Background: Effective culture systems for attachment, migration, proliferation, and differentiation of spermatogonial stem cells (SSCs) can be a promising therapeutic modality for preserving male fertility. Decellularized extracellular matrix (ECM) from native testis tissue creates a local microenvironment for testicular cell culture. Furthermore, platelet-rich plasma (PRP) contains various growth factors for the proliferation and differentiation of SSCs. Methods:In this study, human testicular cells were isolated and cultured for 4 weeks, and SSCs were characterized using immunocytochemistry (ICC) and flow cytometry. Human testicular tissue was decellularized (0.3% SDS, 1% Triton), and the efficiency of the decellularization process was confirmed by histological staining and DNA content analysis. SSCs were cultured on the human decellularized testicular matrix (DTM) for 4 weeks. The viability and the expression of differentiation genes were evaluated by MTT and real-time polymerase chain reaction (PCR), respectively.Results: Histological evaluation and DNA content analysis showed that the components of ECM were preserved during decellularization. Our results showed that after 4 weeks of culture, the expression levels of BAX, BCL-2, PLZF, and SCP3 were unchanged, while the expression of PRM2 significantly increased in the cells cultured on DTM supplemented with PRP (ECM-PRP). In addition, the expression of GFRA1 was significantly decreased in the ECM group compared to the control and PRP groups. Furthermore, the MTT test indicated that viability was significantly enhanced in cells plated on DTM supplemented with PRP. Conclusion:Our study demonstrated that DTM supplemented with PRP can provide an effective culture system for the differentiation and viability of SSCs.
Objectives: This study aimed to investigate the effect of cadmium (Cd) on the matrix metalloproteinases (MMPs) -2 and -9 expression in the lung, and the role of N-acetylcysteine (NAC) in preserving the lung cells against Cd toxicity. Methods: The rats were randomly divided into five groups of G1 (control), G2 (single dose of Cd), G3 (continuous dose of Cd), G4 (single dose of Cd+NAC), and G5 (continuous dose of Cd+NAC). The level of Cd in the blood and lung tissue was measured by atomic absorption spectroscopy. Moreover, the expression of MMP2 and MMP9 genes was considered using RT-PCR. Results: Single and continuous exposure to Cd caused a significant increase in serum and the lung tissue of Cd in G2 (0.23±0.04 mg/L and 0.35±0.047 μg/g tissue) and G3 (0.50±0.068 mg/L and 0.81±0.063 μg/g tissue) groups, compared to other groups (P<0.001). The NAC supplementation significantly decreased Cd levels in the serum and lung tissue samples of rats exposed to single or continuous Cd (P<0.001). Furthermore, exposure to a single and continuous dose of Cd caused a significant increase in the MMP2 expression by 3.24-fold (P=0.003) and 11.9-fold (P<0.001), respectively. Additionally, single and continuous dose treatment of Cd led to a significant increase in the MMP9 expression by 3.20-fold (P=0.004) and 7.54-fold (P<0.001), respectively. The NAC treatments decreased the expression of MMP2 and MMP9 in the lung of rats exposed to single or continuous Cd. Conclusion: The Cd exposure was strongly associated with the accumulation of Cd and overexpression of MMP2 and MMP9 in the lung tissue. Moreover, the NAC can protect the lungs against Cd toxicity by decreasing Cd and down-regulating MMPs.
Background and Aim: Cadmium has similar chemical and physical properties to metals such as zinc and copper, can be transferred to cells via an ionic and molecular process, and disrupts cell biological functions and homeostasis. The aim of this study was to investigate the effect of cadmium chloride on serum and tissue levels of zinc and copper, Also, the protective role of N-acetylcysteine (NAC) in preventing cadmium-induced toxicity and tissue inflammation in the lung tissue of rats exposed to single-dose and continuous-dose cadmium. Materials and Methods: 30 male Wistar rats were randomly divided into five groups of six in this interventional-experimental study. The control group only received regular water and food. The first group received a single dose treatment of 80 mg/kg of cadmium chloride on the first day of the study, while the second group received a continuous dose of 2.5 mg / kg of cadmium chloride every other day for 4 weeks. On the first day of the study, the third group received a single dose of 80 mg / kg cadmium chloride along with 50 mg / kg N-acetylcysteine. For four weeks, the fourth treatment group received a continuous dose of 2. 5 mg of cadmium chloride along with 50 mg of N-acetylcysteine. Cadmium chloride and N-acetylcysteine were administered to the animals via gavage. At the end of the treatment period, blood samples from the heart were taken to determine the level of the elements. After anesthesia, lung tissue was removed from the chest and part of it was used for histological work, while the other part was used to measure element levels. Results: Treatment with cadmium chloride showed a decrease in serum copper levels in the second treatment group compared to the control group (P-value <0.01). In addition, cadmium chloride treatment in the second treatment group compared to the control group caused a significant reduction in copper tissue level (P-value <0.001). Cadmium chloride treatment reduced serum zinc levels in the second treatment group compared to the control group (P-value <0.01). Tissue level of zinc in the second treatment group compared to the control showed a significant decrease (P-value <0.001). Simultaneous treatment of cadmium chloride and N-acetylcysteine in the fourth treatment group as a continuous dose compared to the second treatment group caused a significant increase in serum copper level (P-value <0.05). And copper tissue level in the fourth treatment group increased significantly compared to the second treatment group (P-value <0. 001). Also, simultaneous treatment of cadmium chloride and N-acetylcysteine in the fourth treatment group showed that serum zinc level (P-value <0.05) and tissue level of zinc (P-value <0.01) were significantly higher than the second treatment group. Conclusion: Continuous N-acetylcysteine administration reduces the toxicity of cadmium chloride in rats exposed to a continuous dose of cadmium chloride and improves serum and tissue copper and zinc levels.
Background Breast cancer is one of the most common cancers in women, and many people get it every year. The cancer stem cells are maybe crucial role to exacerbates and relapse the breast cancer. Therefore, finding biomarkers in human secretions can be an suitable solution for early detection and neo adjuvant therapy. This study aimed to investigate the molecular events related to the cancer stem cells in breast cancer, after which we nominated a suitable MicroRNAs participates in breast cancer pathogenesis. Methods In this study, we investigated the relationship between molecular pathways using a bioinformatics approach. First, we selected the appropriate RNA-Seq datasets from the GEO database. We used Enrichr, KEGG, and Shiny GO databases to evaluate the signal pathways and gene ontology after isolating the gene expression profiles. In the next step, we used the STRING database to assess the protein network, and we used the Targetscan database to nominate the MicroRNA. Results 510 high-expression genes and 460 low-expression genes were associated with breast cancer and the cancer stem cells. Highly expressed genes were involved in the cell cycle and cellular aging pathways. On the other hand, low-expression genes were involved in the RNA transports, spliceosome, and apoptosis pathways. After evaluating the ontology of genes and the relationship between proteins, high-expression SPARC, INHBA, FN1, and GBA proteins were nominated. In the next section, the MicroRNAs related to these genes were hsa miR-9.5p, hsa miR-203.3p, and hsa miR-429. Conclusion In general, we examined more closely and more the relationship between the cancer stem cells pathway and breast cancer using a regular and accurate bioinformatics framework. Finally, we nominated suitable MicroRNAs that were involved in breast cancer stem cells.
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