Reports of Biochemistry and Molecular Biology最新文献

Background: Seminal plasma exosomes are now recognized to play a complex role in the regulation of the female reproductive system infertility. The objective of this study was to assess the effect of exosomes derived from the sperm of men with oligoasthenoteratozoospermia on endometrial implantation-related genes.

Methods: To isolate the exosomes, we employed an ultracentrifugation method on samples derived from 10 fertile men with normal sperm parameters and 10 men with oligoasthenoteratozoospermia. The size distribution and ultrastructure of the exosomes were then characterized using transmission electron microscopy and dynamic light scattering. We detected an exosome marker using western blot analysis and confirmed the cytoplasmic localization of the exosomes by incubating them with DiI dye and visualizing them using fluorescence microscopy. After 6 hours of in vitro treatment of endometrial epithelial cells with 100 µg/ml seminal exosome, the endometrial receptivity genes were examined using qRT-PCR. To perform data analysis and quantification, we utilized Image J and Prism software. P< 0.05 were considered statistically significant.

Results: After 6 hours of treatment, the mRNA levels of MUC1, LIF, G-CSF, CX3CL1, and VEGF were significantly downregulated in the endometrial epithelial cells treated with oligoasthenoteratozoospermia exosomes compared to the normal group. Although changes were observed in the mean mRNA levels of IL8 and TGF-β genes in the oligoasthenoteratozoospermia group compared to the normal group, these differences did not reach statistical significance (p > 0.05).

Conclusions: Oligoasthenoteratozoospermia exosomes have a distinct effect on endometrial receptivity compared to normal exosomes, leading to reduced expression of implantation-related genes.

Background: Epithelial-mesenchymal transition (EMT) is an important physiologic process that determines the outcome of lung tissue healing after injury. Stimuli and molecular cascades inducing EMT lead to up-regulation of the mesenchymal-specific genes in the alveolar epithelial cells and to down-regulation of the genes coding for epithelial markers. Alveolar epithelial cell lines are commonly used as in vitro models to study processes occurring in the lung tissue. The aim of this study is to quantify and compare mRNA expression levels of epithelial and mesenchymal markers in a number of lung epithelial cell lines.

Methods: Lung epithelial cell lines L2, R3/1 and RLE-6TN were cultured. Repeated mRNA isolation, reverse transcription, and quantitative PCR with primers to epithelial (E-cadherin, occludin, and ZO-2) and mesenchymal (α-SMA, collagen III, and vimentin) markers were performed.

Results: First, our study revealed a higher level of epithelial transcripts in the RLE-6TN cell line compared to L2 and R3/1 cells. Secondly, we have found simultaneous mRNA expression of both epithelial (E-cadherin, occludin and ZO-2) and mesenchymal (α-SMA, collagen III and vimentin) markers in all cell lines studied.

Conclusions: Our data indicate that at the transcriptional level the L2, R3/1, and RLE-6TN cell lines are at one of the intermediate stages of EMT, which opens new possibilities for the study of EMT on cell lines. Determination of the direction of changes in epithelial and mesenchymal markers will make it possible to establish the factors responsible for both EMT and reverse mesenchymal-epithelial transition.

Background: The influence of cytokine in the reproductive system is becoming increasingly important. The polymorphisms of the transforming growth factor-β1 (TGF-β1) gene are involved in male infertility. This study aimed to demonstrate the association between TGF-β1 and infertility and to investigate its impact on semen quality.

Methods: In this case-control study, serum TGF-β1 concentration was measured in 144 patients diagnosed with infertility and 40 fertile males by enzyme-linked immunosorbent assay (ELISA). The tetra-amplification refractory mutation system-PCR (T-ARMS-PCR) analysis was performed to detect the genotyping of the TGF-β1 (+869 C/T) (rs1800470) SNPs gene.

Results: Serum concentration of TGF-β1 was less in infertile males compared to fertile ones. The detected and more effective genotypes and alleles of TGF-β1 gene polymorphic on male infertility were, in normozoospermic group, CT genotype, probability (p)= 0.45, relative risk (RR)= 1.56, confidence intervals (CI): 0.58-4.22, and T allele (p= 0.46, RR= 1.32, CI: 0.65-2.69), in oligozoospermic and azoospermic groups, CC genotype (p= 0.32, RR= 1.58, CI: 0.73-3.41), (p= 0.013, RR= 3.50, CI: 1.40-8.73), and allele C (p= 0.44, RR= 1.32, CI: 0.73-2.38), (p= 0.06, RR= 2.14, CI: 1.02-4.50), respectively. The recessive model (TT+CT) showed increased risk among normozoospermic group (p=0.44, RR=1.67, CI:0.60-4.62). The serum concentration of TGF-β1 with CT and TT genotypes was less than that of CC genotype. TGF-β1 C/T genotype correlated with low sperm number, high immotile sperm, and high abnormal sperm morphology.

Conclusions: Our study revealed that the TGF-β1(rs1800470) gene polymorphisms are associated negatively with semen quality.

Background: Environmental pollution has a profound impact on both human and animal life. Khuzestan province, which has been plagued by intense dust storms and pollution for decades, is the focus of this study. The research aims to investigate the protective effects of metformin against the toxicity of particulate matter in the livers of rats.

Methods: Male Wistar rats were selected for the study and divided into six groups: a control group, Metformin-treated groups, Iraqi dust-exposed group (Iraqi-D), Local dust-exposed group (Local-D), Iraqi dust-exposed with Metformin treatment group (Iraqi-D+Metformin), and Local dust-exposed with Metformin treatment group (Local-D+Metformin). The rats were exposed to local and Iraqi dust through a nebulizer and received oral metformin for a duration of 21 days. At the end of the intervention, liver biomarkers and oxidative stress factors were evaluated enzymatically.

Results: The study revealed that rats exposed to Iraqi and local dust experienced a significant increase in liver biomarkers, including aspartate aminotransferase (AST), alanine transaminase (ALT), and alkaline phosphatase (ALK) levels, alongside a decrease in glutathione (GSH) concentrations and an increase in malondialdehyde (MDA) levels. However, treatment with metformin was effective in preventing the increase in these biomarkers, restoring GSH levels, and averting the rise in MDA levels, as compared to the control group.

Conclusions: Exposure to particulate matter from Iraq and the local region can induce alterations in biomarkers and oxidative stress levels in the rat liver, and these effects can be mitigated through metformin treatment.

Background: Vitamin D (vit D) controls inflammation and immunity. In Behçet's disease (BD), microRNA-155 is recognized as a significant immune response regulator. We aimed to investigate the role of vit D on immunomodulation and downregulation of inflammatory pathways associated with BD and detect the role of miRNA-155 in BD.

Methods: miRNA-155 expression by Real Time -Polymerase Chain Reaction (RT-PCR), and vit D, nuclear factor Kappa-light-chain-enhancer of activated B cells (NF-κB), and Tumor necrosis fact of TNF-α) expression by Enzyme Linked Immunosorbent Assay (ELISA) were assessed.

Results: BD patients had a significantly higher relative expression of microRNA-155 (P< 0.001), it was significantly related to vascular manifestations (P< 0.001). Vit D relative expression was significantly low in BD (P< 0.001). There was a significant rise in miRNA-155 in the active group compared to the inactive group (P< 0.001). A significant decrease in vit D levels (IU) was found in inactive and active individuals suffering from BD when compared to controls (P< 0.001). A significant rise was found in vit D levels in inactive BD cases (P< 0.001). A significant positive correlations were found between miRNA-155, NF-κB, TNF-α, and negative correlations with vit D relative expression in BD patients.

Conclusions: miRNA-155 relative expression is higher in BD is significantly related to vascular manifestations. It may have a relationship to disease activity. Vitamin D relative expression is significantly low in BD patients, which can significantly influence immunomodulatory BD therapy. Vitamin D deficiency linked to active BD.

Background: Breast cancer is the most common malignancy in women worldwide. The p16 protein is a cell cycle regulator and tumor suppressor implicated in several types of cancers. However, its relationship to breast cancer is still unknown. The present study aimed to assess the association of p16 protein expression with clinicopathological features in breast cancer.This study aimed to investigate the anti-cancer effects of different gum extracts on metabolic changes and their impact on gene expression in HT-29 cell.

Methods: The study enrolled 100 patients with invasive ductal carcinoma. The samples were collected before any adjuvant chemotherapy, and p16 protein expression was determined using immunohistochemistry. Clinicopathological features were obtained from the patient's medical records.

Results: Our findings demonstrated that p16 protein expression increased in estrogen receptor-positive tumor tissues (P< 0.01). However, no significant correlation was found between the p16 protein expression and the other clinicopathological features.

Conclusions: Our study demonstrated that p16 protein expression increased in ER-positive tumor tissue from patients with invasive ductal breast carcinoma. However, no correlation was found between the p16 protein expression and the other clinicopathological features.

Background: Cinnamic acid, a phenylpropanoid acid, has been investigated as a potential alternative therapy for diabetes and its complications in some studies.

Methods: In the first stage, the viability of HepG2 cells at different concentrations of glucose and CA was assessed by MTT assay. Oxidative stress markers) CAT, GPx, GSH, and MDA) were measured spectrophotometrically. After RNA extraction, the effect of different concentrations of CA on the expression of DPP4 and inflammatory factors (IL-6, NF- κB) in HepG2 cells was assessed using real-time PCR.

Results: In HepG2 cells, CA increased catalase and glutathione peroxidase activity and GSH production in a dose-dependent manner in the presence of high glucose concentrations, with the greatest effect seen at a concentration of 75 mg/ml. Also, it reduced the amount of MDA in high-glucose HepG2 cells. Furthermore, CA decreased the expression of DPP4, NF- κB, and IL-6 genes in HepG2 cells in the presence of high glucose levels.

Conclusions: The results of our study indicated that CA reduced hyperglycemia-induced complications in HepG2 cells by decreasing inflammatory gene expression, including IL-6 and NF- κB and inhibiting the expression of DPP4, and limiting oxidative stress.

Background: The current work investigated the effect of melatonin on differentiation of adipose mesenchymal stem cells (AD-MSCs) into dopamine producing cells and its effect on autophagy process and alpha-Synuclein (α-Syn) secretion.

Methods: AD-MSCs were characterized by flow cytometry and divided into 4 groups; i) control group (AD-MSCs without any treatment), ii) M+MSCs group (MSCs treated with 1 µM melatonin for 12 days), iii) DN group (MSCs cultured in neurobasal A medium and essential neuronal growth factors for 12 days) and iv) DN+M group (MSCs cultured in neurobasal A medium and 1µM melatonin for 12 days. By the end of experiments, the dopamine and α-Syn levels using ELISA, the expression of MAP-2, m-TOR and α-Syn genes at the level of mRNA and detection of autophagosomes formation using transmission electron microscope were performed.

Results: We found that the isolated cells were MSCs due to their positivity expression for CD105 and CD90 and negativity expression for CD34 and CD45. The concentration of dopamine was significantly higher and α-Syn concentration was significantly lower in DN+M group when compared to other groups (P< 0.005). Also, this group showed the highly expression for MAP-2 gene and less expression for m-TOR and α-Syn genes (P< 0.005). Moreover, there was significantly increase in autophagosomes formation in this group than another group (P< 0.005).

Conclusions: It is concluded that the melatonin promotes the differentiation of rat AD-MSCs into dopaminergic cells via induction of autophagy process and reduction of α-Syn secretion.

Background: Background: Male infertility contributes to roughly 15% of all infertility cases in couples. The most common cause of male infertility is azoospermia, which is caused by genetic mutations. The connection between various single nucleotide polymorphisms in the PRM genes and AZF region microdeletions with male infertility has not been reported.

Methods: In this case-control study, 100 infertile males (33 with azoospermia, 48 with oligozoospermia, and 19 with severe oligozoospermia) were chosen as the study subjects, and 100 fertile males were selected. Total DNA from peripheral blood was used to amplify two sequence-tagged site markers through multiplex PCR to detect AZFc partial deletions, and SNPs in PRM1 and PRM2 were determined through PCR-RFLP. Furthermore, quantitative real-time PCR was conducted to evaluate PRM1, PRM2, and DAZ1 (found in the AZFc region) expression levels in testis tissue.

Results: The frequency of the rs779337774 SNP in the PRM2 gene in the study population had no significant differences. However, a significant association was observed between the rs737008CA genotype (P= 0.013) and the C allele (P= 0.025) as a risk factor for male infant mortality. The deletion of sY254 and sY255 was discovered in azoospermia and severe oligozoospermia patients. Furthermore, all of these genes showed considerably low expression levels. However, only DAZ1 was identified with diagnostic biomarker potential (AUC=0.742).

Conclusion: When these genes expression levels are reduced, the likelihood of spermatozoa retrieval in azoospermic individuals is elevated. Furthermore, no significant association was observed between PRM2 polymorphism and azoospermia; however, the CA genotype of PRM1 polymorphism is significantly associated with azoospermia incidence.

Background: This study aimed to investigate the GPx-1 gene polymorphism (rs1050450), the level of oxidative stress and antioxidant parameters, and the lipid profile in an obese Kurdish population in Sulaimani, Iraq.

Methods: In a case-control study,134 obese subjects and 131 normal BMI healthy individuals participated. The GPx-1 gene polymorphism was assessed by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) method. The levels of biochemical and oxidative parameters were determined using photometric methods.

Results: The results showed that the fasting blood sugar (FBS), triglycerides (TG), and low-density lipoprotein cholesterol (LDL-C) levels were significantly higher in obese subjects compared to the control group. Obese individuals had significantly lower levels of high-density lipoprotein cholesterol (HDL-C) than the controls. The GPx-1 activity and total antioxidant capacity (TAC) levels were significantly elevated in the obese group compared to the control group (P=0.006, and P<0.001, respectively). No significant difference was detected in genotype and allele frequencies of GPx-1 (rs1050450) between obese and normal BMI groups. However, the presence of the GPx-1 TT genotype enhanced the risk of obesity in females by 1.93-fold (95% CI 1.04-3.58, P=0.036). In the total population, the GPx activity increased in the presence of TT compared to CC+CT and CT genotypes.

Conclusion: The study indicated that obesity is linked to significantly higher levels of FBS, TG, LDL-C, TAC, and GPx activity and lower level of HDL-C. Also, we found the GPx-1 gene polymorphism was associated with the risk of obesity in females and increased the GPx activity.