Cellular and Molecular Biology

Bone morphogenetic protein signaling pathway: an essential role in intestinal homeostasis and diseases

2025-10-06T10:25:01+02:00

The bone morphogenetic protein (BMP) signaling pathway is a fundamental regulator of intestinal homeostasis, orchestrating the delicate balance between stem cell proliferation and epithelial differentiation along the crypt-villus axis. In opposition to the Wnt signaling pathway, BMP signaling promotes epithelial maturation and inhibits excessive stem cell expansion, thereby ensuring proper renewal and functional integrity of the intestinal epithelium. Both epithelial and mesenchymal cell populations actively contribute to BMP signaling; mesenchymal cells serve as a primary source of BMP ligands and antagonists, while epithelial cells predominantly express BMP receptors and downstream effectors. This dynamic epithelial-mesenchymal dialogue establishes and maintains the intestinal stem cell niche and structural organization of the crypts. Dysregulation of BMP signaling has been increasingly implicated in the pathogenesis of inflammatory bowel diseases (IBD) such as Crohn’s disease and ulcerative colitis, as well as in colorectal cancer (CRC). In inflammatory conditions, modulation of BMP ligands and antagonists influences epithelial regeneration and immune responses, highlighting their potential anti-inflammatory and anti-fibrotic roles. Conversely, in colorectal carcinogenesis, alterations in BMP pathway components—including mutations in BMP receptors and Smad effectors, alongside aberrant expression of BMP antagonists like Gremlin-1—disrupt the balance of intestinal homeostasis, promoting tumor initiation, progression, and metastatic potential via both canonical and non-canonical signaling mechanisms. This review comprehensively summarizes the current understanding of BMP signaling in intestinal physiology and pathology, emphasizing the critical interplay between epithelial and mesenchymal compartments, and the impact of the genetic context and pathway modulators. Further elucidation of BMP pathway dynamics promises novel therapeutic strategies for intestinal diseases through targeted modulation of this pivotal signaling cascade.

TNF-α 308 (rs1800629) and INF-γ +874 polymorphisms in dengue progression: genotype-specific trends amidst allelic non-association in West Africa

2025-10-06T10:23:56+02:00

Dengue, transmitted by Aedes mosquitoes, represents a significant global health challenge due to its complex host-pathogen interactions and varying disease severity. Genetic factors are known to influence the clinical outcome of dengue infections. This study aimed to investigate the potential role of TNF-α gene polymorphism 308 (rs1800629) and INF-γ +874 (rs62559044) in the progression of dengue virus infection. Conducted in the Central region of Burkina Faso, this study included 246 participants, comprising 117 controls and 129 dengue-positive patients. Genotyping of the TNF-α 308 (rs1800629) and INF-γ +874 A/T (rs62559044) polymorphisms was performed using restriction fragment length polymorphism (RFLP) and Amplification Refractory Mutation System PCR (ARMS-PCR) techniques, respectively. Our analysis revealed no significant correlation between lymphocyte count and dengue severity (P = 0.95). Although we did not find an association between the alleles of the SNPs TNF-α 308 (rs1800629) and INF-γ +874 (rs6255904) studied with either DF or severe DS, we cannot conclude the same for their respective genotypes Thus, the AA and GG genotypes of TNF-α are associated with the contraction of DF and DS, respectively; the former is even associated with the progression of DF to the severe form of the disease. For INF-γ AA genotypes are more associated with progression to severe dengue and the AT heterozygote could be associated with a possibility of preventing progression to DS forms. . The A allele frequencies was higher frequency in DF than in DS pour TNF-α 308 , but this difference lacked statistical significance (P > 0.005). With INF-γ tTT genotype was more prevalent in DS, whereas the AT genotype frequencies differed between DF (23.96%) and DS (19.35%). Our results reveal through the allelic levels of TNF-α 308 and INF-γ +874; that the latter would not play a significant role in the progression of dengue virus infection to severe forms. However, previous studies through a clear mechanism show a strong association between the concentrations of these cytokines and the pathogenesis of dengue. This underlines the need for further investigations to elucidate the genetic determinants of the severity of dengue. In particular, a proteomic study coupled with sequencing on a representative population of the West African region would be a great asset in understanding the involvement of the genes of these cytokines in the pathogenesis of dengue.

LncRNA MEG3 overexpression modulates proliferation without inducing apoptosis in rat cardiomyoblast h9c2 cells: a transcriptomic approach

2025-10-06T10:24:06+02:00

Long non-coding RNAs (lncRNAs) have been implicated in various biological processes including cell proliferation and apoptosis. However, the role of lncRNA-MEG3 in rat cardiomyoblast H9C2 cells remains unclear. In this study, H9C2 cells were genetically modified to overexpress lncRNA MEG3. The proliferation of these cells was evaluated using the Cell Counting Kit-8 (CCK-8) assay and direct imaging techniques. Apoptosis was assessed through flow cytometry, employing Annexin V and propidium iodide (PI) staining. Quantitative PCR was utilized to confirm the overexpression of lncRNA MEG3. Further, differential expression and alternative splicing analyses were conducted using comprehensive transcriptome sequencing. The overexpression of lncRNA MEG3 in H9C2 cells led to a significant reduction in cell proliferation, as evidenced by lower absorbance readings in the CCK-8 assay and reduced cell confluency in imaging analyses. However, flow cytometric analysis revealed no substantial differences in apoptosis between the lncRNA MEG3 overexpressing group and the control. Transcriptomic analyses demonstrated significant changes in gene expression and alternative splicing patterns, highlighting the intricate role of lncRNA MEG3 in cellular regulatory mechanisms. In conclusion, lncRNA MEG3 overexpression in rat cardiomyoblast H9C2 cells significantly inhibits cellular proliferation without markedly inducing apoptosis, suggesting a specific regulatory role in cellular growth processes. The transcriptomic alterations observed underscore the potential of lncRNA MEG3 as a key player in the molecular dynamics of cardiomyoblasts.

Chitinase from Bacillus sp. SRTI8: production, purification and biocontrol activities

2025-10-06T10:24:48+02:00

From sand in the Algerian Sahara, an isolated strain of Bacillus called Bacillus sp. SRIT8 showed little chitinase activity when grown in a minimal medium supplemented with chitin (2.36 U). Using Plackett-Burman and Box-Behnken statistical plans, we could maximize chitinase synthesis, which led to a notable increase in this enzymatic activity (112 U). The purification of the resulting enzyme involved three steps: ammonium sulfate precipitation, molecular exclusion chromatography, and anion exchange chromatography. This process yielded a specific activity of 5437.14 U/mg with a purification yield of 22.44%. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis examination revealed a protein band of about 31 kDa, and optimum enzyme activity was found at pH 5 and 40 °C. Enzyme activity was boosted by Ca⁺², Na⁺, and Mn⁺² ions but was suppressed by Hg⁺² ions. The purified enzyme inhibited the growth of the plant pathogen Fusarium graminearum on wheat in both in vitro tests. So, it might prevent fungal infections in wheat throughout the germination process. The enzyme was also effective as a bioinsecticide, killing up to 52% of the larvae of Sitophilus granarius Linnaeus, an insect pest of stored grain. Our chitinase's capacity to hydrolyze fungus cell walls as well as insect cuticles can be utilised as biological control agent.

Reciprocal regulation of mir-10b, aurora-a, p53, and e-cadherin in cisplatin resistance and cell invasion of lung cancer

2025-10-06T10:29:42+02:00

Lung cancer remains a leading cause of cancer-related deaths in developed nations, including Taiwan, mainly
due to drug resistance or malignant conditions such as metastasis. Abnormal expression levels of oncogenes
or tumor suppressors are recognized as inducing changes and malignancy in various cancers, including lung
cancer. This report illustrates that mir-10b, Aurora-A, N-cadherin, and vimentin expression levels are elevated.
At the same time, p53 and E-cadherin are reduced in A549cisR clones compared to parental cells, indicating
a possible regulatory network among these molecules in malignant neoplasms. A functional assay demonstrated
that the reduction of mir-10b or Aurora-A lessened both the resistance to cisplatin and cell motility
of A549cisR cells, accompanied by a decrease in vimentin and N-cadherin, while an increase in p53 and
E-cadherin. The co-expression of mir-10b agomir restores the drug-resistant and invasive motility of Aurora-
A-knockdown A549cisR cells. Besides, ectopic expression of the active form of Aurora-A also increases
mir-10b, N-cadherin, and vimentin expressions while decreasing E-cadherin and p53 levels, thus restoring
cisplatin resistance and cell motility in mir-10b-knockdown A549cisR cells. Interestingly. Ectopic expression
of E-cadherin reduced both motility and resistance to chemotherapeutic drugs, accompanied by altering
Aurora-A, p53, and mir-10b levels in A549cisR cells. Subsequent investigations revealed that mir-10b secretion
increased in A549cisR cells. Parental A549 cells cultured with a conditioned medium of A549cisR cells
showed significantly reduced endogenous p53 expression, while inducing Aurora-A expression and increased
viability after cisplatin treatment. Transfection with mir-10b antagomir reversed the expressions of Aurora-
A, N-cadherin, vimentin, p53 and E-cadherin and the effects in parental A549 cells cultured in a conditioned
medium. These findings suggest that the mir-10b-Aurora-A-E-cadherin pathway is crucial in orchestrating
various malignancies, such as invasive motility and drug resistance, offering a possible malignancy-priming
route in lung tumor cells with regular cisplatin treatment.

Biogenic synthesis and characterization of MgO nanoparticles from banana peel extract with evaluation of their antibacterial and antioxidant activities in cellular models

2025-10-06T10:25:10+02:00

The current research involved the preparation of Magnesium Oxide Nanoparticles (MgO NPs) by aqueous extraction of banana peels, then characterizing the resulting particles and incorporating them into polymer matrices to provide antimicrobial activity for packaging materials. The resulting particles were characterized by ultraviolet-visible (UV-Vis) and scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-ray diffraction (XRD), and infrared (FTIR) spectroscopy. The ultraviolet spectrum of Magnesium Oxide nanoparticles showed sharp absorption peaks at 350 and 500 cm⁻¹. Both SEM and TEM revealed MgO NPs as almost spherical granular structures. XRD showed six main peaks of the crystalline mineral elements of the magnesium oxide nanoparticles. FTIR results confirmed that biologically active compounds act as reducing and capping agents for the nanoparticles. The magnesium oxide nanoparticles also showed antibacterial and antioxidant activity and non-toxicity.

Genetic polymorphisms and immune cytokine profiles in the cellular pathogenesis of polycystic ovary syndrome among Iraqi women

2025-10-06T10:25:20+02:00

Immune dysregulation and genetic polymorphisms are recognized as critical contributors to the pathogenesis of polycystic ovary syndrome (PCOS). The present study aimed to investigate the roles of specific cytokines and the CYP11A1 (rs4077582) polymorphism in Iraqi women diagnosed with PCOS. The current study included collecting samples from 100 women with PCOS and 100 healthy women as a control group. Clinical and laboratory tests were conducted at Diwaniyah General Hospital. The concentration of interferon-gamma, interleukin-2, interleukin-4 and interleukin-10 in the serum was determined utilizing the ELISA technique, while genetic CYP11A1(rs4077582) polymorphisms were determined using the PCR-RFLP technique. ELISA results demonstrated the concentration of interferon-gamma, interleukin-2 in FF in PCOS are significantly increased (123.8 ± 33.6, 45.77 ± 8.92 ng/ml respectively) compared with those in controls (23.5 ± 4.29, 7.31 ± 1.07 ng/ml respectively) while concentration of IL-4 and IL-10 reduced in patients (22.62 ± 6.24 and 5.81 ± 1.11 ng/ml respectively) compared to control (167.1 ± 18.62 and 37.54 ± 7.11ng/ml respectively). The results showed an increase in rate of mutant genotype CC and allele C in patients (34% and 64% respectively) compared to controls (12% and 25% respectively), while a decrease in the proportion of normal genotype TT and allele T in patients (6% and 36% respectively) compared with healthy subjects (62% and 75% respectively). immune function of IFN-γ, IL-2, IL-4 and IL-10 and CYP11A1(rs4077582) polymorphisms significantly associated with PCOS.

Inverse correlation of miR-196a and HOXB13 expression in Iraqi patients with prostate cancer

2025-10-06T10:25:28+02:00

Prostate cancer (PCa), a significant health concern in aging men, is influenced by the HOXB13 gene, a key regulator of prostate development and cell differentiation. Dysregulation of HOXB13, including genetic variants and altered expression, is related to PCa risk and progression. Importantly, the expression of HOXB13 is modulated by microRNAs, particularly miR-196a. The miR-196a controls expression of genes via mRNA binding, causing degradation or inhibiting translation. It targets HOX genes, crucial for development, and exhibits variable activity in cancers, including PCa. Therefore, the interplay between PC, HOXB13, and miR-196a contributes to the understanding of the molecular basis of PCa and identifies potential therapeutic strategies. This study involved a case-control design, comprising 120 blood samples divided into 60 PCa patients and 60 controls. Molecular analyses were performed on these samples, involving total RNA extraction followed by purification via a commercial kit. Subsequently, complementary DNA was synthesized. A quantitative real-time polymerase chain reaction was used to measure the expression levels of the HOXB13 gene and miR-196a. Relative expression levels of both HOXB13 and miR-196a were determined using established quantification methodologies. Statistical analyses were conducted using SPSS and GraphPad Prism software. Our findings demonstrated a significant increase in HOXB13 gene expression (p≤0.01), specifically a fourfold elevation in PCa patients compared to healthy individuals. In contrast, miR-196a expression exhibited a significant decrease (p≤0.01), suggesting a potential inverse regulatory correlation with HOXB13. This study reveals a significant inverse correlation between HOXB13 and miR-196a expression in PCa patients. Specifically, the HOXB13 expression level was upregulated, while the miR-196a level was downregulated. These findings suggest that miR-196a may be used as a prospective tumor suppressor in PCa by negatively regulating HOXB13, thereby inhibiting cell proliferation and invasion. Consequently, miR-196a may emerge as a promising diagnostic molecular target for prostate cancer.

Molecular docking study and ADMET prediction of the effects of some food alkaloids on thyroxine homeostasis through their interactions with human thyroxine-binding globulin

2025-10-06T10:28:13+02:00

Thyroid hormones (THs) play a vital role in several physiological functions of the body. At the circulatory level, thyroxine (T4) has been found to be the predominant form of THs. The distribution of T4 in the blood is mainly carried out by human thyroxine-binding globulin (hTBG). This process can be interfered with by various natural substances present in foods, particularly alkaloids. Some of these alkaloids have been shown to possess a cyclic chemical structure similar to that of T4. It has therefore been hypothesised that this class could potentially compete with T4 transport in the bloodstream. A molecular docking study and ADMET prediction were performed with ten selected food alkaloids. Predicted ADMET analysis revealed that all compounds tested had adequate solubility, high human gastrointestinal absorption and minimal risk of hepatotoxicity and cardiotoxicity. Molecular docking data showed that piperine, nigellidine, capsaicin, nigellicine, and 3-hydroxyquinine had a high affinity for hTBG, with respective binding energies of - 8.1, - 8.0, - 7.7, - 7.2 and - 7.1 kcal/mol. This finding indicates that these alkaloids were successfully positioned in the binding site of hTBG and had the ability to compete with T4 and increase its free level in the bloodstream. Therefore, two suggestions can be withdrawn, depending on the physiological state of the thyroid gland. Overconsumption of these alkaloids may lead to an imbalance in T4 homeostasis in both healthy individuals and hyperthyroid patients. Conversely, this competitive dynamic may offer a therapeutic advantage in the management of hypothyroidism.

Bitter frankincense water modulates hepatic enzyme activity in female patients with irritable bowel syndrome

2025-10-06T10:28:22+02:00

This study aimed to determine and know the effect of bitter frankincense water on the levels of the functional liver enzymes in female patients with irritable bowel syndrome. This study was conducted in the consulting laboratories for pathological analyses in Iraq for the period from January to February 2025. The study was applied to 25 female patients, whose ages ranged from 20 to 65 years and healthy group consisted of 25 females, aged between 20 – 65. Bitter frankincense water, was bought from reputable local markets, enough to feed 25 female patients for two months. The amount was 3 ml/kg, and each female patient was given the dose according to his weight. This is done by dividing the dose in half, for the morning and for the evening. Before administering bitter frankincense water to female patients with irritable bowel syndrome, a five milliliter blood sample was obtained, and the liver enzymes were evaluated for the samples, alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) and gamma-glutamyl transferase (GGT). Subsequently, samples were collected from the same group of female patients with irritable bowel syndrome to compare the effects of bitter frankincense water before and after giving. The results of the study showed a significant decrease (P<0.05) in the levels of liver function enzymes, alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) and gamma-glutamyl transferase (GGT) in patients with irritable bowel syndrome in women after these patients took doses of FWB. The study's notable reduction in liver enzyme levels suggests that FWB may be a useful natural hepatoprotective treatment for people with IBS. These results encourage more investigation into FWB as a supplemental treatment for enhancing gut-liver axis balance and liver function in IBS patients.

Identification of novel variants with predicted pathogenicity as key targets in esophageal cancer

2025-10-06T10:28:34+02:00

Esophageal cancer (EC) remains a major global health challenge due to its aggressive nature and poor prognosis. Genetic alterations play a crucial role in tumor progression; however, a deeper understanding of the genetic landscape of EC is essential for identifying novel and potent therapeutic targets. This study aims to identify key genes and their variants with potential pathogenicity driving EC progression. Whole-exome sequencing (WES) was performed on EC samples to identify missense variants. A comprehensive in-silico analysis was conducted using SIFT, FATHMM, PROVEAN, MutationTaster, and LRT to classify high-risk variants. Gene expression, mutation frequency, and prognostic relevance were analyzed using GEPIA and cBioPortal platforms. Protein stability was assessed with MuPro and I-Mutant to evaluate the impact of the identified variant, while protein-protein interaction (PPI) analysis via STRING and enrichment analysis through Metascape were performed to explore associated biological pathways. A total of 331 novel high-risk missense variants were identified across 274 genes and systematically refined, narrowing down to 23 prognostically significant variants in 11 genes (PSMC1, SCN8A, HNRNPA3, RPL23, COL5A2, TBL1XR1, TCP1, HNRNPD, CALM2, ABCC2, and HNRNPA1), which were also among the most differentially expressed in EC. Variants in these genes were predicted to destabilize their corresponding proteins, contributing to EC progression. In-silico survival analysis further indicated significantly worse outcomes for patients harboring alterations in these genes, including others. Protein stability analysis confirmed their destabilizing effects, while functional enrichment highlighted their involvement in key pathways driving tumorigenesis. This study identified 11 key DEGs harboring potentially pathogenic novel missense variants, highlighting vulnerabilities for precision-targeted therapies in EC.

Epigenetic and genetic events of oral squamous cell carcinoma: perspective on DNA methylation, silencing of tumor suppressor gene, and activating oncogenes

2025-10-06T10:28:44+02:00

Oral squamous cell carcinoma (OSCC) is a major health burden in most parts of the world, and pathogenesis of the disease is strongly associated with a complex combination between genetic mutations and epigenetic alterations. The current study will explore the DNA methylation phenomenon and how it affects the silencing of tumour suppressor genes and activate oncogenes in OSCC in a bid to explain the driving molecular process behind tumour development. We evaluated 50 samples of the OSCC tissues with 50 of the adjacent normal tissues, evaluated DNA methylation patterns with methylation-specific PCR and quantitative reverse-transcriptase PCR measured the expression of the DNA repair genes like BRCA1 and MLH1, among others. This finding showed different levels of DNA methylation of cancerous and normal tissues with hypermethylation causing the inactivation of important tumor suppressor genes and hypomethylation causing the activation of oncogenes. In addition, the downregulation of DNA repair genes was noted to be highly significant in OSCC samples indicating that genomic-instability may be related to epigenetic changes. These results demonstrate that aberrant DNA methylation is central to OSCC growth and progression, thus helping in the future use of methylation patterns that can be used as early detection, diagnosis, and prognostic biomarkers. Our findings support the idea that genetic, along with epigenetic, profiling is an issue of key importance toward comprehending OSCC biology and personalized therapeutic interventions. It is recommended that further confirmation be carried out to verify the clinical significance of such epigenetic markers in bigger cohorts, as well as testing such markers in guided therapies, which might eventually lead to better patient outcomes in the management of oral cancer.

Molecular detection of Epstein–Barr virus in invasive ductal carcinoma of the breast: a case–control study

2025-10-06T10:28:53+02:00

The uncertain contribution of Epstein–Barr virus (EBV) to the etiological processes underlying invasive ductal carcinoma (IDC) of the mammary gland, especially in relation to its molecular interactions within inflamed histological contexts, remains to be elucidated. This case–control research assessed the link between EBV infection and mammary IDC in a population of Iraqi females from Kirkuk. A total of 300 breast tissue specimens preserved in paraffin blocks were evaluated, including 150 samples diagnosed with IDC and 150 samples classified as fibroadenoma serving as controls. EBV latent membrane protein-1 expression was identified through the application of immunohistochemical staining and polymerase chain reaction methodologies. EBV positivity, defined as detection by both IHC and PCR, was observed in 7.3% of IDC cases and 4% of controls, with no statistically significant difference between groups (P=0.996). No significant association was found between EBV presence and estrogen or progesterone receptor status, while Her-2 expression differed significantly between EBV-positive and EBV-negative patients (P<0.001). EBV was more frequently detected in grade I tumors and stage II breast cancers, and older patients showed a higher prevalence of EBV infection. The results indicate that although Epstein–Barr virus (EBV) is identifiable in a fraction of invasive ductal carcinoma (IDC) breast specimens, a definitive causative relationship between EBV presence and IDC occurrence within this demographic is not established. Nonetheless, EBV detection appears to exhibit higher frequency in specific histopathological grades, clinical stages, and patient age categories.

Expression patterns and clinical significance of MMP-8, MMP-9 and MMP-13 in colorectal cancer

2025-10-06T10:29:03+02:00

Colorectal cancer (CRC) is the most common malignancy of the gastrointestinal system in the world. By identifying specific gene expression patterns that indicate CRC in the early stages, it is possible to potentially diagnose the disease in the early stages and start treatment quickly. Matrix metalloproteinases (MMPs) play a crucial role in the degradation of the extracellular matrix and tissue remodeling. Among them, MMP-8, MMP-9 and MMP-13 have been found to be upregulated in various cancers, including CRC, and are associated with tumor invasion, metastasis, and angiogenesis. This study investigated tissue expressions of MMP-8, MMP-9, and MMP-13 in CRC patients and explored their possible associations with pathological and clinical factors. 100 patients with CRC and 100 control subjects were involved in the study. Tissue and blood samples were collected. The quantitative Real-Time PCR (qRT-PCR) technique was used to assess the expression levels of the MMP-8, MMP-9, and MMP-13 in CRC tissue samples in comparison with the adjacent control tissue. Our results revealed that the expression levels of MMP-8, MMP-9, and MMP-13 were significantly up-regulated in CRC tissues compared to the adjacent control group. Analysis of patients' clinicopathological features showed a statistically significant difference in the expression levels of MMP-8, MMP-9, and MMP-13 between CRC patients with and without lymphovascular invasion (LVI) and TMN stage. ROC curve results have shown that these genes are good candidate diagnostic biomarkers in CRC. These results indicated that MMP-8, MMP-9, and MMP-13 levels may serve as potential diagnostic biomarkers for CRC.

Investigation of ovarian aging markers and hormonal regulation in menopausal transition

2025-10-06T10:29:12+02:00

Menopause and ovarian dysfunction are consequences of ovarian aging, a continuous and natural process in women. Early symptoms of ovarian aging include inadequate response to ovarian stimulation, irregular menstruation, and loss of follicular function, which collectively contribute to a decline in fertility as women age. The effects of aging on the ovaries are more pronounced than on other organs, with deterioration in the follicular pool and oocyte quality influenced by endocrine, genetic, and metabolic factors. This study aimed to examine the correlation between menopausal symptoms and serum levels of vitamin D, anti-Müllerian hormone (AMH), and other fertility-related hormones, as well as their impact on the timing of menopause. Ninety-two female participants were recruited from the Fertility Center Laboratories and AL-Saader Medical City in Al-Najaf province. A structured questionnaire assessing age, weight, height, and the severity of eleven menopausal symptoms—rated on a scale from 0 (absent) to 4 (extremely severe)—was administered. Symptoms evaluated included hot flushes, heart discomfort, sleep disturbances, muscle and joint pain, psychological issues, and urogenital problems. Serum levels of follicle-stimulating hormone (FSH), 25-hydroxyvitamin D, AMH, and luteinizing hormone (LH) were measured using the ELISA method. The results showed a significant negative correlation between serum AMH and vitamin D levels with menopausal symptoms and advancing age in postmenopausal women. Vitamin D levels were also negatively correlated with age, depression scores, and body mass index (BMI). Women with vitamin D deficiency and decreased AMH experienced more severe menopausal symptoms such as hot flushes, heart discomfort, depression, irritability, bladder problems, and musculoskeletal pain. These findings suggest that low AMH and vitamin D levels during menopause, compared to younger control women aged 20–40 years, may exacerbate menopausal symptoms. Supplementation with vitamin D, AMH, and reproductive hormone regulators, alongside lifestyle management, may benefit menopausal women suffering from fertility decline and severe menopausal symptoms.