Circulating miR-148a-5p and miR-21-5p as Novel Diagnostic Biomarkers in Adult Egyptian Male Patients With Metabolic Syndrome.

BACKGROUND: Metabolic syndrome (MetS) is a group of abnormalities related to metabolism that increase the risk of cardiac diseases, type 2 diabetes and mortality. MicroRNAs (miRs) act as regulators of many cellular and metabolic events, and any dysregulation of these tiny molecules can cause great disturbance in one's health. The main purpose of this study was to ascertain the diagnostic potential of miR-148a-5p and miR-21-5p in MetS. METHODS: Serum levels of miR-148a-5p and miR-21-5p were quantified in 118 male patients with MetS and 30 healthy controls by quantitative real-time polymerase chain reaction. Fasting plasma glucose, serum high-density lipoprotein cholesterol and serum triacylglycerol were measured by the colorimetric method. Blood pressure and anthropometric measurements were performed on each individual. All MetS patients had diabetes and had a large waist circumference, and were divided into 3 groups: group 1, dyslipidemic and hypertensive; group 2, normotensive; and group 3, normal lipid profile. RESULTS: miR-148a-5p expression was significantly upregulated in all MetS patients: group 1, 70.3±8.07 (p<0.0001); group 2, 75.0±9.17 (p<0.0001) and group 3, 33.7±6.89 (p<0.0001), when compared with control subjects. However, miR-21-5p expression was elevated only in the sera of group 1 (36.9±8.39, p<0.0001) and group 3 (48.9±12.0, p<0.0001), when compared with controls. CONCLUSIONS: Serum levels of miR-148a-5p and miR-21-5p were higher in MetS patients than in healthy controls; consequently, these serum miRs can serve as novel biomarkers for diagnosis and prognosis of MetS.

The (FTO) gene polymorphism is associated with metabolic syndrome risk in Egyptian females: a case- control study.

BACKGROUND: Variations within fat mass and obesity associated (FTO) gene had crosstalk with obesity risk in European and some Asian populations. This study was designed to investigate FTO rs9939609 association with metabolic syndrome (MetS) as well as biochemical parameters as plasma glucose, serum triacylglycerol (TAG), total cholesterol (TC) and transaminases enzymes in Arab female population from Egypt. METHODS: In order to achieve that, FTO gene rs9939609 (A < T) was genotyped using TaqMan SNP Genotyping Assay in a total of 197 females which were enrolled in this study. Fasting levels of serum insulin, lipid profile and plasma glucose, in addition to liver transaminases were measured. The association between the genotype distribution and MetS risk was evaluated using Chi-square and logistic regression tests in a case-control design under different genetic models. RESULTS: The association of genotype distribution with MetS was significant (χ2 = 8.6/P = 0.014) with an increased odds ratio under dominant model (OR = 1.97, P = 0.029 and 95%C.I = 1.07-3.6) and recessive model (OR = 2.95, P = 0.017 and 95%C.I = 1.22-7.22). Moreover, (AA) subjects showed significant lower HDL-C levels (P = 0.009) when compared to (TT) ones. In addition, interestingly subjects with (AA) genotype have significantly higher ALT levels (P = 0.02) that remained significant after correction of major confounders as body mass index and serum triacylglycerols but not after conservative Bonferroni adjustment. CONCLUSIONS: The present study shows for first time that FTO gene rs9939609 is genetic risk factor for metabolic syndrome in Egyptian population which may help in understanding the biology of this complex syndrome and highlighted that this association may be through HDL-C component. The association of this genetic polymorphism with ALT levels needs to be studied in other populations with larger sample size.

Genotype screening of APLN rs3115757 variant in Egyptian women population reveals an association with obesity and insulin resistance.

AIMS: Apelin is one of adipokines that plays a pivotal role in energy metabolism, insulin sensitivity and vascular integrity. A definite genetic variant of apelin gene (APLN), rs3115757, was recently introduced to potentially influence apelin expression in adipocytes. The aim of our study was to explore the sights of a potential association of this functional variant with obesity traits, insulin resistance indices as well as type 2 diabetes mellitus (T2DM) prevalence. METHODS: Genotype screening for rs3115757 variant in 151 Egyptian female samples was conducted. Fasting levels of serum insulin and lipid profile, in addition to plasma glucose were measured. Cochran-Armitage trend test was used to decide the risk allele and evaluate the association between the candidate variant and obesity using a case-control design. RESULTS: The homozygous G risk allele carriers showed higher values of body mass index (BMI) and waist circumference (P=0.001,0.02, respectively) as compared to CC or CG genotypes. As for GG genotype carriers, the risk of developing morbid obesity in lean subjects, (BMI<25), is twelve times the risk in subjects carrying other genotypes (OR=12.09, 95% CI: 1.4, 104.8, P=0.024). On the other hand, GG carriers are shown to be more resistant to insulin. Significantly after correction for BMI and age effects, GG genotype carriers showed 14% and 41% increment in insulin level and resistance (OR=1.14, 95% CI: 1.06, 1.23, P=0.001), (OR=1.42, 95% CI: 1.19, 1.70, P<0.001), respectively. CONCLUSION: These results suggest a prospective role mediated by this variant in mounting obesity disorders and as significant as insulin resistance complications.

The serine protease granzyme B as an inflammatory marker, in relation to the insulin receptor cleavage in human obesity and type 2 diabetes mellitus.

Chronic inflammation and insulin resistance form hallmarks of type 2 diabetes mellitus (T2DM). An increased circulating level of the serine protease granzyme B (GzmB) is observed during prolonged inflammation and is implicated in the pathogenesis of several chronic inflammatory diseases. Moreover, insulin receptor cleavage by unknown proteases, yielding elevated levels of insulin receptor α-subunit (IRα), was observed in T2DM and was proposed as a new mechanism of insulin resistance. Therefore, a possible association between GzmB and IRα is suggested. Accordingly, this study was set to explore whether GzmB and IRα levels are altered in T2DM patients with the impact of obesity. Furthermore, we aimed to identify if GzmB contributes towards inflammation and insulin resistance through its suggested extracellular activities. All subjects were assessed for anthropometric and metabolic parameters related to obesity and T2DM. In addition, fasting plasma insulin, GzmB, interleukin-1ß (IL-1ß), and IRα levels were estimated by enzyme linked immunosorbent assay. Levels of GzmB and IRα were found to be significantly elevated in T2DM patients compared to nondiabetic subjects. In addition, GzmB levels were positively correlated with measures of obesity and insulin resistance, IL-1ß, IRα, and other metabolic parameters. While multiple linear regression analysis revealed that both T2DM and central obesity were predicting factors for GzmB, our findings reveal a possible role of GzmB in T2DM.

Acute and chronic saturated fatty acid treatment as a key instigator of the TLR-mediated inflammatory response in human adipose tissue, in vitro.

A post-prandial increase in saturated fatty acids (SFAs) and glucose (Glc) activates an inflammatory response, which may be prolonged following restoration of physiological SFAs and Glc levels--a finding referred to as 'metabolic memory'. This study examined chronic and oscillating SFAs and Glc on the inflammatory signalling pathway in human adipose tissue (AT) and adipocytes (Ads) and determined whether Ads are subject to "metabolic memory." Abdominal (Abd) subcutaneous (Sc) explants and Ads were treated with chronic low glucose (L-Glc): 5.6 mM and high glucose (H-Glc): 17.5 mM, with low (0.2 mM) and high (2 mM) SFA for 48 h. Abd Sc explants and Ads were also exposed to the aforementioned treatment regimen for 12-h periods, with alternating rest periods of 12 h in L-Glc. Chronic treatment with L-Glc and high SFAs, H-Glc and high SFAs up-regulated key factors of the nuclear factor-κB (NFκB) pathway in Abd Sc AT and Ads (TLR4, NFκB; P<.05), whilst down-regulating MyD88. Oscillating Glc and SFA concentrations increased TLR4, NFκB, IKKß (P<.05) in explants and Ads and up-regulated MyD88 expression (P<.05). Both tumor necrosis factor α and interleukin 6 (P<.05) secretion were markedly increased in chronically treated Abd Sc explants and Ads whilst, with oscillating treatments, a sustained inflammatory effect was noted in absence of treatment. Therefore, SFAs may act as key instigators of the inflammatory response in human AT via NFκB activation, which suggests that short-term exposure of cells to uncontrolled levels of SFAs and Glc leads to a longer-term inflammatory insult within the Ad, which may have important implications for patients with obesity and Type 2 diabetes.

The interplay of YKL-40 and leptin in type 2 diabetic obese patients.

Recently, YKL-40 has been identified as a new inflammatory marker of type 2 diabetes mellitus (T2DM), while leptin is one of the most important adipose derived hormones. However, the relationship between them has not been elucidated. Therefore this study aimed to study their correlation in obese T2DM patients.

Vaspin and visfatin/Nampt are interesting interrelated adipokines playing a role in the pathogenesis of type 2 diabetes mellitus.

Recently, vaspin and visfatin/Nampt have been identified as interesting novel adipokines having insulin-sensitizing and insulin-mimetic effects, respectively. However, the relationship between them has not been elucidated; and their circulating levels in type 2 diabetes mellitus (T2DM) have not been adequately studied. Therefore, this study was designed to investigate whether their levels are altered in Egyptian T2DM patients and to study the correlation of these novel adipokines with each other and with insulin resistance, interleukin-6 (IL-6), and other biochemical parameters. The levels of vaspin, visfatin/Nampt, IL-6, insulin, and other parameters were measured in nonobese and obese T2DM patients together with matched healthy nondiabetic control subjects. Vaspin, visfatin/Nampt, and IL-6 levels were measured by enzyme-linked immunosorbent assay, whereas insulin levels were measured by chemiluminescence technique. Vaspin and visfatin/Nampt levels were found to be significantly elevated in nonobese (1.62 ± 0.22 and 25.9 ± 3.44 ng/mL, respectively) and obese T2DM patients (2.76 ± 0.38 and 45.4 ± 4.60 ng/mL, respectively) compared with control subjects (0.42 ± 0.05 and 9.37 ± 1.98 ng/mL, respectively) at P < .01. In addition, vaspin and visfatin/Nampt levels were found to be significantly positively correlated with each other and with other biochemical parameters. In conclusion, both vaspin and visfatin/Nampt might play an important role in the pathogenesis of T2DM. In addition, the 3 adipokines--vaspin, visfatin/Nampt, and IL-6--are significantly interrelated with each other. Other possible mechanisms of action for vaspin should be considered besides the inhibition of unknown substrate proteases.

Elevated endotoxin levels in non-alcoholic fatty liver disease.

BACKGROUND: Emerging data indicate that gut-derived endotoxin may contribute to low-grade systemic inflammation in insulin resistant states. This study aimed to examine the importance of serum endotoxin and inflammatory markers in non-alcoholic fatty liver disease (NAFLD) patients, with and without type 2 diabetes mellitus (T2DM), and to explore the effect of treatment with a lipase inhibitor, Orlistat, on their inflammatory status. METHODS: Fasted serum from 155 patients with biopsy proven NAFLD and 23 control subjects were analysed for endotoxin, soluble CD14 (sCD14), soluble tumour necrosis factor receptor II (sTNFRII) and various metabolic parameters. A subgroup of NAFLD patients were re-assessed 6 and 12 months after treatment with diet alone (n = 6) or diet plus Orlistat (n = 8). RESULTS: Endotoxin levels were significantly higher in patients with NAFLD compared with controls (NAFLD: 10.6(7.8, 14.8) EU/mL; controls: 3.9(3.2, 5.2) EU/mL, p < 0.001); NAFLD alone produced comparable endotoxin levels to T2DM (NAFLD: T2DM: 10.6(5.6, 14.2) EU/mL; non-diabetic: 10.6(8.5, 15.2) EU/mL), whilst a significant correlation between insulin resistance and serum endotoxin was observed (r = 0.27, p = 0.008). Both sCD14 (p < 0.01) and sTNFRII (p < 0.001) increased with severity of fibrosis. A positive correlation was also noted between sTNFRII and sCD14 in the NAFLD subjects (r = 0.29, p = 0.004).Sub-cohort treatment with Orlistat in patients with NAFLD showed significant decreases in ALT (p = 0.006), weight (p = 0.005) and endotoxin (p = 0.004) compared with the NAFLD, non-Orlistat treated control cohort at 6 and 12 months post therapy, respectively. CONCLUSIONS: Endotoxin levels were considerably increased in NAFLD patients, with marked increases noted in early stage fibrosis compared with controls. These results suggest elevated endotoxin may serve as an early indicator of potential liver damage, perhaps negating the need for invasive liver biopsy. As endotoxin may promote insulin resistance and inflammation, interventions aimed at reducing endotoxin levels in NAFLD patients may prove beneficial in reducing inflammatory burden.