules due to decreased constitutive CFTR 49 currents. Efferent ductule dysfunction was rescued by the specific activation of 50 another GPCR, AGTR2. Further mechanistic analysis revealed that β-arrestin-1 acts 51 as a scaffold for ADGRG2/CFTR complex formation in apical membranes, whereas 52 specific residues of ADGRG2 confer coupling specificity for different G protein 53 subtypes; this specificity is critical for male fertility. Therefore, manipulation of the 54 signaling components of the ADGRG2Gq/β-arrestin-1/CFTR complex by small 55 molecules may be an effective therapeutic strategy for male infertility. 56 57 Introduction 58 59 Male infertility is transforming from a personal issue to a public health problem 60 because approximately 15% of reproductive-age couples are infertile, and male 61 infertility accounts for approximately 50% of this sterility(Hamada et al., 2012 ; Jodar 62

Heat stress is demonstrated to have an effect on the function of the male testis, however, limited information has been reported on its effects on sperm maturation. In the present study, a comparative proteomic analysis was performed on the rat caput epididymal fluids responsible for sperm maturation, to identify key heat‑stress‑associated sperm maturation proteins. The results demonstrated 21 proteins corresponding to 29 differential protein spots, including 10 downregulated and 11 upregulated proteins in the heat treatment group. Functional analysis demonstrated that these proteins were primarily involved in enriched reproduction and antioxidant activity. Analysis of western blot and immunohistochemical analysis demonstrated that the expression of antioxidant proteins peroxiredoxin 6 and clusterin were downregulated, and the expression of superoxide dismutase upregulated, in the heat treatment group. Morphological and TUNEL experiments demonstrated that altered nucleus activity occurred in the caput epididymis. The study provided, to the best of our knowledge, novel information for studies on the biological functions of the epididymis and sperm maturation.

To investigate the therapeutic effect of peroxiredoxin-6(PRDX6)on ultraviolet-induced corneal injury in rats and explore the mechanism.The rat model of corneal injury was established by exposing to ultravio-let.Male wister rats were randomly divided into control groups,dexamethasone (DXM)groups and PRDX6 groups,the rats were administered four times a day and for 12 days.The corneal opacity was observed with a slit-lamp microscope.Histopathologic changes were observed with light microscopy.The content of corneal malonalde-hyde(MDA)was determined by thiobarbituric acid test and the total antioxidative capacity(TAOC)was detected by chemical colorimetric test.P38 MAPK signal pathway was detected with the method of Western blot and the gene expression of cytokines were measured by RT-PCR method.Compared with the control group,PRDX6 treat-ment significantly reduced corneal opacity,improved corneal pathology injury,decreased the MDA content and in-creased the TAOC.In the PRDX6 group the level of phosphorylated p38 protein was significantly lower than that in the control group.The gene expression of cytokine were different between control and PRDX6 groups(P <0.05).PRDX6 showed therapeutic effect in the rat model of ultraviolet-induced corneal injury.This maybe be concerned with that it could alleviated the oxidative damage,suppressed p38 MAPK phosphorylation and regulate the gene expression of cytokine.

目的 研究小鼠肾缺血再灌注后不同时间点热休克蛋白HSP90α和miRNA-144的表达变化.方法 用动脉夹夹闭小鼠双侧肾蒂45min建立肾缺血再灌注损伤模型,测定再灌注4、8、12、24、48h组及假手术组血清中肌酐(Scr)、尿素氮(BUN)水平,光镜下观察肾组织形态学改变,采用定量RT-PCR的方法检测肾缺血再灌注各时间点HSP90α mRNA和miRNA-144的表达变化,Westernblot法检测HSP90α蛋白的表达变化.结果 HE染色显示假手术组小鼠的肾组织结构及肾功能正常,再灌注组可见肾小管管腔扩张,肾小管上皮细胞有不同程度的肿胀、坏死、刷状缘消失,管腔内有管型,肾间质充血水肿,组织损伤以24h最重;肾功能指标显示再灌注组与假手术组相比,Scr、BUN水平随再灌注时间延长而升高,24h达峰值(P＜0.01).定量RT-PCR和Western blot法检测结果显示,再灌注4h后HSP90α的mRNA和蛋白表达开始逐渐增强,mRNA在12h达高峰,蛋白水平在24h达高峰(P ＜0.01);miRNA-144在再灌注后表达降低(P＜0.01).结论 HSP90α在缺血再灌注损伤的肾组织中表达增高,提示其可能参与肾缺血再灌注损伤的保护作用;miRNA-144可能为调控HSP90α功能的潜在miRNA.