Effect of Colchicine in reducing MMP-9, NOX2, and TGF- ß1 after myocardial infarction.

BACKGROUND: According to WHO 2020, CAD is the second leading cause of death in Indonesia with death cases reaching 259,297 or 15.33% of total deaths. Unfortunately, most of the patients of CAD in Indonesia did not match the golden period or decline to be treated with Percutaneous Coronary Intervention (PCI). Based on the recent study, there were increases in MMP-9, NOX2, and TGF-ß1 in STEMI patients which contribute to cardiac remodeling. Moreover, there is controversy regarding the benefit of late PCI (12-48 hours after onset of STEMI) in stable patients. Lately, colchicine is widely used in cardiovascular disease. This study was conducted to explore the effect of colchicine to reduce MMP- 9, NOX2, and TGF-ß1 levels after myocardial infarction in stable patients. METHOD: In this clinical trial study, we assessed 129 STEMI patients, about 102 patients who met inclusion criteria were randomized into four groups. Around 25 patients received late PCI (12-48 h after the onset of chest pain), optimal medical treatment (OMT) for STEMI, and colchicine; 24 patients received late PCI and OMT; 22 patients didn't get the revascularization (No Revas), OMT, and colchicine; and 31 patients received No Revas and OMT only. The laboratory test for MMP-9, NOX2, and TGF-ß1 were tested in Day-1 and Day-5. The data were analyzed using Mann-Whitney. RESULTS: A total of 102 patients with mean age of 56 ± 9.9, were assigned into four groups. The data analysis showed significant results within No Revas + OMT + Colchicine group versus No Revas + OMT + Placebo in MMP-9 (Day-1: p = 0.001; Day-5: p = 0.022), NOX2 (Day-1: p = 0.02; Day-5: p = 0.026), and TGF-ß1 (Day-1: p = 0.00; Day-5: p = 0.00) with the less three markers in OMT + Colchicine group than OMT + Placebo group. There were no significant differences within the late PCI + OMT + colchicine group and PCI + OMT + Placebo group. CONCLUSIONS: Colchicine could significantly reduce MMP-9, NOX2, and TGF-ß1 levels in stable STEMI patients. So that, colchicine could be a potential agent in STEMI patients and prevent cardiac remodeling events.

Colchicine as potential inhibitor targeting MMP-9, NOX2 and TGF-ß1 in myocardial infarction: a combination of docking and molecular dynamic simulation study.

The global data revealed that myocardial infarction (MI) in coronary heart disease has been the leading cause of mortality worldwide in both developing and developed countries. The remodeling process after MI is essential to be the leading cause of heart failure due to cardiac remodeling. The evidence showed the increment of MMP-9, NOX2 and TGF-ß1 expressions are biomarkers that influence cardiac remodeling. Lately, colchicine is widely used in the treatment of cardiovascular diseases. The effects of colchicine on NOX2, MMP-9 and TGF-ß1 in the molecular models are still not yet discussed. We proposed a molecular docking and molecular dynamics simulation study to show the interaction between colchicine, NOX2, MMP-9 and TGF-ß1. Colchicine has a good binding affinity with MMP-9, NOX2 and TGF-ß1 based on the value, which are -8.3 Kcal/mol, -6.7 Kcal/mol and -6.5 Kcal/mol, respectively. Colchicine also binds to some catalytic residues in MMP-9, NOX2 and TGF-ß1 that are responsible for inhibitor effects. The RMSD values between colchicine and MMP-9, NOX2 and TGF-ß1 are 2.4 Å, 2 Å and 2.1 Å, respectively. The RMSF values of ligand and receptors complex showed relatively similar fluctuations. The SASA analysis showed that colchicine could create a more stable interaction with MMP-9. PCA analysis revealed that colchicine is capable of creating a solid and stable interaction with MMP-9 mainly, also NOX2 and TGF-ß1. In conclusion, docking and molecular dynamics analysis showed evidence of colchicine roles in the inhibition of MMP-9, NOX2 and TGF-ß1 in order to inhibit the remodeling process after MI.Communicated by Ramaswamy H. Sarma.

The role of methylenetetrahydrofolate reductase C677T gene polymorphism as a risk factor for coronary artery disease: a cross-sectional study in the Sidoarjo Regional General Hospital.

Introduction: hyperhomocysteinemia (HHcy) may contribute to an increased risk of coronary artery disease (CAD). The underlying mechanisms are not well understood, but other than dietary intake factors, hyperhomocysteinemia may genetically result from a methylenetetrahydrofolate reductase (MTHFR) C677T gene polymorphism. A cross-sectional study was performed to assess whether this mutation was a potential genetic risk factor for CAD. Methods: this cross-sectional study was performed on 30 CAD patients and 30 normal healthy controls at Sidoarjo Regional General Hospital. The polymorphisms of the MTHFR C677T gene was assessed by polymerase chain reaction (PCR), and plasma homocysteine was measured by chemiluminescence immunoassay (CLIA) and then compared between CAD patients and control subjects by the multivariate logistical regression model. Results: results from an independent sample t-test analysis showed that plasma homocysteine concentrations were significantly higher in CAD patients compared to the control group individuals (13.91 ± 4.55 µmol/L vs 10.97 ± 3.45 µmol/L; p<0.05). There were no significant correlations between MTHFR C677T gene polymorphism and other risk factors, such as age at diagnosis with acute coronary syndrome, sex, smoking, lipid profile, diabetes, hypertension, C-reactive protein (CRP), creatinine, and homocysteine (p>0.05). In multivariate analysis models, the C677T genotype frequencies were insignificantly different between CAD patients and control subjects (p>0.05). Meanwhile, the results of adjusted odds ratio (aOR), 95% confidence interval (CI), and p-value for homocysteine, age, and smoking were aOR: 1.264, 95% CI : 1.042-1.535, p = 0.018; aOR: 0.916, 95% CI: 0.842-0.997, p = 0.043, and aOR: 5.428, 95% CI 1.532-19.226, p = 0.009, respectively. Homocysteine, age, and smoking were significantly different between CAD patients and control subjects (p<0.05). Conclusion: hyperhomocysteinemiais significantly correlated with an increased risk of CAD, but MTHFR C677T gene polymorphism might not contribute to increased CAD risk.

Lactobacillus plantarum FNCC 0137 fermented red Moringa oleifera exhibits protective effects in mice challenged with Salmonella typhi via TLR3/TLR4 inhibition and down-regulation of proinflammatory cytokines.

BACKGROUND: Salmonella typhi is a foodborne pathogenic bacterium that threatens health. S. typhi infection exacerbated the antibiotic resistance problem that needs alternative strategies. Moringa oleifera possesses anti-inflammatory and antimicrobial effects. However, there is a lack of information about the pharmacological value of red M. oleifera. The fermentation of red M. oleifera leaves extract (RMOL) is expected to add to its nutritional value. OBJECTIVE: The present study aimed to evaluate non-fermented RMOL (NRMOL) and fermented RMOL (FRMOL) effects on S. typhi infection in mice. MATERIALS AND METHODS: Female Balb/C mice were randomly divided into eight groups. The treatment groups were orally administered with NRMOL or FRMOL at doses 14, 42, and 84 mg/kg BW during the 28 days experimental period. Then S. typhi was introduced to mice through intraperitoneal injection except in the healthy groups. The NRMOL or FRMOL administration was continued for the next seven days. Cells that expressed CD11b+ TLR3+, CD11b+TLR4+, CD11b+IL-6+, CD11b+IL-17+, CD11b+TNF-a+, and CD4+CD25+CD62L+ were assessed by flow cytometry. RESULTS: Our result suggested that NRMOL and FRMOL extracts significantly reduced (p <0.05) the expression of CD11b+TLR3+, CD11b+TLR4+, CD11b+IL-6+, CD11b+IL-17+, and CD11b+TNF-α+ subsets. In contrast, NRMOL and FRMOL extracts significantly increased (p <0.05) the expression of CD4+CD25+CD62L+ subsets. NRMOL at dose 14 and 42 mg/kg BW was more effective compared to FRMOL in reducing the expression of CD11b+TLR3+, CD11b+TLR4+, and CD11b+TNF-α+ subsets. CONCLUSION: Our findings demonstrated that NRMOL and FRMOL extracts could be promising agents for protection against S. typhi infection via modulation of TLR3/TLR4, regulatory T cells, and proinflammatory cytokines.

Screening through Temperature and Thermal Pattern Analysis in DMBA - Induced Breast Cancer in Wistar Rats.

BACKGROUND: Based on thermal temperatures around the breast, thermography is considered a promising approache providing information about the condition of the breast without any side effects. OBJECTIVE: Using thermography, breast screening is highly dependent on the process of heat recognition. The angular effects in the process of thermal patterns recognition can increase false detection. The effect can be observed in breasts with growing mammary glands. This study aims to develop a system to identify breast conditions through analysis of temperature and thermal patterns. MATERIAL AND METHODS: In this experimental study, analysis of thermal patterns are performed using the Canny method, specifically detection of anomalies in the breast. Twenty-four Wistar female rats were used as experimental animal models with group 1 (normal), group 2 (induced with DMBA), group 3 (rats with growing mammary gland). At the end of 8 weeks, all rats were sacrificed and histopathology analysis was performed. The body temperature was measured every week using the Infrared Camera type TiS20 brand Fluke camera. RESULTS: Histopathology indicated average temperature of 36.66 °C, 37.77 °C and above 38.87 °C in normal, growing mammary glands, and cancerous breasts, respectively. These results revealed significantly higher heat in breasts with cancerous lesions. In the analysis of thermal pattern recognition for breast, no curve was formed in the normal group, while cancerous and growing mammary glands demonstrated a perfectly closed curve and an imperfect curve pattern, respectively. CONCLUSION: Breast screening through the analysis of temperature and thermal patterns can distinguish normal, cancerous and breast with growing mammary glands.

Exposure of 1800 MHz Radiofrequency with SAR 1,6 W/kg Caused a Significant Reduction in CD4+ T Cells and Release of Cytokines In-Vitro.

BACKGROUND: Although there have been many studies investigating the effects of electromagnetic fields on humans cells and tissues, the effects of radiofrequency electromagnetic fields exposure on the cells of the immune system are still controversial. OBJECTIVE: To investigate the effects of 1800 MHz RF-EMF exposure on peripheral blood mononuclear cells by measuring T helper cells count and the cytokine profile under different conditions of durations and distances. METHODS: The peripheral blood mononuclear cells (PBMCs) from healthy human subjects were exposed to 1800 MHz RF-EMF, with durations of 15, 30, 45, and 60 minutes and distances of 5 and 25 cm. The effects of RF-EMF exposure on the number of CD4+ T cells, and the expression of IL-2, IL-10, and IL-17a after 48 hours of culture were evaluated using flow cytometry. RESULTS: Our findings indicated that closer distance and longer exposure inducedlower number of CD4+ T cells. Similarly the percentagesof IL-2, IL-10 and IL-17a expressing CD4+ T cells weredecreased significantly. The number of IL-2 expressing CD4+T cells wasincreased significantly as the duration of exposure was increased, but the number was decreased after 60 minutes exposure when compared with control group with no exposure. CONCLUSION: Exposure to RF-EMF for 60 minutes at 5 cm distance causes a significant reduction in the number of CD4+ T cells, IL-2, IL-10 and IL-17a expressing T cells.

The effect of laparoscopy on mast cell degranulation and mesothelium thickness in rats.

BACKGROUND: Laparoscopy induces adhesion due to ischemia-reperfusion injury. However, the detail pathomechanism is poorly understood. This study aimed to investigate the impact of laparoscopy on mast cell and mesothelium morphological changes in the rat. METHODS: Forty-nine males of Sprague-Dawley Rattus norvegicus were divided into four groups: a) control and b) intervention groups P1, P2, and P3 that underwent 60 min laparoscopic using carbon dioxide (CO2) insufflation at 8, 10, and 12 mmHg groups, respectively. Serum hydrogen peroxide (H2O2), catalase (CAT), superoxide dismutase (SOD), malondialdehyde (MDA), and oxidative stress index (OSI) levels were determined 24 h after laparoscopy. Histopathological analyses of mast cell infiltration and degranulation and mesothelium thickness in the liver, greater omentum, mesenterium, small intestine, and peritoneum were performed 7 days after the procedure. RESULTS: H2O2, MDA, and OSI levels were significantly increased in the intervention groups compared with the control (p<0.05), while the SOD and CAT levels were decreased in the intervention groups compared with the control (p<0.05). Mast cell infiltration and degranulation were higher in the intervention groups than in control (p<0.05), while the mesothelium thickness was significantly lower in the laparoscopic groups than in control (p<0.05). Interestingly, the decrease in mesothelium thickness was strongly associated with the increase in mast cell infiltration and degranulation (p<0.01). CONCLUSIONS: Our study shows that laparoscopy in rats increases mast cell infiltration and degranulation, which also results in and correlates with a decrease in mesothelial thickness.

EMSA Eritin Drives Expansion of Regulatory T Cells and Promotes T Cells Differentiation in Irradiated Mice.

Sublethal irradiation therapy in cancer treatment causes generalized immunosuppression, which results in a range of DNA damage. We examined the significance of a polyherbal medicine called "EMSA Eritin" on immunological responses in sublethally irradiated mice focusing on the involvement of Treg, naïve T cell, and also the development and differentiation of T cells in thymus. Normal BALB/c mice were sublethally irradiated with dose of 600 rad. The irradiated mice were then orally administered by EMSA Eritin once a day at different doses: 1.04, 3.12, 9.37 mg/g body weight. The treatment was performed for 14 days. On day 15, immunological responses were observed by analyzing the status of Treg and differentiation of T cells in thymus. The administration of EMSA Eritin to irradiated mice resulted in a significant increase of pre T cells, Treg cells, and naïve T cells, which in general could maintain and normalize healthy condition in mice.

Effects of 1,25(OH)2D3 in immune response regulation of systemic lupus erithematosus (SLE) patient with hypovitamin D.

Vitamin D deficiency has been associated with pathogenesis of autoimmune diseases including SLE; however, there were still lack of data about the effects of administration of vitamin D in immune regulation in SLE patients. The aim of this study was to investigate the effects of calcitriol/1,25(OH)2D3 on dendritic cells maturation and Th17 and Treg cells activation in SLE patients with hypovitamin D. The monocytes and lymphocytes of five SLE patients with hypovitamin D were divided into 4 groups, P0 (0 nM/control), P1 (1 nM), P2 (10 nM), and P3 (100 nM) as cultured samples. Flowcytometry analysis was used to evaluate dendritic cell maturation (the percentage of CD40, CD86, and HLA-DR expression) and the amount of Th17 and Treg cells (the percentage of Th17 and Treg cells). Cytokines production of IL-12, IL-17A, and TGF-ß measured by ELISA. This study showed significant differences in CD40, CD86, HLA-DR expressions, and Th17 percentage in 10 nM of 1,25(OH)2D3 compared to that of control. For cytokines secretion, there was also significant difference between IL-12p70 and IL-17A levels in 10 nM of 1,25(OH)2D3 compared to that of control. The 1,25(OH)2D3 increased Treg cells and TGF-ß level but not significant. Our study concluded that 1,25(OH)2D3 inhibited dendritic cells maturation and Th17 cells activation in SLE patients. The 1,25(OH)2D3 increased Treg cells but not significant.