Testis Tissue Research Articles

Dapagliflozin ameliorates diabetes-induced spermatogenic dysfunction by modulating the adenosine metabolism along the gut microbiota-testis axis

Male infertility is one of the most common complications of diabetes mellitus (DM). Dapagliflozin is widely used to manage the type II DM. This study aimed to assess the dapagliflozin’s effects on the spermatogenesis by administering either dapagliflozin (Dapa) or vehicle (db) to male db/db mice, and using littermate male db/m mice as the control (Con). We further performed the integrative analyses of the cecal shotgun metagenomics, cecal/plasmatic/testicular metabolomics, and testicular proteomics. We found that dapagliflozin treatment significantly alleviated the diabetes-induced spermatogenic dysfunction by improving sperm quality, including the sperm concentration and sperm motility. The overall microbial composition was reshaped in Dapa mice and 13 species (such as Lachnospiraceae bacterium 3–1) were regarded as potential beneficial bacteria. Metabolites exhibited modified profiles, in which adenosine, cAMP, and 2′-deoxyinosine being notably altered in the cecum, plasma, and testis, respectively. Testicular protein expression patterns were similar between the Dapa and Con mice. In vivo results indicated that when compared with db group, dapagliflozin treatment alleviated apoptosis and oxidative stress in testis tissues by down-regulating 2′-deoxyinosine. This was further validated by in vitro experiments using GC-2 cells. Our findings support the potential use of dapagliflozin to prevent the diabetes-induced impaired sperm quality and to treat diabetic male infertility.

Molecular and cellular regulation on sexual fate and gonadal development in hermaphrodite yellowfin seabream Acanthopagrus latus

Yellowfin seabream (Acanthopagrus latus) is a hermaphroditic fish with male prematurity, which is also an economically important mariculture species in coastal areas of China. Although the production yield has been increasing in recent years, artificial breeding of this species remains imperfect due to the lack understanding of its sex differentiation mechanism. In this study, we conducted transcriptomic sequencing to screen for sex-related genes in different stage of the testis, ovotestis, ovary, hypothalamus, and pituitary tissues of yellowfin seabream. During gonad development (from intersex to mature stage), Gnrh3, Kiss2, and Dbh were expressed in the hypothalamus and Lhb and Fshb were expressed in the pituitary. Sex-related genes such as Hsd11b1, Foxl2l, Amh, and Gsdf were among the top 10 downregulated genes in the intersex testis compared with the mature testis. The expression of Hsd11b, Wnt, and Sox families genes differed significantly in the sex change process among functional mature gonads. In addition, treatment with estradiol resulted in feminizing of the ovotestis. Inhibition of androgens or estrogens promoted ovarian or testicular steps of ovotestis development. In summary, we systematically evaluated gene expression profiles during yellowfin seabream gonad development from intersex to functional male or female. Our results confirmed that regulation by the hypothalamus-pituitary-gonad (HPG) axis and the promotion of sex steroid hormones acted on this process, and they provide a reference for further studies of the involvement of the HPG axis in sex change and of sex induction methods in yellowfin seabream.

Analysis of the differentially expressed genes in the combs and testes of Qingyuan partridge roosters at different developmental stages

BackgroundThe sexual maturity of chickens is an important economic trait, and the breeding of precocious and delayed puberty roosters is an important selection strategy for broilers. The comb serves as an important secondary sexual characteristic of roosters and determines their sexual precocity. Moreover, comb development is closely associated with gonad development in roosters. However, the underlying molecular mechanism regulating the sexual maturity of roosters has not yet been fully explored.ResultsIn order to identify the genes related to precocious puberty in Qingyuan partridge roosters, and based on the synchrony of testis and combs development, combined with histological observation and RNA-seq method, the developmental status and gene expression profile of combs and testis were obtained. The results showed that during the early growth and development period (77 days of age), the development of combs and testis was significant in the high comb (H) group versus the low comb (L) group (p < 0.05); however, the morphological characteristic of the comb and testicular tissues converged during the late growth and development period (112 days of age) in the H and L groups. Based on these results, RNA-sequencing analysis was performed on the comb and testis tissues of the 77 and 112 days old Qingyuan Partridge roosters with different comb height traits. GO and KEGG analysis enrichment analysis showed that the differentially expressed genes were primarily enriched in MAPK signaling, VEGF signaling, and retinol metabolism pathways. Moreover, weighted correlation network analysis and module co-expression network analysis identified WNT6, AMH, IHH, STT3A, PEX16, KPNA7, CATHL2, ROR2, PAMR1, WISP2, IL17REL, NDRG4, CYP26B1, and CRHBP as the key genes associated with the regulation of precocity and delayed puberty in Qingyuan Partridge roosters.ConclusionsIn summary, we identified the key regulatory genes of sexual precocity in roosters, which provide a theoretical basis for understanding the developmental differences between precocious and delayed puberty in roosters.

The protective role of ascorbic acid on the testis tissue damage induced by paracetamol in albino rats

Ascorbic acid (AA) plays roles in many biological functions, such as participating in the production of collagen by taking a role in proline and lysine hydroxylation. AA works by removing the reactive oxygen species, thus removing the adhesion of neutrophils to endothelium. The present study investigates the protective effects of AA on the testis tissue damage induced by paracetamol in rats. Thirty-two male rats were equally divided into four groups, with eight rats in each. Group (1) regular control group, group (2) was received 500 mg/kg/b.w. of AA orally for two weeks, group (3) was received 500 mg/kg/b.w. of paracetamol orally for two weeks, group (4) was treated with the AA (500 mg/kg/b.w.) and paracetamol (500 mg/kg/b.w.) orally for two weeks. The histopathological investigations of the testis tissues from the paracetamol group showed atrophy and degeneration of seminiferous tubules with an absence of spermatozoa, presence of cell debris, and loss of the Sertoli cells in some seminiferous tubules when compared with control animals. The testis tissues from a protective group showed less damage in the tubules and germ cells when compared with the paracetamol group. This study indicates that AA protects against paracetamol-induced testis damage in adult male rats.

MRNA-Seq of testis and liver tissues reveals a testis-specific gene and alternative splicing associated with hybrid male sterility in dzo.

Alternative splicing (AS) plays an important role in the co-transcription and post-transcriptional regulation of gene expression during mammalian spermatogenesis. The dzo is the male F1 offspring of an interspecific hybrid between a domestic bull (Bos taurus ♂) and a yak (Bos grunniens ♀) which exhibits male sterility. This study aimed to identify the testis-specific genes and AS associated with hybrid male sterility in dzo. The iDEP90 program and rMATS software were used to identify the differentially expressed genes (DEG) and differential alternative splicing genes (DSG) based on RNA-seq data from the liver (n = 9) and testis (n = 6) tissues of domestic cattle, yak, and dzo. Splicing factors (SF) were obtained from the AmiGO2 and the NCBI databases, and Pearson correlation analysis was performed on the differentially expressed SFs and DSGs. We focused on the testis-specific DEGs and DSGs between dzo and cattle and yak. Among the top 3,000 genes with the most significant variations between these 15 samples, a large number of genes showed testis-specific expression involved with spermatogenesis. Cluster analysis showed that the expression levels of these testis-specific genes were dysregulated during mitosis with a burst downregulation during the pachynema spermatocyte stage. The occurrence of AS events in the testis was about 2.5 fold greater than in the liver, with exon skipping being the major AS event (81.89% to 82.73%). A total of 74 DSGs were specifically expressed in the testis and were significantly enriched during meiosis I, synapsis, and in the piRNA biosynthesis pathways. Notably, STAG3 and DDX4 were of the exon skipping type, and DMC1 was a mutually exclusive exon. A total of 36 SFs were significantly different in dzo testis, compared with cattle and yak. DDX4, SUGP1, and EFTUD2 were potential SFs leading to abnormal AS of testis-specific genes in dzo. These results show that AS of testis-specific genes can affect synapsis and the piRNA biosynthetic processes in dzo, which may be important factors associated with hybrid male sterility in dzo.

Protective effect of baicalin on cyclophosphamideinduced oxidative stress and morphologic damage in testis tissue

Protective effect of baicalin on cyclophosphamideinduced oxidative stress and morphologic damage in testis tissue

NAD+ precursors promote the restoration of spermatogenesis in busulfan-treated mice through inhibiting Sirt2-regulated ferroptosis.

Rationale: In recent years, nicotinamide adenine dinucleotide (NAD+) precursors (Npre) have been widely employed to ameliorate female reproductive problems in both humans and animal models. However, whether and how Npre plays a role in the male reproductive disorder has not been fully clarified. Methods: In the present study, a busulfan-induced non-obstructive azoospermic mouse model was used, and Npre was administered for five weeks following the drug injection, with the objective of reinstating spermatogenesis and fertility. Initially, we assessed the NAD+ level, germ cell types, semen parameters and sperm fertilization capability. Subsequently, testis tissues were examined through RNA sequencing analysis, ELISA, H&E, immunofluorescence, quantitative real-time PCR, and Western blotting techniques. Results: The results indicated that Npre restored normal level of NAD+ in blood and significantly alleviated the deleterious effects of busulfan (BU) on spermatogenesis, thereby partially reestablishing fertilization capacity. Transcriptome analysis, along with recovery of testicular Fe2+, GSH, NADPH, and MDA levels, impaired by BU, and the fact that Fer-1, an inhibitor of ferroptosis, restored spermatogenesis and semen parameters close to CTRL values, supported such possibility. Interestingly, the reduction in SIRT2 protein level by the specific inhibitor AGK2 attenuated the beneficial effects of Npre on spermatogenesis and ferroptosis by affecting PGC-1α and ACLY protein levels, thus suggesting how these compounds might confer spermatogenesis protection. Conclusion: Collectively, these findings indicate that NAD+ protects spermatogenesis against ferroptosis, probably through SIRT2 dependent mechanisms. This underscores the considerable potential of Npre supplementation as a feasible strategy for preserving or restoring spermatogenesis in specific conditions of male infertility and as adjuvant therapy to preserve male fertility in cancer patients receiving sterilizing treatments.

Comparative Evaluation of Native and Liposomal Curcumin Against Acute Reproductive Toxicity Induced by Cadmium Chloride in Male Mice

Exposure to cadmium chloride (Cd) is linked to reproductive disorders. Curcumin (CUR), a traditional medicinal compound, has been considered for its therapeutic properties. The present study evaluated the antioxidant effects of CUR and liposomal CUR on Cd‐induced testicular toxicity. Mice were exposed to Cd (2.5 mg/kg as a single intraperitoneal dose), Cd + CUR at 50, 100, and 200 mg/kg, and Cd + liposomal CUR (15 mg/kg) orally for 7 days. At the end of the treatment period, histological studies, sperm parameters, oxidative stress parameters, and blood–testis barrier (BTB) integrity were performed. Cd‐induced histopathological deleterious changes in the testis and epididymis tissues increased BTB permeability. It caused a significant decrease in sperm count and viability (p &lt; 0.001), as well as an increase in sperm abnormality (p &lt; 0.001) and a reduction in the antioxidant activity of superoxide dismutase (SOD), glutathione (GSH) (p &lt; 0.001), and increased lipid peroxidation (LPO; p &lt; 0.001) compared to the control group. Co‐treatment of CUR with Cd dose‐dependently and significantly attenuated Cd’s destructive effects on sperm and oxidative stress parameters at 50 mg/kg (p &lt; 0.01) and 100, 200 mg/kg (p &lt; 0.001). Although the liposomal form of CUR showed protective effects against some Cd‐induced changes, such as sperm viability and count (p &lt; 0.001), it did not significantly improve LPO and GSH levels. The results indicated that native CUR prevented Cd‐induced reproductive toxicity in male mice by suppressing oxidative injury, and there was no superior effect of liposomal CUR over native CUR.

ARTS is essential for di-2-ethylhexyl phthalate (DEHP)-induced apoptosis of mouse Leydig cells

As an extensively employed plasticizer in industrial applications, di-2-ethylhexyl phthalate (DEHP) can induce apoptosis of mouse Leydig cells, yet the precise mechanism remains elusive. In the current study, we identified that DEHP could specially induced apoptosis in the Leydig cells of the testis tissue, accompanied with the upregulation of apoptosis-related protein in the TGF-β signaling pathway (ARTS) in the cells. Overexpression of ARTS significantly induced apoptosis of TM3 cells, while knockdown of ARTS inhibited apoptosis. Furthermore, DEHP-induced apoptosis of TM3 cells could be alleviated by knockdown of ARTS, which indicated that ARTS was involved in DEHP-induced apoptosis of mouse Leydig cells. Bioinformation assay predicts that there are four potential p53-responsive elements (p53-REs) located at − 6060, − 5726, − 5631 and − 5554 before the transcription start site of ARTS gene, implying that gene transcription of ARTS could be regulated by p53. Interestingly, DEHP was shown to specifically upregulate the expression of p53 in the Leydig cells of the testis tissue and TM3 cells. Consistently, p53 was proved to bind to the RE4 site of the ARTS gene promoter and transcriptionally activated the promoter-driven expression of the luciferase reporter gene. Overexpression of p53 could induce apoptosis of TM3 cells; while knockdown of p53 could not only rescue DEHP-induced apoptosis of the cells, but also inhibit DEHP-caused upregulation of ARTS. Meanwhile, we showed that oxidative stress could induce apoptosis of TM3 cells, accompanied with the increased protein levels of p53 and ARTS; while inhibition of oxidative stress dramatically alleviated DEHP-induced apoptosis and the up-regulation of p53 and ARTS. Taken together, these results indicated that DEHP-induced oxidative stress activates the p53-ARTS cascade to promote apoptosis of mouse Leydig cells.

Identification and validation of SHC1 and FGFR1 as novel immune-related oxidative stress biomarkers of non-obstructive azoospermia.

Non-obstructive azoospermia (NOA) is a major contributor of male infertility. Herein, we used existing datasets to identify novel biomarkers for the diagnosis and prognosis of NOA, which could have great significance in the field of male infertility. NOA datasets were obtained from the Gene Expression Omnibus (GEO) database. CIBERSORT was utilized to analyze the distributions of 22 immune cell populations. Hub genes were identified by applying weighted gene co-expression network analysis (WGCNA), machine learning methods, and protein-protein interaction (PPI) network analysis. The expression of hub genes was verified in external datasets and was assessed by receiver operating characteristic (ROC) curve analysis. Gene set enrichment analysis (GSEA) was applied to explore the important functions and pathways of hub genes. The mRNA-microRNA (miRNA)-transcription factors (TFs) regulatory network and potential drugs were predicted based on hub genes. Single-cell RNA sequencing data from the testes of patients with NOA were applied for analyzing the distribution of hub genes in single-cell clusters. Furthermore, testis tissue samples were obtained from patients with NOA and obstructive azoospermia (OA) who underwent testicular biopsy. RT-PCR and Western blot were used to validate hub gene expression. Two immune-related oxidative stress hub genes (SHC1 and FGFR1) were identified. Both hub genes were highly expressed in NOA samples compared to control samples. ROC curve analysis showed a remarkable prediction ability (AUCs > 0.8). GSEA revealed that hub genes were predominantly enriched in toll-like receptor and Wnt signaling pathways. A total of 24 TFs, 82 miRNAs, and 111 potential drugs were predicted based on two hub genes. Single-cell RNA sequencing data in NOA patients indicated that SHC1 and FGFR1 were highly expressed in endothelial cells and Leydig cells, respectively. RT-PCR and Western blot results showed that mRNA and protein levels of both hub genes were significantly upregulated in NOA testis tissue samples, which agree with the findings from analysis of the microarray data. It appears that SHC1 and FGFR1 could be significant immune-related oxidative stress biomarkers for detecting and managing patients with NOA. Our findings provide a novel viewpoint for illustrating potential pathogenesis in men suffering from infertility.

Resveratrol Supplementation is Associated with Elemental Metabolism in Some Tissues of Acute Swimming Rats

The aim of this study was to investigate how resveratrol administration affects element metabolism in some tissues in rats undergoing acute swimming exercise. In the study, 28 adult male Wistar rats were used and the animals were divided into 4 groups in equal numbers. Group 1, Control; Group 2, Swimming: The group fed a standard diet and had 30 minutes of acute swimming exercise. Group 3, Resveratrol: The group receiving 10 mg/kg resveratrol supplement for four weeks. Group 4, Resveratrol + Swimming: The group that received 10 mg/kg resveratrol supplement for four weeks and also had 30 minutes of acute swimming exercise. At the end of the four-weeks diet, group 2 and group 4 animals were given 30 minutes of acute swimming exercise. Animals were sacrificed immediately after swimming practices, and testis, kidney, heart and liver tissue samples were taken. Elemental analyzes in tissue samples were determined by atomic emission method. Elements measured in testicular tissue were not affected by exercise and resveratrol applications. Both resveratrol administration and acute swimming exercise resulted in changes in zinc, iron, magnesium, selenium, molybdenum levels in kidney tissue, iron and calcium levels in heart tissue, and zinc, iron and molybdenum levels in liver tissue (p&amp;lt;0.05). The results of study, which revealed that resveratrol administration caused changes in element metabolism in some tissues of rats both during and independently of exercise, can be presented as the first and original finding based on med-line scans.

Effects of Immunocastration and Amino Acid Supplementation on Yearling Fallow Deer (Dama dama) Testes Development.

Forty-four fallow deer bucks (10 months old; 22.9 ± 2.4 kg) were utilized to investigate the effects of immunocastration and amino acid supplementation on testes development. Immunocastrated bucks were administered Improvac® at weeks 1, 8, and 20 of this study (control group: intact males). Starting at week 8, half of each sex received rumen-protected lysine and methionine (3:1) supplementation. At slaughter (week 37/39), body size, internal fat deposits, antler size parameters, testes weight, testes surface color, cauda epididymal sperm viability and morphology, and seminiferous tubule circumference and epithelium thickness were determined. Animals with larger body sizes, greater forequarter development, and antler growth also had greater testes development. Whilst the result of immunocastration on testes size is unexpected, testes tissue showed impaired development (atrophied seminiferous tubules), decreased sperm viability, and normal morphology. Testes tissue from immunocastrated deer was less red, possibly indicating reduced blood supply. Conversely, amino acid supplementation increased testes' redness and sperm viability, and intact males fed amino acids showed the greatest seminiferous tubule development. Thus, immunocastration may be a welfare-friendly alternative for venison production. Whilst the results support findings from the literature that testes size is not a reliable indicator of immunocastration success, this warrants further investigation in deer over different physiological development stages.

Exploring Intercellular Dynamics: Ultra-Weak Biophoton Emission as a Novel Indicator of Altered Cell Functions and Disease in Oligospermia Mice.

Introduction: Biophoton emission, the spontaneous release of photons from living cells, has emerged as an attractive field of research in the study of biological systems. Scientists have recently discovered that changes in biophoton emission could serve as potential indicators of pathological conditions. This intriguing phenomenon suggests that cells might communicate and interact with each other through the exchange of these faint but significant light signals. Therefore, the present study introduces intercellular relationships with biophoton release to detect normal and abnormal cell functions to further achieve cellular interactions by focusing on cell and cell arrangement in disease conditions. Methods: Twenty male mice were assigned to control and busulfan groups. Five weeks after the injection of busulfan, the testis was removed, and then the stereological techniques and TUNEL assay were applied to estimate the histopathology of the testis tissue sections. Results: The findings revealed that the ultra-weak biophoton emission in the control group was significantly lower than in the busulfan group. The oligospermia mice model showed that it significantly changed the spatial arrangement of testicular cells and notably decreased the testis volume, length of seminiferous tubules, and the number of testicular cells. The results of the TUNEL assay showed that the percentage of apoptotic cells significantly increased in the busulfan group. Conclusion: The ultra-weak biophoton emission from testis tissue was reduced in oligospermia mice. As a result, the decline of ultra-weak biophoton can indicate a change in cell arrangement, a decrease in intercellular interaction, and eventually disease.

Testicular piRNA Analysis Identified Dysregulated piRNAs in Non-obstructive Azoospermia.

Male infertility has remained idiopathic in a remarkable proportion of all cases. Gonadal expression of PIWI-interacting RNAs (piRNAs) has been shown to be vital to normal spermatogenesis, as they are expressed in almost all types of testicular germ cells. These molecules and their related Piwi proteins strictly regulate transposable elements' activity and gene expression. We aimed to identify dysregulated piRNAs in idiopathic non-obstructive azoospermic (NOA) testis by global expression analysis. Testis tissue samples from 18 azoospermic patients (ten NOA and eight OA) were studied by small RNA sequencing. To validate high-throughput sequencing data, quantitative real-time polymerase chain reactions for two differentially altered piRNAs were performed. Bioinformatics analyses were undertaken to identify pathways affected by piRNA dysregulation. In the NOA group, 1328 piRNAs were identified to be differentially expressed, of which 1322 were downregulated and 6 were upregulated. Bioinformatics analysis corroborated the involvement of dysregulated piRNA in spermatogenesis. We also identified 64 clusters of differentially expressed piRNAs, of which 42 clusters had a minimum of ten absolute piRNA hits. Our study suggests that piRNAs show significant dysregulation in infertility. Their target genes play a role in their self-biogenesis, probably by regulating their own production through a feedback mechanism. The downregulated piRNAs may find value as biomarkers for the presence of spermatozoa in the testis of azoospermic individuals, while the upregulated piRNAs are great candidates for further investigation of their precise functions in spermatogenesis.

Impact of Z chromosome inversions on gene expression in testis and liver tissues in the zebra finch.

Chromosomal inversions have been identified in many natural populations and can be responsible for novel traits and rapid adaptation. In zebra finch, a large region on the Z chromosome has been subject to multiple inversions, which have pleiotropic effects on multiple traits but especially on sperm phenotypes, such as midpiece and flagellum length. To understand the effect, the Z inversion has on these traits, we examined testis and liver transcriptomes of young males at different maturation times. We compared gene expression differences among three inversion karyotypes: AA, B*B* and AB*, where B* denotes the inverted regions on Z with respect to A. In testis, 794 differentially expressed genes were found and most of them were located on chromosome Z. They were functionally enriched for sperm-related traits. We also identified clusters of co-expressed genes that matched with the inversion-related sperm phenotypes. In liver, there were some enriched functions and some overrepresentation on chromosome Z with similar location as in testis. In both tissues, the overrepresented genes were located near the distal end of Z but also in the middle of the chromosome. For the heterokaryotype, we observed several genes with one allele being dominantly expressed, similar to expression patterns in one or the other homokaryotype. This was confirmed with SNPs for three genes, and interestingly one gene, DMGDH, had allele-specific expression originating mainly from one inversion haplotype in the testis, yet both inversion haplotypes were expressed equally in the liver. This karyotype-specific difference in tissue-specific expression suggests a pleiotropic effect of the inversion and thus suggests a mechanism for divergent phenotypic effects resulting from an inversion.

A Comparative Transcriptomic Study and Key Gene Targeting of Lamprey Gonadal Immune Response

ABSTRACT The mammalian testis and ovary possess special immunocompetence, which is central to provide protection against pathogens. However, the innate immune responses to immune challenges in lamprey gonads are poorly understood. In this study, we extracted RNA from testis and ovary tissues of lampreys at 0 hour, 8 hours and 17 days after lipopolysaccharides (LPS) stimulation and performed transcriptome sequencing. While the transcriptome profiles of the two tissues were different for the most part, genes LIP, LECT2, LAL2, GRN, ITLN, and C1q were found to be the most significantly up-regulated genes in both. Quantitative Real-time PCR (qRT-PCR) analysis confirmed that these genes were upregulated after stimulation. Furthermore, immunohistochemical staining showed that these genes in lamprey gonads are expressed in high quantities and have a specific distribution. Taken together, our results suggest that these genes could play an essential role in response of the gonads to LPS induction. This research establishes a basis for investigating the immune mechanism of vertebrate gonads and presents a fresh concept for gaining insight into the evolutionary development of jawless vertebrates.

Defensive Role of Vitamin E Against Nitrosamine-Induced Harm to Testicular Tissues in Male BALB/C Albino Mice

Nitrosamines are toxic substances that induce oxidative stress. Oxidative stress results from an imbalance between the formation of free radicals (ROS) and their removal from the body and is a direct or indirect cause of a variety of diseases. The research aimed to assess vitamin E's potential protective role against nitrosamine-induced defects of testicular tissues. A total of forty Albino mice, weighing between 20 To25 grams and aged 25 to 30 weeks, were employed for the study. These mice are divided into five groups, each consisting of eight individuals. The initial group served as control, second group received a daily oral dosage of 0.05 mL/ g body weight of nitrosamine over one month. The third group was orally administered 0.1 mL/g of vitamin E. Groups IV and V were provided with vitamin E doses of 0.1 and 0.2 mL/g body weight orally Daily for a month, followed by a nitrosamine dose of 0.05 mL/g. Post the treatment period, blood samples were obtained, and the testis and epididymis tissues were subjected to examination. The study findings revealed notable alterations, encompassing the inhibition of spermatogenesis, reduction in diameters of seminiferous tubules within the testis and epididymis, as well as epithelial cell thickness. Additionally, severe necrosis and germ cell erosion were observed. In contrast, results from the vitamin E-treated animals exhibited favorable changes in the testicular and epididymis tissues when compared to the nitrosamine-exposed group and the control group. Moreover, histological assessments of animals administered a preventive vitamin E dose (0.2 mL/g) alongside nitrosamine indicated enhanced spermatogenesis, evidenced by augmented seminiferous tubule diameters and increased epithelial cell thickness within the testis and epididymis.

Salvia officinalis restores semen quality and testicular functionality in cadmium-intoxicated male rats

The present study investigated the potential ability of Salvia officinalis, one of the oldest medicinal plants, to protect male rats against cadmium reproductive toxicity. Twenty-eight healthy male rats were randomly allocated into four groups (n = 7); control, Salvia-extract treated group, cadmium treated group and a group treated with both Cd and Salvia. Administration of cadmium reduced the relative testis to body weight and significantly affected sperm parameters by decreasing motility, viability, count and increasing morphological aberrations. Comet assay was used to detect DNA fragmentation in sperms of the rats exposed to Cd. Serum levels of testosterone T, follicle stimulating hormone FSH, and luteinizing hormone LH were significantly decreased. The biochemical analysis of testicular tissue showed a significant rise in Malondialdehyde MDA level coupled with a decrease in the activity of antioxidant enzymes (superoxide dismutase SOD, glutathione peroxidase GPx and catalase CAT). The histological examination of testis sections after Cd administration revealed severe degeneration of spermatogenic cells. Seminiferous tubules were filled with homogenous eosinophilic fluid associated with atrophy of other seminiferous tubules. Co-treatment with the Salvia officinalis extract restored the oxidative enzymes activities and decreased the formation of lipid peroxidation byproduct, which in turn ameliorated the effect of Cd on sperm parameters, DNA damage and testis histopathology. Taken together, it can be concluded that the synergistic antioxidant and radical savaging activities of Salvia officinalis prevented the effect of Cd on semen quality, sperm DNA damage, along with the oxidative stress and histological abnormalities in the testis tissues.

Localization and expression of phospholipase A2 and polyunsaturated fatty acid profile in the testis tissues of Hu sheep

The fatty acid content and the localization and expression of phospholipase A2 (PLA2) in the testis of Hu sheep were investigated. A total of 18 six-month-old Hu sheep were divided into small group (S, with left testis weight < 50 g), medium group (M, with left testis weight among 90–110 g), and large group (L, with left testis weight >160 g), which had six individuals each. The expression of PLA2 in testicular tissues of different sizes was analyzed by immunohistochemistry, RT-qPCR, and Western blot. The fatty acid profile was detected by gas chromatography. Immunohistochemical labeling determined that PLA2 protein was expressed in the Leydig and Sertoli cells of testis, and the immunohistochemical average optional density in the S group was significantly greater than the L group (P < 0.05). RT-qPCR and Western blot analysis showed that PLA2 in the S group was greater than that in the L group (P < 0.05). Docosahexaenoic acid, ω-3 polyunsaturated fatty acid (PUFA), and total PUFA content in the testis of the L group were significantly less than those of the S and M groups (P < 0.01). This study showed that PLA2 content in the S group was greater than that in the L group.

Selenium reduces acrylamide-induced testicular toxicity in rats by regulating HSD17B1, StAR, and CYP17A1 expression, oxidative stress, inflammation, apoptosis, autophagy, and DNA damage.

This study investigated the effects of Selenium (Se) on testis toxicity induced by Acrylamide (ACR) in rats. In our study, 50 male adult rats were used, and the rats were divided into five groups; control, ACR, Se0.5 + ACR, Se1 + ACR, and Se1. Se and ACR treatments were applied for 10 days. On the 11th day of the experimental study, intracardiac blood samples from the rats were taken under anesthesia and euthanized. Sperm motility and morphology were evaluated. Dihydrotestosterone, FSH, and LH levels in sera were analyzed with commercial ELISA kits. MDA, GSH, TNF-α, IL-6, and IL-1β levels and SOD, GPx, and CAT, activities were measured to detect the level of oxidative stress and inflammation in rat testis tissues. Expression analysis of HSD17B1, StAR, CYP17A1, MAPk14, and P-53 as target mRNA levels were performed with Real Time-PCR System technology for each cDNA sample synthesized from rat testis RNA. Testicular tissues were evaluated by histopathological, immunohistochemical, and immunofluorescent examinations. Serum dihydrotestosterone and FSH levels decreased significantly in the ACR group compared to the control group, while LH levels increased and a high dose of Se prevented these changes caused by ACR. A high dose of Se prevented these changes caused by ACR. ACR-induced testicular oxidative stress, inflammation, apoptosis, changes in the expression of reproductive enzymes, some changes in sperm motility and morphology, DNA, and tissue damage, and Se administration prevented these pathologies caused by ACR. As a result of this study, it was determined that Se prevents oxidative stress, inflammation, apoptosis, autophagy, and DNA damage in testicular toxicity induced by ACR in rats.