The Effect of Rosmarinic Acid on Cell Viability, Steatosis, Paraoxonase-1, and Paraoxonase-3 Protein Levels in Palmitate-induced Non-alcoholic Fatty Liver Disease Model in HepG2 Cells

Aim:We aimed to investigate the effect of rosmarinic acid (RA) on cell viability, steatosis, paraoxonase (PON)1, and PON3 protein levels in palmitate-induced non-alcoholic fatty liver disease (NAFLD) model in HepG2 cells.Materials and Methods:To induce an experimental steatosis model, HepG2 cells were incubated with 1 mM palmitate for 24 hours. For the treatment, non-toxic RA concentrations were added to the cell culture medium simultaneously with the palmitate. Cell viability was evaluated by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay. To evaluate steatosis, intracellular triglyceride levels were measured and the cells were examined microscopically with Oil-Red O staining. PON1 and PON3 protein levels were measured by Western blotting.Results:1 mM palmitate caused a significant decrease in cell viability and a significant increase in triglyceride levels, but it did not significantly change PON1 and PON3 protein levels. RA caused a significant increase in cell viability and a significant decrease in triglyceride levels in the palmitate-treated cells. Similar findings with the triglyceride levels of cells were shown in microscopic examination of cells that were stained with Oil-Red O. RA did not significantly change PON1 and PON3 protein levels in neither non-treated cells nor treated cells with palmitate.Conclusion:Our study showed that RA increases cell viability and decreases steatosis, but it does not change PON1 and PON3 protein levels in palmitate-induced NAFLD model in HepG2 cells

Research of Urgent Biochemistry Test Ordering Habit

DergiPark: 481892tmsjAims: This study aims to reveal the inappropriate use of biochemical laboratory testing at Trakya University Hospital Biochemistry Laboratory, increase the awareness of the physicians and prevent time loss.Methods: This study was descriptive, retrospective and carried out by scanning data resources. Two 48-hour intervals were chosen to evaluate the test ordering habits of the physicians working at Trakya University Hospital. Between the dates of 3rd - 5th of November 2017, Trakya University Hospital Biochemistry Laboratory was working fully functionally. However, between the dates of 10th - 12th of November 2017, the automation system of the hospital was scheduled to be shutdown due to a technical error. All the physicians working at Trakya University Hospital were informed about the technical error of the automation system and were told that they would need to order only stat tests by using old-fashioned test request forms. The data of ordered tests in these two-time intervals were analyzed and compared by using frequencies and percentages as descriptive statistics. Results: The mean number of tests per patient was 23 between 3rd - 5th of November 2017 and 15.5 between 10th - 12th of November 2017. The number of patients who had at least one test order decreased only 13.1% between 10th - 12th of November 2017. The total number of departments who made at least one test order increased by one between 10th - 12th of November 2017. Conclusion: This study indicates that physicians should be more careful while ordering tests which are necessary. Therefore, there is a need for better communication between the laboratory staff and physicians that also plays a significant role in providing better health care for the patients

Caffeine Increases Apolipoprotein A-1 and Paraoxonase-1 but not Paraoxonase-3 Protein Levels in Human-Derived Liver (HepG2) Cells

Background: Apolipoprotein A-1, paraoxonase-1 and paraoxonase-3 are antioxidant and anti-atherosclerotic structural high-density lipoprotein proteins that are mainly synthesized by the liver. No study has ever been performed to specifically examine the effects of caffeine on paraoxonase enzymes and on liver apolipoprotein A-1 protein levels. Aims: To investigate the dose-dependent effects of caffeine on liver apolipoprotein A-1, paraoxonase-1 and paraoxonase-3 protein levels. Study Design: In vitro experimental study. Methods: HepG2 cells were incubated with 0 (control), 10, 50 and 200 ?M of caffeine for 24 hours. Cell viability was evaluated by 3-(4,5-Dimethyl-2-thiazolyl)-2,5- diphenyl-2H-tetrazolium bromide assay. ApolipoproteinA-1, paraoxonase-1 and paraoxonase-3 protein levels were measured by western blotting. Results: We observed a significant increase on apolipoprotein A-1 and paraoxonase-1 protein levels in the cells incubated with 50 µM of caffeine and a significant increase on paraoxonase-1 protein level in the cells incubated with 200 µM of caffeine. Conclusion: Our study showed that caffeine does not change paraoxonase-3 protein level, but the higher doses used in our study do cause an increase in both apolipoprotein A-1 and paraoxonase-1 protein levels in liver cell

The effects of melatonin on paraoxonase activity in isoproterenol-induced myocardial infarcted rats

Tıpta Uzmanlık TeziBu çalışmanın amacı, aterosklerozdan bağımsız bir deneysel miyokart infarktüs modelinde, serum paraoksonaz aktivitesini ve melatoninin bu aktivite üzerine etkisini incelemektir. Wistar albino erkek sıçanlar, kontrol, melatonin, isoproterenol ve isoproterenol+melatonin olmak üzere gruplara ayrıldı. Melatonin (10 mg/kg/gün), %4 etanol içinde çözülerek, melatonin ve isoproterenol+melatonin gruplarına yedi gün boyunca intraperitoneal olarak verildi. Miyokart infarktüsü oluşturmak için, isoproterenol ve isoproterenol+melatonin gruplarına altıncı ve yedinci günlerde isoproterenol (150 mg/kg/gün) intraperitoneal olarak verildi. Kalbin patolojik olarak incelenmesi için her gruptan birer sıçan rastgele seçildi ve onbeşinci güne kadar yaşatıldı. Diğer sıçanlardan, son isoproterenol enjeksiyondan 24 saat sonra intrakardiyak kan alındı. Serumda paraoksonaz aktivitesi, troponin I, total oksidan durum, total antioksidan durum, malondialdehit, trigliserit, total ve yüksek dansiteli lipoprotein kolesterol ölçüldü. Paraoksonaz/yüksek dansiteli lipoprotein oranı, oksidatif stres indeksi, çok düşük dansiteli lipoprotein ve düşük dansiteli lipoprotein kolesterol ve ateroskleroz indeksi formüllerden hesaplandı. İsoproterenol grubunun paraoksonaz aktivitesi, paraoksonaz/yüksek dansiteli lipoprotein oranı ve total antioksidan durum düzeyi kontrol grubundan düşük, malondialdehit, total oksidan durum ve oksidatif stres indeksi ise yüksekti (tümü için p=0.000). İsoproterenol+melatonin grubunun paraoksonaz aktivitesi ve paraoksonaz/yüksek dansiteli lipoprotein oranı isoproterenol grubundan yüksek, malondialdehit, total oksidan durum ve oksidatif stres indeksi ise düşüktü (tümü için p=0.001). Sonuç olarak, çalışmamız deneysel miyokart infarktüsü sonrası serum paraoksonaz aktivitesinin azaldığını, melatonin verilişinin ise bu azalmayı önlediğini gösterdi.AbstractThe aim of this study is to investigate serum paraoxonase activity and the effect of melatonin on this activity in isoproterenol-induced myocardial infarction model which is independent from atherosclerosis. Wistar albino male rats were divided randomly into following groups: control, melatonin, isoproterenol and isoproterenol+melatonin. Melatonin (10 mg/kg/day) solved in %4 ethanol was given intraperitoneally to melatonin and isoproterenol+melatonin groups for seven days. On the sixth and seventh days, isoproterenol (150 mg/kg/day) was given intraperitoneally to isoproterenol and isoproterenol+melatonin groups to induce myocardial infarction. One rat from each groups was randomly selected and lived to the fifteenth day for the pathologic examination of heart. Intracardiac blood was taken from other rats 24 hours after the last isoproterenol injection. Paraoxonase activity, troponin I, total oxidant status, total antioxidant status, malondialdehyde, triglyceride, total and high density lipoprotein cholesterol were measured in serum. Paraoxonase/high density lipoprotein ratio, oxidative stress index, very low density lipoprotein and low density lipoprotein cholesterol and atherogenic index were calculated from the formulas. Isoproterenol group?s paraoxonase activity, paraoxonase/high density lipoprotein ratio and total antioxidant status level were lower and malondialdehyde, total oxidant status and oxidative stress index were higher than control group (p=0.000 for all). Isoproterenol+melatonin group?s paraoxonase activity and paraoxonase/high density lipoprotein ratio were higher and malondialdehyde, total oxidant status and oxidative stress index were lower than isoproterenol group (p=0.001 for all). As a result, our study indicated that serum paraoxonase activity decreases after miyocardial infarction and melatonin administration prevents this decrease

Association of MG53 with presence of type 2 diabetes mellitus, glycemic control, and diabetic complications.

ObjectivesMitsugumin 53 (MG53) is a myokine that acts as a membrane repair protein in tissues. Data on the effect of MG53 on insulin signaling and type 2 diabetes mellitus (T2 DM) are still unknown; most are from preclinical studies. Nevertheless, some researchers have argued that it may be a new pathogenic factor, and therapies targeting MG53 may be a new avenue for T2 DM. Our study aims to evaluate the relationship of circulating MG53 levels with the presence of diabetes, diabetic complications, and glycemic control.MethodsWe conducted a case-control study with 107 patients with T2 DM and 105 subjects without insulin resistance-related disease. Concurrent blood samples were used for serum MG53 levels and other biochemical laboratory data. MG53 concentration was measured using Human-MG53, an enzyme-linked immunosorbent assay kit (Cat# CSB-EL024511HU).ResultsWe found no difference in MG53 levels between the diabetic and control groups (p = 0.914). Furthermore, when the subjects were divided into tertiles according to their MG53 levels, we did not find any difference between the groups in terms of the presence of diabetes (p = 0.981). Additionally, no correlation was observed between weight, BMI, waist circumference, systolic and diastolic blood pressure, fasting blood glucose, HbA1c, albumin excretion in the urine, e-GFR levels, and MG53. Finally, MG53 levels were similar between the groups with and without microvascular and macrovascular complications of diabetes.ConclusionOur research finding provides insightful clinical evidence of lack of association between the levels of MG53 and T2 DM or glycemic control, at least in the studied population of Turkeys ethnicity. However, further clinical studies are warranted to establish solid evidence of the link between MG53, insulin resistance and glycemic control in a wider population elsewhere in the world