Lifestyle Modifications and Nutraceutical Interventions in the Prevention and Management of Metabolic Syndrome

Abdominal obesity, dyslipidemia, hypertension, and hyperglycemia are metabolic risk factors that are grouped together to define metabolic syndrome (MetS). It is now widely recognized that MetS is linked to a higher risk of type 2 diabetes (T2DM) and cardiovascular disease (CVD). Overall, the pathophysiology of MetS initiated by the imbalance of nutrition intake and physical activity. It involves a complex interplay of insulin resistance (IR), inflammation, dysregulated adipocyte function, and genetic susceptibility, all of which contribute to the metabolic dysfunction. Lifestyle modifications play a crucial role in managing and preventing MetS. Key strategies include adopting a balanced diet like Mediterranean diet, Dietary Approaches to Stop Hypertension (DASH), or caloric restriction (CR), engaging in regular physical activity, and maintaining a healthy weight. Nutraceuticals, including polyphenols and CR-mimetic agents, improve insulin sensitivity, reduce inflammation, lower blood pressure and cholesterol levels, reducing oxidative stress, and promoting autophagy. In addition to lifestyle changes, drug therapy may be necessary for some individuals to manage specific risk factors, such as diuretics, angiotensin-converting enzyme (ACE) inhibitors, angiotensin II receptor blockers (ARB), calcium channel blockers, and beta blockers for hypertension; biguanides, sulfonylureas, dipeptidyl peptidase-4 (DPP-4) inhibitors, glucagon-like peptide 1(GLP-1) receptor agonists, sodium-glucose co-transporter 2 (SGLT2) inhibitors, and thiazolidinediones for hyperglycemia; and statins for dyslipidemia. Early diagnosis, including waist circumference and blood pressure measurement, serum cholesterol and glucose testing, and intervention, is essential to effectively manage MetS and prevent the progression of associated diseases. In conclusion, understanding the risk factors and associated risks of MetS, along with the implementation of lifestyle modifications such as dietary and nutraceutical interventions including polyphenols and CR-mimetic agents, is vital for reducing the burden of this syndrome. Early diagnosis and proactive management are key to improving long-term health outcomes.KEYWORDS: metabolic syndrome, abdominal obesity, insulin resistance, diet, nutraceutical

Intermittent Exposure to Hypobaric Hypoxia Increases VEGF, HIF-1α, and Nrf-2 Expressions in Brain Tissue

BACKGROUND: Hypoxia inducible factor-1α (HIF-1α), nuclear factor erythroid 2-related factor 2 (Nrf-2), and vascular endothelial growth factor (VEGF), play a crucial role as neuroprotective factors. Currently, there is a lack of studies examining the biomolecular responses of the brain to intermittent hypoxia resulting from various pressures. This study was conducted to investigate the physiological responses, histopathological features, and cellular adaptive responses in the brains of rats that were intermittently exposed to hypobaric hypoxic conditions.METHODS: Thirty male Sprague Dawley rats were divided into six groups: a control group and five treatment groups exposed to hypobaric hypoxia. The treatment groups were placed in a hypobaric chamber simulating an altitude of 3,048 meters for 1 hour/day for 1, 7, 14, 21, and 28 days. After exposure, brain tissue was collected for histopathological analysis and protein quantification of HIF-1α, Nrf-2, cytoglobin (Cygb), neuroglobin (Ngb), VEGF, malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT).RESULTS: In the brain, intermittent hypobaric hypoxia significantly increased HIF-1α expression (p=0.000) and its downstream proteins Cygb (p=0.000), and VEGF (p=0.001), with a peak at 14x IHH exposure compared to control. This was followed by a significant increase in Nrf-2 expression (p=0.000), SOD (p=0.000), Gpx (p=0.000), and CAT activity (p=0.000), indicating an adaptive antioxidant response. Conversely, MDA levels was decreased with prolonged exposure, suggesting reduced oxidative damage.CONCLUSION: IHH elevates HIF-1α, Nrf-2, and oxidative stress markers, triggering an adaptive antioxidant response in the rat’s brains.KEYWORDS: HIF-1α, intermittent hypobaric hypoxia, Nrf-2, oxidative stres

N-Acetylcysteine Prevents Sleep Deprivation-induced Memory Deficit in Juvenile Rats Through the Suppression of BDNF, Cortisol, Acetylcholine Levels, and Inflammatory Cytokines Expressions

oai:ojs.inabj.org:article/3445BACKGROUND: Sleep deprivation (SD) affects 20–30% of children and is known to impair cognitive functions, particularly memory. Despite its impact, there is currently no standardized treatment. Evidence from both adult animal and human suggests that N-acetylcysteine (NAC) possesses neuroprotective properties. This study was conducted to evaluate the effects of NAC on memory deficits induced by SD in juvenile rats.METHODS: Juvenile Sprague-Dawley rats were subjected to SD using the modified multiple platform method. NAC was administered intraperitoneally at doses of 100 mg/kgBW or 500 mg/kgBW. Y-maze and novel object recognition (NOR) tests were used for neurobehavioral assessment. Biochemical analyses were conducted to measure cortisol, brain-derived neurotrophic factor (BDNF), and acetylcholine (ACh) levels, using enzyme-linked immunosorbent assay (ELISA). Acetylcholine esterase (AChE) activity was measured by colorimetric method. Western blot analysis was performed to examine cAMP response element-binding protein (CREB) and phosphorylated CREB (p-CREB). Additionally, cytokine mRNA expressions were evaluated using polymerase chain reaction (PCR).RESULTS: Spontaneous alteration and discrimination ratio were decreased in SD group without treatment compared to the normal group (p<0.05). Similarly, BDNF was also decreased compared with normal group (p<0.05). Cortisol level and mRNA expression of TNF-α were increased significantly compared with normal group (p<0.05) along with a slight increase of ACh activity. Interestingly, NAC treatment mainly at the dose of 500 mg/kgBW prevented those pathological features significantly.CONCLUSION: NAC might prevent the SD-induced memory deficits by suppressing the inflammatory markers, activity of AChE, cortisol, and enhancing the level of BDNF.KEYWORDS: SD, NAC, memory, AChE, cortisol, inflammatory markers, BDN

Immunomodulatory Effect of Dioscorea esculenta L. on NF-κB, TLR-4, TNF-α, and IL-10 Expressions in LPS-stimulated RAW 264.7 Mouse Macrophages

BACKGROUND: Gene expressions of toll-like receptor 4 (TLR)-4, nuclear factor-kappaB (NF-κB), tumor necrosis factor (TNF)-α, and interleukin (IL)-10 are known to have roles in the inflammatory process and affect the regulation of the immune system. A preliminary study showed that Dioscorea esculenta L. tuber has immunomodulatory activity against macrophage phagocytosis activity and lymphocyte proliferation. However, the immunomodulatory activity of aqueous extract (AE), polysaccharide fraction (PF), and non-polysaccharide fraction (NPF) of D. esculenta L. tubers on these gene expressions have not been elucidated well. Therefore, this study was performed to determine its immunomodulatory activity by utilizing RAW 264.7 cell culture induced by lipopolysaccharide (LPS).METHODS: RAW 264.7 cells were stimulated with LPS at a concentration of 1 µg/mL for 30 minutes before incubation with non-toxic concentrations of AE, PF, NPF, positive control, and inulin at 25 and 50 µg/mL. TNF-α, IL-10, TLR-4, NF-κB, and β-actin expressions were evaluated using reverse transcription-polymerase chain reaction (RT-PCR) and were normalized with β-actin as an internal control. Triplicate experiments were performed throughout this study.RESULTS: Treatment with 25 µg/mL NPF significantly decreased the expression of NF-κB, TLR-4, and TNF-α (p<0.05). In contrast, treatment of 25 and 50 µg/mL PF significantly decreased the NF-κB expression (p<0.05). Moreover, only treatment with 50 µg/mL AE exhibited a significant increase in IL-10 expression (p<0.05).CONCLUSION: Treatment with D. esculenta L. tuber stimulated macrophage RAW 264.7 cells via NF-κB, TLR-4, TNF-α, and IL-10 expressions. NPF at 25 µg/mL has stronger immunomodulatory activity in reducing the expression of genes involved in the inflammatory process that plays a role in regulating the immune system.KEYWORDS: Dioscorea esculenta L., Immunomodulator, IL-10, NF-κB, TLR-4, TNF-α, RAW 264.7 cel

Hypomethylation of the Soluble Fms-like Tyrosine Kinase 1 (sFlt-1) Gene Promoter Region and Elevated sFlt-1 Placental Expression as Risk Factors for Preeclampsia

BACKGROUND: Preeclampsia significantly contributes to maternal and fetal morbidity and mortality worldwide, marked by an imbalance of angiogenic factors, particularly increased soluble Fms-like tyrosine kinase-1 (sFlt-1), leading to endothelial dysfunction. Epigenetic regulation, including DNA methylation of the sFlt-1 promoter, has been suggested to influence sFlt-1 expression, but the data in Indonesian population are limited. This study was perfmed to determine whether hypomethylation of the sFlt-1 promoter and elevated placental sFlt-1 expression are associated with increased risk of preeclampsia.METHODS: A case-control study was conducted involving 30 women with preeclampsia and 30 normotensive pregnant women. Subjects were selected based on eligibility criteria that included singleton pregnancy and gestational age of ≥37 weeks. DNA methylation of the sFlt-1 promoter was assessed using methylation-specific polymerase chain reaction (PCR), and sFlt-1 expression was measured by enzyme-linked immunosorbent assay (ELISA). Statistical analyses, including Mann-Whitney U, Chi-square tests, Receiver-operating characteristic (ROC) curve analysis, and multivariate logistic regression, were performed to evaluate the relationship between methylation levels, gene expression, and preeclampsia risk.RESULTS: The preeclampsia group had significantly lower methylation levels of sFlt-1 promoter and higher placental sFlt-1 expression (both p<0.001). Hypomethylation of sFlt-1 promoter (adjusted odd ratio (AOR): 21.18; 95% CI: 2.49–179.72; p=0.005), high sFlt-1 expression (AOR: 12.55; 95% CI: 1.95–80.83; p=0.008), and obesity (AOR: 11.15; 95% CI: 2.01–61.78; p=0.006) were identified as independent risk factors for preeclampsia.CONCLUSION: Hypomethylation of sFlt-1 promoter and elevated placental sFlt-1 expression are significant independent risk factors for preeclampsia. These findings suggest that hypomethylation of sFlt-1 promoter and elevated placental sFlt-1 expression may serve as potential epigenetic biomarkers for early detection and targeted intervention in preeclampsia.KEYWORDS: preeclampsia, sFlt-1, gene expression, hypomethylation, placenta, risk facto

GSH-conjugation Reduces PGV-1 Cytotoxicity and Its Ability in Downregulating N-Myc, β-catenin, and p62 Protein in Huh-6 Cells

BACKGROUND: Pentagamuvone-1 (PGV-1), a synthetic curcumin analogue, exhibits potent anticancer activity against Hepatocellular Carcinoma (HCC) by disrupting cell cycle regulation and downregulating key oncogenes such as N-Myc. Numerous studies have examined the role of glutathione (GSH) conjugation in modulating the anticancer properties of curcumin and its analogues. In contrast, the impact of PGV-1 metabolism, particularly GSH conjugation, and its implications for anticancer efficacy have not yet been elucidated. This study was performed to prepare GSH-conjugated PGV-1 (PGV-1-(GSH)2) as the model of PGV-1 metabolite and evaluate its potential distinct cytotoxicity on Huh-6 cells.METHODS: PGV-1 was synthesized via an acid-catalyzed reaction between 4-hydroxy-3,5-dimethylbenzaldehyde and cyclopentanone while PGV-1-(GSH)2 was obtained through reflux at 70oC for 2 hours. The cytotoxic effects of PGV-1 and PGV-1-(GSH)2 on Huh-6 and JHH4, two HCC cells, were assessed using a cell counting kit-8 (CCK-8) assay, while immunoblotting was performed to evaluate their impact on N-Myc and its downstream protein such as β-catenin, and p62.RESULTS: PGV-1-(GSH)2 was prepared through GSH conjugation of PGV-1 in orange color solution, as confirmed by Electrospray Ionization Mass Spectrometry (ESI-MS), Fourier Transform Infrared Spectroscopy (FT-IR), and Nuclear Magnetic Resonance (NMR) analysis. Cytotoxicity assays revealed that PGV-1-(GSH)2 exhibited less potent anticancer activity against HCC cells than PGV-1. GSH conjugation also decreased the ability of PGV-1 in downregulating the N-Myc, β-catenin, and p62 protein level.CONCLUSION: The prepared PGV-1-(GSH)2 reduces the cytotoxicity of PGV-1 and its ability on downregulating N-Myc, β-catenin, and p62 in Huh-6 cells. These findings highlight the need for further exploration about the study of PGV-1 metabolism which could affect the anticancer efficacy against HCC.KEYWORDS: curcumin, PGV-1, GSH, HCC, N-My

FGFR2 as A Prognostic and Predictive Marker in Colorectal Adenocarcinoma Based on TILs Grade

BACKGROUND: Colorectal cancer remains a serious health problem due to its high incidence and mortality rate each year. Histopathological grades and tumor-infiltrating lymphocytes (TILs) are associated with patient’s outcome. Fibroblast growth factor receptor 2 (FGFR2) overexpression is correlated with a worse prognosis in colorectal adenocarcinoma patients. Unfortunately, there are not many studies investigating the relationship between FGFR2 with histopathological grade and TILs in Indonesia. This study was conducted to analyze the correlation between FGFR2 expression with histopathological grade and TILs grade in colorectal adenocarcinoma.METHODS: Immunohistochemistry examination using FGFR2 rabbit polyclonal antibody was performed on 94 paraffin-embedded colorectal adenocarcinoma blocks and its expression was examined using a light microscope. The relationship between FGFR2 expression with histopathological grade and TILs grade in colorectal adenocarcinoma was statistically analyzed.RESULTS: Of the 94 samples examined, low grade adenocarcinoma was more common (n=76), of which 60.5% showed high FGFR2 expression. While in high grade adenocarcinoma, 83.3% of the samples showed high FGFR2 expression. In low grade TILs (n=30), 80% showed strong FGFR2 expression. While in high grade TILs (n=17), 64.7% showed weak FGFR2 expression. Based on statistical analysis, there was a significant correlation between FGFR2 expression and TILs grade (p=0.008). However, there was no significant association with histopathological grade (p=0.127).CONCLUSION: The significant correlation between FGFR2 expression and TILs grade suggests that FGFR2 may be used as a prognostic and predictive marker in colorectal adenocarcinoma.KEYWORDS: FGFR2, Colorectal adenocarcinoma, TILs, Histopathological grad

miR-200a as Potential Early-onset Biomarker, while High Nitric Oxide as Potential Late-onset Biomarker in Preeclampsia

BACKGROUND: miR-200a is known to alter trophoblast invasion and spiral artery remodeling, leading to defective placentation that causes placental hypoxia, which is the main pathomechanism in early-onset preeclampsia (EOPE). Hypoxic placentas cause systemic endothelial dysfunction that is characterized by low production of endothelial vasodilator, mainly nitric oxide (NO). On the other hand, defective placentation does not cause late-onset preeclampsia (LOPE), making the role of miR-200a expression and NO level as predictors in LOPE questionable. Therefore, this study was conducted to compare miR-200a expressions and NO levels in EOPE and LOPE to clarify their role in pathomechanism of both types of preeclampsia.METHODS: A cross-sectional comparative study was conducted in 62 preeclamptic patients (31 EOPEs and 31 LOPEs). Subjects were classified into EOPE or LOPE groups based on whether the diagnosis of preeclampsia was made at <34 or ≥34 weeks of pregnancy. miR-200a expression was analyzed using real-time polymerase chain reaction technique, and NO level was analyzed using colorimetric assay method.RESULTS: EOPE and LOPE subjects were equivalent in terms of age and parity (p=0.709 and p=0.066), but significantly difference in gestational age (p=0.000). miR-200a were expressed in 74.2% of EOPE and 41.9% of LOPE subjects (p=0.010). Median NO levels were lower in EOPE compared to LOPE subjects (23.75 vs. 106.00 µmol/L) (p=0.027), and lower in subjects with detected miR-200a compared to subjects with undetected miR-200a (62.75 vs. 132.25 µmol/L) (p=0.032).CONCLUSION: miR-200a was more expressed in EOPE compared to LOPE subjects suggesting that it might be a significant in predicting EOPE. While NO level was significantly lower in EOPE whilst higher in LOPE subjects, hence might be potential as a marker to differentiate EOPE and LOPE.KEYWORDS: miR-200a expression, NO level, early-onset preeclampsia, late-onset preeclampsi

Insulin Resistance, Adiponectin, and Dyslipidemia as Key Determinants of Metabolic and Reproductive Dysregulation in Polycystic Ovary Syndrome

BACKGROUND: Polycystic ovary syndrome (PCOS) is a common endocrine disorder associated with insulin resistance, dyslipidemia, and metabolic dysfunction. However, the predictive value of insulin resistance (HOMA-IR), dyslipidemia, and adiponectin in identifying PCOS and stratifying metabolic risk remains unclear, particularly in underrepresented populations, such as Iraqi women. This study evaluated these parameters to improve risk stratification and early intervention strategies in resource-limited settings.METHODS: This case-control study included 100 women (50 with PCOS diagnosed using the Rotterdam criteria and 50 age-matched healthy controls). Anthropometric and clinical assessments, including BMI and hirsutism scores, were performed. Fasting blood samples were analyzed for glucose and lipid profile using colorimetric assays, insulin via electrochemiluminescence immunoassay (ECLIA), and adiponectin via enzyme-linked immunosorbent assay (ELISA), and HOMA-IR was calculated to quantify insulin resistance.RESULTS: Women with PCOS exhibited significantly higher BMI, fasting glucose, insulin, and HOMA-IR values (p<0.001 for all) than controls. Dyslipidemia was evident, characterized by elevated total cholesterol, LDL-C, and triglyceride levels, as well as lower HDL-C levels (p<0.001). Adiponectin levels were markedly reduced in the PCOS group (p<0.001) and showed strong inverse correlations with HOMA-IR (r=–0.554, p<0.001) and fasting insulin (r=–0.453, p<0.001). Logistic regression indicated that HOMA-IR was the most significant predictor of PCOS (OR=28, 95% CI 4.86–161.44, p=0.0002), whereas higher adiponectin levels offered significant protective effects (OR=0.54, 95% CI 0.36–0.82, p=0.0039).CONCLUSION: Insulin resistance, dyslipidemia, and low adiponectin levels are strongly associated with PCOS and metabolic dysfunction in Iraqi women. HOMA-IR is a key predictor, whereas adiponectin may have protective effects. These findings highlight the potential of these biomarkers in risk stratification and early intervention in resource-limited settings.KEYWORDS: PCOS, adiponectin, HOMA-IR, dyslipidemi

Alpha-Actinin-3 (ACTN3) R577X Polymorphism on Brain-derived Neurotrophic Factor (BDNF) Levels of Pre- and Post-Eccentric Exercised Male Subjects

BACKGROUND: Eccentric exercise, characterized by muscle lengthening underload, elicits physiological responses, including alterations in brain-derived neurotrophic factor (BDNF) levels, crucial for neuroplasticity and exercise adaptations. The Alpha-Actinin-3 (ACTN3) gene encodes α-actinin-3, a protein in fast-twitch muscle fibers associated with explosive performance. The R577X polymorphism in ACTN3 is associated with athletic performance, particularly in power-based activities. However, its influence on the BDNF response to eccentric exercise remains unclear. This study investigated whether the ACTN3 R577X polymorphism modulates BDNF levels post-exercise.METHODS: Male subjects aged 18-30 years old, who were not involved in structured physical activity, and abstaining from alcohol and protein supplements within specified periods, were involved in this study. Subjects’ genotypes were identified using polymerase chain reaction (PCR) and classified into different ACTN3 genotypes (RR, RX, XX). All subjects underwent an eccentric exercise protocol. BDNF levels were measured pre-exercise, post-exercise, and 72 hours post-exercise using sandwich Enzyme-Linked Immunosorbent Assay (ELISA).RESULTS: Most of subjects had RX genotype (52.2%), followed by XX (39.1%) and RR genotypes (8.7%), respectively. BDNF levels decreased significantly across all time points. The RR genotype showed a decrease from approximately 270 pg/mL to 230 pg/mL, while RX and XX genotypes showed similar patterns of reduction. No significant differences in BDNF levels were observed between genotypes at any time point.CONCLUSION: Eccentric exercise leads to a consistent decrease in BDNF levels, with no significant modulation by ACTN3 genotype. These findings suggest a uniform response to exercise-induced stress across genotypes.KEYWORDS: ACTN3, BDNF, eccentric exercises, genotype, adaptatio