Reports of Biochemistry and Molecular Biology最新文献

Background: Polycystic ovary syndrome (PCOS) is a common endocrine disorder and a major cause of infertility in women. Although studies have reported the effects of naringenin on PCOS; the underlying molecular mechanisms remain unclear. This study aimed to investigate the effect of naringenin on the expression of kisspeptin (Kiss1) and calcitonin gene-related peptide (Cgrp) genes in a rat model of PCOS.

Methods: Twenty female rats (180-200 g) were used in this study. To PCOS induction, two mg of estradiol valerate was injected intramuscularly (IM) per rat. The control and PCOS groups received saline, while the other two groups were treated intraperitoneally with naringenin at either 20 mg/kg or 50 mg/kg, respectively. Subsequently, hypothalamic tissue was collected, and gene expression levels were analyzed using real-time PCR.

Results: The expression Kiss1 and Cgrp genes increased significantly in the PCOS group contrasted to the control (p≤ 0/05). In the groups treated with naringenin, the levels of Kiss1 and Cgrp gene expression reduced significantly compared to the PCOS group (p≤ 0/05).

Conclusions: Naringenin may ameliorate PCOS by downregulating hypothalamic Kiss1 and Cgrp gene expression in rats. These results suggest a novel mechanism of naringenin's action and highlight its potential for clinical application.

Background: Paraquat (PQ) is a commonly used herbicide known for its high toxicity. Despite its hazardous nature, there are currently no effective treatments for PQ poisoning. This study aimed to evaluate the effects of Rosmarinic acid (RA), a phenolic compound, on PQ-induced lung injury in mice.

Methods: Mice were divided into ten groups for two experimental periods, 6- and 24-day periods (five groups each). The first group received normal saline daily, as the control group. Animals in the second group received a single intraperitoneal (i.p.) dose of PQ (25 mg/kg) on day 3. Groups three and four received RA (50 and 100 mg/kg, respectively) orally for 6 or 24 days. Group five received 100 mg/kg of RA daily. Animals were sacrificed 24 h after the last treatment, and lung samples were collected to determine histopathological changes and expression of TLR9, IL-1β, and TNF-α genes using RT-PCR.

Results: Hematoxylin and eosin staining revealed a significant reduction in lung injury following RA treatment. RA notably reduced inflammatory cell infiltration and lung tissue congestion. Furthermore, inflammatory responses triggered by PQ were suppressed after RA treatment, as demonstrated by the downregulation of IL-1β, TNF-α, and TLR9 levels.

Conclusions: These findings suggest the therapeutic potential of RA for mitigating PQ-induced lung damage and inflammation.

Background: This study examined the protective effects of zinc oxide nanoparticles (ZnO-NPs) on hepatic ischemia-reperfusion injury (HIRI) and their possible underlying mechanisms.

Methods: 48 male rats were randomly divided into six ‎groups (n=8): the sham group that received intraperitoneal normal saline solution (Sham),‎ the HIRI group, the control groups pre-treated with 5 and 10 mg/kg ZnO-NPs for 3 ‎consecutive days without surgery (ZnO5) and (ZnO10), ‎the HIRI group pre-treated with 5 mg/kg ZnO-NPs‎ for 3 consecutive days before surgery (HIRI+ZnO5), ‎and the HIRI group pre-treated with 10 mg/kg ZnO-NPs‎ for 3 ‎consecutive days before surgery (HIRI+ZnO10)‎. One hour after reperfusion, serum and tissue samples were collected for biochemical, molecular and histopathological evaluation.

Results: Administration of ZnO-NPs caused significant improvement in the elevated serum concentrations of ALT, AST, TOS and MDA, improved liver histopathology, and increased TNF-α, IL-6, and NF-κB levels in liver tissue compared to HIRI group. In addition, administration of ZnO-NPs increased the expression of miR-125 in liver tissue compared than in the HIRI group.

Conclusions: The administration of ZnO-NPs improved the effect on HIRI by enhancing miR-125b expression and suppressing oxidative stress and inflammatory cytokines.

Background: Type 2 diabetes is a complex disease resulting from interactions between genetic, epigenetic, and environmental factors. Histone deacetylases (HDAC) are essential epigenetic-regulatory enzymes that affect gene expression and, through metabolic homeostasis and beta-cell function regulation, play significant roles in the development and treatment of diabetes. In this study, we specifically focused on the effect of metformin, the first-line therapy for type 2 diabetes on the expression of class I HDAC genes.

Methods: A total of 60 patients were equally allocated into two groups: those receiving metformin treatment and those without treatment. Also, 60 subjects with normal glucose tolerance were divided into two groups: non-obese (n=30) and obese individuals (n=30). All biochemical and clinical factors were estimated using standard methods, and RT-qPCR was used to quantify the expression levels of the candidate genes in peripheral blood mononuclear cells of different groups.

Results: The metformin treatment group exhibited increased expression of HDAC1, HDAC3, and HDAC8 in comparison to the non-treatment group. Furthermore, the expression levels of HDAC 1, 2, and 3 were higher in the obese group than the non-obese. Interestingly, evaluation of biochemical and clinical factors revealed significant association between the expression of class I HDAC genes and several diabetes-related risk factors.

Conclusions: The current findings suggest that HDAC1, 3, and 8 genes expression are affected by metformin, and obesity has a substantial ability to increase the risk of diabetes. However, changes in HDAC expression may represent potential biomarkers and therapeutic targets for future clinical studies in diabetes, particularly in exploring combination therapies involving histone deacetylase inhibitors and metformin.

Background: Hashimoto thyroiditis is a chronic autoimmune disorder influenced by genetic and environmental factors. DNA methylation, regulated by DNA methyltransferase 1 (DNMT1), may play a critical role in its pathogenesis. This study investigated the association between DNMT1 polymorphism, particularly rs2228611, and gene expression in Hashimoto thyroiditis patients and also compared serum levels of thyroid-stimulating hormone (TSH) and anti-thyroid peroxidase (anti-TPO) antibodies in both affected individuals and controls.

Methods: A case-control study of 100 participants (50 Hashimoto's thyroiditis patients and 50 controls) was conducted. TSH and anti-TPO levels were measured using the enzyme-linked immunosorbent assay (ELISA). DNMT1 expression was analyzed via quantitative real time-polymerase chain reaction (qRT-PCR), while DNMT1 (rs2228611 C/T) polymorphism was assessed by high-resolution melting-polymerase chain reaction (HRM-PCR).

Results: The results revealed that Hashimoto thyroiditis patients exhibited significantly elevated serum TSH and anti-TPO levels compared to healthy controls (p < 0.0001). DNMT1 gene expression was upregulated by 1.7-fold in patients relative to controls (p = 0.04), suggesting a potential role in disease pathogenesis. Genotyping of DNMT1 rs2228611 polymorphism revealed no significant differences in allelic or genotypic frequencies between groups. However, the TT genotype showed a non-significant trend toward increased disease risk (p = 0.07). The CT genotype appeared to confer a protective effect.

Conclusions: The study's findings suggest that elevated DNMT1 expression and thyroid dysfunction are characteristic of Hashimoto thyroiditis, while the DNMT1 rs2228611 polymorphism may have a limited but possible influence, warranting further study with larger cohorts.

Background: Primary hypothyroidism (HT) has been demonstrated to be associated with oxidative stress. This study was designed to assess the role of oxidative stress in the pathogenesis of primary hypothyroidism.

Methods: The study included 97 subjects, age range (of 29-62 years); 57 of them had been diagnosed with primary hypothyroidism, and 40 healthy subjects as controls in Baghdad, during Oct 2023 to 2024. The primary HT subjects were sub-classified into the newly diagnosed primary HT group (n=24) and the established primary HT (n=33) group. Investigations encompassed serum evaluation of total antioxidant capacity (TAC), total oxidant status (TOS), 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG), NADPH oxidase-4 (NOX4), and Anti-TPO utilizing enzymatic colorimetric methods and enzyme-linked immunosorbent assay (ELISA).

Results: The median and 1^st -3^rd quartile range values of serum 8-oxo-7,8-dihydro-2'-deoxyguanosine, NADPH oxidase-4, and total antioxidant capacity levels of newly diagnosed and established primary HT were significantly elevated when correlated with those of controls (for all, p < 0.0001), with non-significant differences between both groups of primary HT. The reservoir operating characteristic (ROC) and area under curve revealed that both total oxidant status and DNA damage 8-oxo-dG had high sensitivity and specificity in differentiation between hypothyroidism patients and controls at defined cutoff values.

Conclusions: TheElevated levels of serum 8-oxodG, NOX4, and TOS reflect the underlying oxidative damage associated with reduced thyroid function and may participate to the pathogenesis of primary hypothyroidism.

Background: Chronic kidney disease (CKD) is a major cause of morbidity and mortality worldwide, often progressing silently until advanced stages. This study aimed to evaluate the diagnostic potential of serum visfatin levels and Nicotinamide Phosphoribosyl transferase (NAMPT) gene expression in peripheral blood mononuclear cells (PBMCs) among CKD patients, along with their correlation with disease severity and lipid profile.

Methods: A case-control study included 30 CKD patients, divided into two subgroups: 15 end-stage renal disease (ESRD) patients undergoing hemodialysis (HD) and 15 non-dialysis patients. These patients were matched by age and body mass index (BMI) with 30 healthy subjects (HS). Serum visfatin, lipid profile, electrolytes, NAMPT gene expression, and other biochemical markers were measured.

Results: This study showed significantly higher visfatin levels in CKD patients compared to HS, with the highest levels observed in the ESRD group undergoing HD (5.6±1.63 ng/mL compared with 3.5±1.4 ng/mL in CKD without HD, and 2.7±1.1 ng/mL in HS; p≤0.001). Similarly, NAMPT gene expression was significantly upregulated in CKD patients, with the highest expression in the HD group, correlating strongly with serum visfatin levels (r = 0.76, p≤0.001) and lipid profile markers, including triglycerides (r = 0.67, p=0.002) and low-density lipoprotein (LDL; r = 0.61, p=0.004). In CKD patients undergoing HD, visfatin levels showed a positive correlation with triglycerides and LDL levels, suggesting a link with dyslipidemia. No significant correlation was found between visfatin and highly sensitive C-reactive protein (hsCRP), urea, creatinine, or very-low-density lipoprotein (VLDL).

Conclusions: These findings indicate that serum visfatin and NAMPT gene expression could serve as novel biomarkers for assessing CKD severity, particularly in patients undergoing hemodialysis, with potential implications for managing inflammation and cardiovascular risk in CKD.

Background: The anti-oxidative and anti-inflammatory effect of memantine (an N-methyl-D-aspartate receptor inhibitor) has been shown. Therefore, the present study aimed to evaluate the preventive effects of memantine against lipopolysaccharide (LPS)-induced sub-acute lung injury in mice.

Methods: Male C57BL/6 mice (n=30) were randomized in five groups as follows: (1) control (saline containing 10% DMSO); (2) LPS (5 mg/kg, intraperitoneally); and (3, 4, and 5) LPS 5 mg/kg + memantine 5, 10, 20 mg/kg, respectively. Memantine (dissolved in 10% DMSO) was administrated orally three days before the LPS injection and continued for three days after injury induction. Finally, the levels of markers of oxidative stress, malondialdehyde (MDA), catalase (CAT) and superoxide dismutase (SOD), interleukin (IL)-1β, tumor necrosis factor-α (TNF-α), and nitric oxide (NO), were measured and histopathological changes in the lung tissue were assessed.

Results: Lipopolysaccharide (LPS) administration increased the TNF-α, IL-1β, NO, and MDA, levels, while decreasing the lung tissues activity of CAT (P< 0.05) and SOD (P< 0.001) and caused lung pathological damages. Memantine 20 mg/kg, alleviated LPS-induced injury score, reduced the lung tissue levels of TNF-α, IL-1β, MDA, and NO, and restored CAT activity (P< 0.05, P< 0.01).

Conclusions: LPS-triggered elevation of lung injury markers including histopathological changes, inflammatory cytokines, and oxidative damage. All pathological changes were suppressed by memantine.

Background: Neonatal hyperbilirubinemia affects up to 80% of infants, and despite phototherapy as standard care, zinc inhibits heme oxygenase, lowers bilirubin and CO, and offers antioxidant benefits. This study evaluated the effect of zinc supplementation on the incidence and severity of neonatal hyperbilirubinemia.

Methods: A double-blind, block-randomized trial was conducted from December 2024 to February 2025 at Prof. Dr. R. D. Kandou Hospital NICU. Neonates with serum bilirubin >5 mg/dL were randomized to receive zinc 5 mg, zinc 10 mg, or placebo, all with phototherapy. Bilirubin levels were measured at baseline, 72 hours, and 120 hours, and outcomes were analyzed using bivariate and multivariate methods.

Results: Ninety neonates, median age 3.0 days; 67% male; 62% birth weight ≥2.5 kg, were evenly distributed among groups with no significant differences at baseline. By day 3, the 10 mg zinc group showed significantly lower median bilirubin levels (5.3 mg/dL) compared to other groups (p=0.027). By day 5, bilirubin levels further declined across all groups: 3.5 mg/dL (placebo), 2.5 mg/dL (5 mg), and 2.4 mg/dL (10 mg) (p=0.025). Hyperbilirubinemia resolution by day 5 was achieved in 67% (placebo), 90% (zinc 5 mg), and 87% (zinc 10 mg) (p=0.044). Multivariate analysis revealed that 5 mg zinc significantly increased the odds of bilirubin resolution (OR 8.0; 95% CI 1.48-44.22; p=0.016), whereas 10 mg did not. Vomiting occurred in 13.3% of neonates receiving 10 mg zinc.

Conclusions: Zinc 5 mg supplementation significantly accelerates bilirubin reduction compared to 10 mg zinc or placebo in neonates with hyperbilirubinemia.

Background: Human second-degree burns can form blisters that allow burn wound microenvironment (WME) fluid to accumulate, which leads to inflammation. Different types of cells are present in burn WME, including macrophages; these innate immune cells are present in tumor microenvironments (TMEs) and burn WMEs. They adapt their phenotypes according to environmental stimuli, which vary from pro-inflammatory (M1) to anti-inflammatory (M2). It is evident that these microenvironments share some similarities in terms of macrophage plasticity; therefore, this study examines whether burn blister exudate (BBE) can enhance macrophage activity against CT-26 cancer cells by macrophage polarization.

Methods: Real-time PCR and ELISA were used to examine the effects of human BBE on untreated and M2 macrophages. As part of the immune response assessment, yeast phagocytosis was conducted. The impact of BBE-induced macrophages on CT-26 cancer cell survival and migration was assessed using MTT proliferation assay and scratch wound healing assay, respectively.

Results: According to the results, tumor necrosis factor-alpha, interferon regulatory factor 5, induced nitric oxide synthase, and CD86 were upregulated as M1-related markers and cytokines, and M2-associated cytokines and markers, transforming growth factor beta, IL-10, Fizz1, Arginase-1, and CD206, were downregulated in untreated and M2 macrophages treated with BBE. BBE also enhanced the phagocytic capacity of untreated and M2 macrophages. Furthermore, the incubated CT-26 cell line with conditioned medium of BBE treatment groups suppresses proliferation and impedes migration of cancer cells.

Conclusions: we found that BBE-treated macrophages possess an M1-like phenotype and inhibit the proliferation and motility of CT-26 cancer cells.