Altered Biochemical and Hematological Profile of Fatty Liver Disease Patients in Western Nepal

( Mahendra Prasad Bhatt ),( Basant Kumar Tamrakar ),( Amar Nagila ) 대한간학회 2018 춘·추계 학술대회 (KASL) Vol.2018 No.1

Aims: Fatty liver disease (FLD) is one the most common liver disease in the world, however, there are limited studies on biochemical and hematological features of fatty liver disease. Different laboratory tests are extremely useful in better understanding of diseases, and thereby, confirm clinical diagnosis for the better management of disease. The aim of the present study was to demonstrate the significant changes in biochemical and hematological parameters in FLD patients of western Nepal compared with control group. Methods: About 49 patients with FLD and 27 healthy control subjects from the outpatient clinic of the Internal Medicine Department, Fishtail Hospital and Research Centre Pokhara, Nepal were enrolled in the study. Hepatosteatosis is studied by abdominal ultrasonography to confirm the diagnosis of FLD. The anthropometric parameters: height, weight, waist and hip circumferences, blood pressure were measured, and body mass index (BMI) and waist/hip ratio (WHR) were calculated. Overnight fasting blood samples were collected from the patients and controls to analyze biochemical tests: lipid profile, liver function tests, renal function tests, blood sugar, HbA1c, and hematological tests: hemoglobin, WBC and platelet counts. Results: With the increased level of serum triglyceride in the patients with FLD, total cholesterol, LDL-cholesterol had a significantly increasing trend (P<0.05); whereas HDL-cholesterol was found significantly decreased in comparison to control group. The levels of alanine aminotransferase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP) and Gamma-GT were noticed significantly difference in the cases as compared to those in the controls (P<0.05). Other biochemical and hematological parameters also noticed significant difference. Conclusions: Most of the patients with FLD were found asymptomatic. The study revealed that fatty liver patients have dyslipidemias, abnormal liver function tests, increased trend of blood sugar and HbA1c, and hematological test parameters were significantly different compared to control group.

C-peptide protects against hyperglycemic memory and vascular endothelial cell apoptosis

Bhatt, Mahendra Prasad,Lee, Yeon-Ju,Jung, Se-Hui,Kim, Yong Ho,Hwang, Jong Yun,Han, Eun-Taek,Park, Won Sun,Hong, Seok-Ho,Kim, Young-Myeong,Ha, Kwon-Soo Bioscientifica 2016 The Journal of endocrinology Vol.231 No.1

<P>C-peptide exerts protective effects against diabetic complications; however, its role in inhibiting hyperglycemic memory (HGM) has not been elucidated. We investigated the beneficial effect of C-peptide on HGM-induced vascular damage <I>in vitro</I> and <I>in vivo</I> using human umbilical vein endothelial cells and diabetic mice. HGM induced apoptosis by persistent generation of intracellular ROS and sustained formation of ONOO<SUP>−</SUP> and nitrotyrosine. These HGM-induced intracellular events were normalized by treatment with C-peptide, but not insulin, in endothelial cells. C-peptide also inhibited persistent upregulation of p53 and activation of mitochondrial adaptor p66<SUP>shc</SUP> after glucose normalization. Further, C-peptide replacement therapy prevented persistent generation of ROS and ONOO<SUP>−</SUP> in the aorta of diabetic mice whose glucose levels were normalized by the administration of insulin. C-peptide, but not insulin, also prevented HGM-induced endothelial apoptosis in the murine diabetic aorta. This study highlights a promising role for C-peptide in preventing HGM-induced intracellular events and diabetic vascular damage.</P>

C-Peptide Prevents Hyperglycemia-Induced Endothelial Apoptosis Through Inhibition of Reactive Oxygen Species–Mediated Transglutaminase 2 Activation

Bhatt, Mahendra Prasad,Lim, Young-Cheol,Hwang, JongYun,Na, SungHun,Kim, Young-Myeong,Ha, Kwon-Soo American Diabetes Association 2013 Diabetes Vol.62 No.1

<P>C-peptide is a bioactive peptide with a potentially protective role in diabetes complications; however, its molecular mechanism of protection against cardiovascular damage caused by hyperglycemia-induced apoptosis remains unclear. We investigated the protective mechanism of C-peptide against hyperglycemia-induced apoptosis using human umbilical vein endothelial cells and streptozotocin diabetic mice. High glucose (33 mmol/L) induced apoptotic cell death in endothelial cells via sequential elevation of intracellular Ca<SUP>2+</SUP> and reactive oxygen species (ROS) as well as subsequent activation of transglutaminase 2 (TG2). C-peptide (1 nmol/L) prevented endothelial cell death by inhibiting protein kinase C– and NADPH oxidase–dependent intracellular ROS generation and by abolishing high glucose–induced TG2 activation, without affecting intracellular Ca<SUP>2+</SUP> levels. Consistently, in the aorta of streptozotocin diabetic mice, hyperglycemia stimulated transamidating activity and endothelial cell apoptosis that was inhibited by C-peptide replacement therapy (35 pmol/min/kg) using osmotic pumps (control and diabetes, <I>n</I> = 8; diabetes + C-peptide, <I>n</I> = 7). In addition, C-peptide prevented hyperglycemia-induced activation of transamidation activity and apoptosis in the heart and renal cortex of streptozotocin diabetic mice. Thus, C-peptide protects endothelial cells from hyperglycemia-induced apoptotic cell death by inhibiting intracellular ROS-mediated activation of TG2. Furthermore, TG2 may be a promising avenue of therapeutic investigation to treat diabetic vasculopathies.</P>

C-peptide replacement therapy as an emerging strategy for preventing diabetic vasculopathy

Bhatt, Mahendra Prasad,Lim, Young-Cheol,Ha, Kwon-Soo Oxford University Press 2014 Cardiovascular research Vol.104 No.2

<P>Lack of C-peptide, along with insulin, is the main feature of Type 1 diabetes mellitus (DM) and is also observed in progressive β-cell loss in later stage of Type 2 DM. Therapeutic approaches to hyperglycaemic control have been ineffective in preventing diabetic vasculopathy, and alternative therapeutic strategies are necessary to target both hyperglycaemia and diabetic complications. End-stage organ failure in DM seems to develop primarily due to vascular dysfunction and damage, leading to two types of organ-specific diseases, such as micro- and macrovascular complications. Numerous studies in diabetic patients and animals demonstrate that C-peptide treatment alone or in combination with insulin has physiological functions and might be beneficial in preventing diabetic complications. Current evidence suggests that C-peptide replacement therapy might prevent and ameliorate diabetic vasculopathy and organ-specific complications through conservation of vascular function, as well as prevention of endothelial cell death, microvascular permeability, vascular inflammation, and neointima formation. In this review, we describe recent advances on the beneficial role of C-peptide replacement therapy for preventing diabetic complications, such as retinopathy, nephropathy, neuropathy, impaired wound healing, and inflammation, and further discuss potential beneficial effects of combined C-peptide and insulin supplement therapy to control hyperglycaemia and to prevent organ-specific complications.</P>

Characterization of TAMRA- and biotin-conjugated peptide arrays for on-chip matrix metalloproteinase activity assay

공덕훈,Mahendra Prasad Bhatt,이승택,김영명,하권수 한국바이오칩학회 2012 BioChip Journal Vol.6 No.4

Peptide arrays have been widely used for high-throughput determination of protease activities in cells and tissues because specific peptides have high binding affinity for the active site of enzymes. Designing peptide substrate probes for enzyme activity assays have been considered to be important; however, the significance of its reporter tag for detecting enzymatic reactions is relatively underestimated. Thus, we investigated the effect of the reporter tag of peptide substrate probes on on-chip protease activity assays. We optimized and characterized proteolytic activity assay of matrix metalloproteinase-3 using direct and indirect substrate probes, tetramethyl-6-carboxyrhodamine (TAMRA)- and biotin-conjugated peptide arrays, respectively. Proteolytic activity assays using both substrate probes demonstrated similar sensitivity, ratio of maximal to minimal FI, IC50 of GM6001, and interarray reproducibility. However, biotin-conjugated substrate arrays showed a wider dynamic range than TAMRA-conjugated substrate arrays. Thus, this comparative study provides a wealth of information for developing optimal probes necessary for effective analysis of enzyme activity and kinetics.

Prevention of VEGF-mediated microvascular permeability by C-peptide in diabetic mice

Lim, Young-Cheol,Bhatt, Mahendra Prasad,Kwon, Mi-Hye,Park, Donghyun,Lee, Seungkoo,Choe, Jongseon,Hwang, JongYun,Kim, Young-Myeong,Ha, Kwon-Soo Oxford University Press 2014 Cardiovascular research Vol.101 No.1

<P><B>Aims</B></P><P>Human C-peptide has a beneficial effect on the prevention of diabetic neuropathy, nephropathy, and vascular complications; however, its role in protection against increased vascular permeability in diabetes remains unclear. Our purpose was to explore the potential protective role of C-peptide against microvascular permeability mediated by vascular endothelial growth factor (VEGF)-induced reactive oxygen species (ROS) generation in diabetes.</P><P><B>Methods and results</B></P><P>Generation of intracellular ROS, real-time changes in intracellular Ca<SUP>2+</SUP>, ROS-dependent stress fibre formation, and the disassembly of the adherens junctions were studied by a confocal microscopy in human umbilical vein endothelial cells (HUVECs). VEGF-induced vascular leakage was investigated in the skin of diabetic mice using a Miles vascular permeability assay. Microvascular leakage in the retina of streptozotocin diabetic mice was investigated using a confocal microscopy after left ventricle injection of fluorescein isothiocyanate (FITC)-dextran. C-peptide inhibited the VEGF-induced ROS generation, stress fibre formation, disassembly of vascular endothelial cadherin, and endothelial permeability in HUVECs. Intradermal injection of C-peptide prevented VEGF-induced vascular leakage. Consistent with this, intravitreal injection of C-peptide prevented the extravasation of FITC-dextran in the retinas of diabetic mice, which was also prevented by anti-VEGF antibody and ROS scavengers in diabetic mice.</P><P><B>Conclusions/interpretation</B></P><P>C-peptide prevents VEGF-induced microvascular permeability by inhibiting ROS-mediated intracellular events in diabetic mice, suggesting that C-peptide replacement is a promising therapeutic strategy to prevent diabetic retinopathy.</P>

Hidden Risk of Liver Disease in Type 2 Diabetes Population

( Sushant Pokhrel ),( Naresh Pokhrel ),( Bashu Dev Pardhe ),( Anit Lamichhane ),( Rakesh Pokhrel ),( Mahendra Prasad Bhatt ) 대한간학회 2020 춘·추계 학술대회 (KASL) Vol.2020 No.1

Aims: This study aimed to find the association of liver biomarker with diabetes population. Also, this study focused to find out the risk factors associated with liver disease in Nepalese diabetes patient. Methods: This study was carried out among 238 diabetes and 238 apparently healthy population who attended Modern diagnostic laboratory and Research center, Kathmandu, Nepal. HbA1c and fasting plasma glucose was measure to assess the diabetes population and glycemic control. Aspartate to platelet ratio index (APRI), gamma-glutamyl transpeptidase to platelet ratio (GPR), fibrosis-4 index (FIB-4), and triglyceride and glucose index (TyG) were assess for prediction of hidden risk liver disease. Diabetic patient with higher than the cut-off value obtained from ROC curve analysis of different liver marker index were subjected to multivariate regression analysis to measure the independent risk factor for progression liver disease in different model. Results: Patient with poor glycemic control had a significantly higher level of APRI (P=0.05), GPR (P=0.039), and TyG (P<0.001). Higher HbA1c showed significantly positive correlation with APRI (r=0.154, P=0.017), GPR (r=0.203, P=0.002), FIB4 (r=0.132, P=0.042), and TyG (r=0.510, P<0.001) in diabetic population. The Area under ROC curve of GPR was 0.700 (0.654-0.747), APRI 0.839 (0.803-0.874), FIB-4 0.820 (0.783-0.857), and TyG 0.909 (0.882-0.874) with p-value <0.05. The cut-off value (sensitivity, specificity) of GPR was 0.227 (63.4%, 63%), APRI 0.241 (71.0%, 80.3%), FIB-4 1.65 (71.8%, 77.3%), and TyG 8.85 (79%, 93.3%) respectively. Triglyceride, AST, and GGT was independent risk factor followed same trend in different 4 model while HbA1c and ALT showed independent risk factor in 3 models. Conclusions: APRI, GPR, FIB4 and TyG can define the hidden risk liver disease in T2DM. The independent risk factors for progression of liver disease in those population are hypertriglyceridemia, higher AST, and higher GGT. Routinely screening for markers may prevent progression of liver disease in T2DM patients.