Protective Effects of Curcumin on Sperm and Stereological Parameters in Testes of Formaldehyde-Exposed NMRI Mice: An Experimental Study

Background and Aims: Formaldehyde (FA) exposure is an important cause of cellular injury and oxidative damage in testis, leading to infertility. This study aimed to assess the protective effects of curcumin on sperm and stereological parameters in testes from formaldehyde-exposed NMRI mice. Materials and Methods: At 6-8 weeks of age, 24 adult male NMRI mice weighing 30-35 g were categorized into four groups (n=6) based on the treatment they received: Group І (control) received no treatment, group ΙΙ received FA (10 mg/kg), group ΙΙΙ received FA (10 mg/kg) and curcumin (100 mg/kg), and group IV (Solvent) received dimethyl sulfoxide (0.2 ml/day). Materials were administered intraperitoneally for 35 days. After excision, epididymis tissues were placed in 1 mL aliquots of Ham’s F10 medium at 37˚C for 20 min and were then used in analyses of sperm parameters. Testes were fixed and stained with Hematoxylin & Eosin to investigate stereological indices. We also determined lipid peroxidation levels using malondialdehyde assays. Results: Mean sperm parameters (count, motility, viability, and morphology) differed significantly between groups ΙΙ and ΙΙΙ (p≤0.001). Stereological indices, including Leydig and spermatogonia cell numbers and surface-to-volume ratios of seminiferous tubules were significantly higher in group ΙΙΙ than in group ΙΙ (p≤0.001) . Finally, malondialdehyde levels in group III were significantly lower than in group II (p=0.001). Conclusions: The data showed that the curcumin, as an antioxidant, reduced FA-induced damage in sperm parameters and stereological indices in mice testes.

Nonylphenol Induces Apoptosis through ROS/JNK Signaling in a Spermatogonia Cell Line

Nonylphenol (NP) is an endocrine-disruptor chemical that negatively affects reproductive health. Testes exposure to NP results in testicular structure disruption and a reduction in testicular size and testosterone levels. However, the effects of NP on spermatogonia in testes have not been fully elucidated. In this study, the molecular mechanisms of NP in GC-1 spermatogonia (spg) cells were investigated. We found that cell viability significantly decreased and apoptosis increased in a dose-dependent manner when GC-1 spg cells were exposed to NP. Furthermore, the expression levels of the pro-apoptotic proteins increased, whereas anti-apoptosis markers decreased in NP-exposed GC-1 spg cells. We also found that NP increased reactive oxygen species (ROS) generation, suggesting that ROS-induced activation of the MAPK signaling pathway is the molecular mechanism of NP-induced apoptosis in GC-1 spg cells. Thus, NP could induce c-Jun phosphorylation; dose-dependent expression of JNK, MKK4, p53, and p38; and the subsequent inhibition of ERK1/2 and MEK1/2 phosphorylation. The genes involved in apoptosis and JNK signaling were also upregulated in GC-1 spg cells treated with NP compared to those in the controls. Our findings suggest that NP induces apoptosis through ROS/JNK signaling in GC-1 spg cells.

Knockout of the Transducin-Like Enhancer of Split 6 Gene Affects the Proliferation and Cell Cycle Process of Mouse Spermatogonia

Tle6 (Transducin-like enhancer of split 6) is a member of the Tle co-repressor superfamily, which is expressed in various tissues of invertebrates and vertebrates and participates in the developmental process. However, the current research has only found that the TLE6 mutation is related to infertility, and the key regulatory mechanism of TLE6 remains to be explored. In this study, we combined Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-Cas9 and the Tet-on system to construct mouse spermatogonia cell lines that induced TLE6 protein knockout (KO), and studied the effect of Tle6 on mouse spermatogonia proliferation and the cell cycle. The results showed that, after drug induction, the Tle6 gene in mouse spermatogonia was successfully knocked out at the genome and protein levels, and the Tle6 gene knockout efficiency was confirmed to be 87.5% with gene-cloning technology. At the same time, we also found that the mouse spermatogonia proliferated slowly after the Tle6 knockout. Using flow cytometry, we found that the cells did not undergo significant apoptosis, and the number of cells in the S phase decreased. After real-time quantity PCR (qRT-PCR) analysis, we found that the expression of cell-proliferation-related genes, CCAAT enhancer-binding protein α(C/ebp α), granulocyte-colony stimulating factor(G-csf), cyclin-dependent kinases 4(Cdk 4), Cyclin E, proliferating cell nuclear antigen(Pcna), and S-phase kinase-associated protein 2 (Skp2) was significantly reduced, which further affected cell growth. In summary, Tle6 can regulate spermatogonia cell proliferation and the cell cycle and provide a scientific basis for studying the role of TLE6 on spermatogenesis.

TINGKAT KEMATANGAN GONAD IKAN KERLING JANTAN, Tor tambroides, (Cyprinidae) YANG TERTANGKAP DI DAERAH ALIRAN SUNGAI JAMBAK KABUPATEN ACEH BARAT: PENDEKATAN HISTOLOGI

The study about gonado somatic level of fresh water is database for controlling breeding and conservation in the future. Some asessment can conducted to observing gonado somatic index is with plasma levels (hormone), gonado somatic index dan histology. Tor tambroides (local name is kerling) is one of fresh water species in Aceh. However, exploitation of fish (Tor tambroides) and degradation of river habitat is endanger for this species. This study aims to knowing profile gonado somatic of Tor tambroides especially male fish with histolgy approach. Sampling of fish was conducted during 6 months i.e. July – December. For fish catching, this research was using cast net with mesh size 1 inch and 2 inch. Explorative survey was used for sampling strategy. Histology analysis was conducted at histology laboratorium (Balai Budidaya Air Payau Ujong Batee, Aceh Province). Result show that development fish gonado of Tor tambroides stated with spermatogonia cell, spermatosit primer, spermatosit secondary dan spermatid dan this result is found in every months. Nevertheless, dominant propostion value is spematogenesis with value 55%. Base on the research is known Tor tambroides (male fish) entering spawning session on July.