Liver fibrosis associated with adipose tissue and liver inflammation in an obesity model.

INTRODUCTION: Obesity, the main risk factor for type 2 diabetes mellitus (T2DM), affects the secretion of various hormones that lead to change in metabolism. Visceral adipose tissue accumulation may contribute to Non-alcoholic Fatty Liver Disease (NAFLD) and induce liver injury. This study was aimed to investigate the association between adipose tissue inflammation and liver fibrosis. MATERIALS AND METHODS: Wistar male rats (3 months old, 160- 230 grams) were divided into 4 groups that consisted of six rats in each group. The obesity model was induced through the administration of high-fat diet for a month (OB1), two months (OB2), and four months (OB4). Standard chow was provided for the control group for four months. After the specified date the rats were euthanized and the liver and retroperitoneal white adipose tissue (RWAT) were harvested. We performed RT-PCR to assess the mRNA expressions involved in proinflammatory mediators, fibrosis and antifibrosis signaling. Sirius red staining was performed to assess liver fibrosis. Data were analyzed with SPSS 23 for Windows with significance set as p<0.05. RESULTS: Obesity-induced high-fat diet stimulated an increase of body mass index (BMI) in the OB groups (p<0.05) compared to the control group. Increased BMI was followed by upregulation of proinflammatory mediators (MCP-1, CD68, TLR4, and NFκB) of the RWAT and liver in the obese groups (p<0.05), which promoted hepatic fibrosis in triad portal areas and upregulation of TGFß (p<0.05) mRNA expression as well as downregulation of HGF and c-Met (p<0.05). In addition, hepatic ppET1 and EDNRB mRNA level expressions (p<0.05) were obviously upregulated in the obese groups followed by downregulation of eNOS (p<0.05) mRNA expressions. CONCLUSION: Obesity enhanced inflammation in RWAT and was associated with inflammation and fibrosis of liver.

Upregulation of Megalin, Cubilin, NGAL mRNA expression in kidney may represent tubular injury and apoptosis in chronic condition of rat diabetic model.

INTRODUCTION: Diabetes mellitus (DM) leads to microvascular injury development and produces diabetes nephropathy (DN) with proteinuria, tubular injury, apoptosis and autophagy with upregulation of Bax, BASP and mTORC-1. Megalin, Cubilin and Neutrophil Gelatinase Associated Lipocalin (NGAL) play role in acute pathological condition of kidney injury, however its expression in chronic and slowly progressive kidney injury such as DN has not been elucidated yet. This study focuses upregulation of Megalin, Cubilin and NGAL in association with tubular injury and apoptosis in DN condition. MATERIALS AND METHODS: Diabetic condition was performed with intraperitoneal injection of Streptozotocin 60 mg/kg body weight (BW) in Sprague Dawley rats (2 months old, n=24), and were kept for 1, 2, and 4 months (DM1, DM2, and DM4, respectively). Control group was injected with NaCl 0.9%. Serum glucose level and proteinuria score were assessed, furthermore tubular injury score was quantified based on Periodic-Acid Schiff (PAS) staining. Reverse Transcriptase-PCR (RT-PCR) was carried out for NGAL, Megalin, Cubilin, m-TOR, Bax, and BASP-1 mRNA expression. Data were analyzed using SPSS 22 software. RESULTS: DM led to kidney injury in this model with significant higher glucose level, proteinuria and tubular injury, especially in DM4 group which represented chronic phase of DN and CKD. These findings associated with upregulation of Megalin,Cubilin and NGAL mRNA expression in DM groups, especially in DM4 group. DM4 group also revealed higher expression of Bax, BASP and mTOR mRNA expression which demonstrated apoptosis. CONCLUSION: Megalin, Cubilin and NGAL upregulation may represent tubular injury and apoptosis as progression of DN.

Chlorogenic acid attenuates kidney fibrosis via antifibrotic action of BMP-7 and HGF.

BACKGROUND: Kidney fibrosis, characterised by tubulointerstitial fibrosis, is a histological landmark of chronic kidney disease. The body attempts to compensate for progressive detrimental process of kidney fibrosis by producing antifibrotic substances, such as bone morphogenetic protein-7 (BMP-7) and hepatocyte growth factor (HGF). Chlorogenic acid is known to have renoprotective and antifibrotic properties. This study aims to evaluate the effect of chlorogenic acid on unilateral ureteral obstruction (UUO)-induced kidney fibrosis mice model. METHODS: This study was a quasi-experimental with posttestonly control group design. Twenty-five adult male Swiss Webster mice were randomly divided into five groups: shamoperated group (SO), UUO-control day-7 (U7), UUO-control day-14 (U14), UUO-chlorogenic acid day-7 (UC7), and UUOchlorogenic acid day 14 (UC14). Myofibroblasts were identified by immunohistochemical staining of alphasmooth muscle actin (α-SMA) while collagen fibers were identified by Sirius Red staining. Both data were presented as area fraction. BMP-7 and HGF mRNA expressions were assessed by reverse transcription PCR (RT-PCR). Data were quantified using ImageJ software. RESULTS: UUO-control groups (U7 and U14) showed higher α- SMA-immunopositive (6.52±1.33, 18.24±1.39 vs. 0.22±0.01; p<0.05) and Sirius Red-positive area fractions (6.61±0.8, 12.98±2.31 vs. 0.62±0.10; p<0.05), lower BMP-7 (1.02±0.47, 1.18±0.65 vs. 2.09±0.87; p<0.05) and HGF mRNA expressions (1.06±0.31, 0.89±0.14 vs. 1.88±0.81; p<0.05) compared to SO group. UUO-chlorogenic acid groups (UC7 and UC14) showed lower α-SMA-immunopositive (1.24±0.37, 4.58±0.61; p<0.05) and Sirius Red-positive area fractions (4.76±1.03, 3.72±0.54; p<0.05), higher BMP-7 (1.84±0.49, 2.19±0.43; p<0.05) and HGF (1.58±0.38; p>0.05, 1.84±0.42; p<0.05) mRNA expressions compared to UUO-control groups. UUOchlorogenic acid groups showed BMP-7 and HGF mRNA expressions that were not significantly different from the SO group. CONCLUSION: Chlorogenic acid administration prevents kidney fibrosis in UUO mice model through modulating antifibrotic pathway.

Uric acid induces liver fibrosis through activation of inflammatory mediators and proliferating hepatic stellate cell in mice.

INTRODUCTION: Uric acid is associated with cardiometabolic risk factor and severity of liver damage. The mechanism of uric acid inducing liver damage is still elusive. This study elucidates the development of liver fibrosis under hyperuricemia. METHODS AND MATERIALS: Hyperuricemia model was performed in male Swiss Webster mice. Intraperitoneally injection of uric acid (125mg/kg body weight) was done for 7 and 14 days (UA7 and UA14 groups). Meanwhile, the UAL groups were injected with uric acid and followed by the administration of allopurinol (UAL7 and UAL14 groups). On the due date, mice were sacrificed, and liver was harvested. Uric acid, SGOT, SGPT, and albumin level were measured from the serum. The mRNA expression of TLR4, MCP1, CD68, and collagen1 were assessed through RT-PCR. Liver fibrosis was quantified through Sirius red staining, while the number of hepatic stellates cells (HSCs) and TLR4 were assessed through IHC staining. RESULTS: Uric acid induction for 7 and 14 days stimulated an increase of both SGOT and SGPT serum levels. Followed by enhanced inflammatory mediators: Toll-like receptor-4 (TLR- 4), Monocyte Chemoattractant Protein-1 (MCP-1) and Cluster of Differentiation 68 (CD68) mRNA expression in the liver (p<0.05). The histological findings showed that the UA7 and UA14 groups had higher liver fibrosis scores (p<0.05), collagen I mRNA expression (p<0.05), and the number of HSCs (p<0.05) compared to Control group. Administration of allopurinol showed amelioration of uric acid and liver enzymes levels which followed by inflammatory mediators, liver fibrosis and collagen1, and hepatic stellate cells significantly. CONCLUSION: Therefore, uric acid augmented the liver fibrosis by increasing the number of hepatic stellate cells.