Protective Effect of Spore Powder of Antrodia camphorata ATCC 200183 on CCl4-Induced Liver Fibrosis in Mice

Liver fibrosis is a pathological process with intrahepatic diffused deposition of the excess extracellular matrix, which leads to various chronic liver diseases. Drugs with high efficacy and low toxicity for liver fibrosis are still unavailable. Antrodia camphorata has antioxidant, antivirus, antitumor and anti-inflammation roles, and has been used to treat liver diseases in the population. However, the hepatoprotective effects of A. camphorata spores and the mechanisms behind it have not been investigated. In this study, we evaluate the hepatoprotective effect of spore powder of A. camphorata (SP, 100 mg/kg/day or 200 mg/kg/day) on carbon tetrachloride (CCl4)-induced liver fibrosis in mice. SP groups reduced serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities compared with the CCl4 group. SP also showed a decrease in hydroxyproline (Hyp) content in liver tissues. SP improved cell damage and reduced collagen deposition by H&E, Sirius red and Masson staining. Furthermore, SP down-regulated the mRNA levels of α-SMA and Col 1, and the protein expression of α-smooth muscle actin (α-SMA), collagen I (Col 1), tumor necrosis factor alpha (TNF-α), toll like receptor 4 (TLR4) and nuclear factor-Κb (NF-κB) p65. In summary, SP has an ameliorative effect on hepatic fibrosis, probably by inhibiting the activation of hepatic stellate cells, reducing the synthesis of extracellular matrix.

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Inhibition of p300/CBP-Associated Factor Attenuates Renal Tubulointerstitial Fibrosis through Modulation of NF-kB and Nrf2

International Journal of Molecular Sciences ◽

10.3390/ijms20071554 ◽

2019 ◽

Vol 20 (7) ◽

pp. 1554 ◽

Cited By ~ 7

Author(s):

Sungjin Chung ◽

Soojeong Kim ◽

Mina Son ◽

Minyoung Kim ◽

Eun Koh ◽

...

Keyword(s):

Transforming Growth Factor ◽

Cell Damage ◽

Smooth Muscle Actin ◽

Transforming Growth Factor Β ◽

Nuclear Factor Κb ◽

Tubulointerstitial Fibrosis ◽

Mrna Levels ◽

Nuclear Factor ◽

Associated Factor ◽

Renal Tubulointerstitial Fibrosis

p300/CBP-associated factor (PCAF), a histone acetyltransferase, is involved in many cellular processes such as differentiation, proliferation, apoptosis, and reaction to cell damage by modulating the activities of several genes and proteins through the acetylation of either the histones or transcription factors. Here, we examined a pathogenic role of PCAF and its potential as a novel therapeutic target in the progression of renal tubulointerstitial fibrosis induced by non-diabetic unilateral ureteral obstruction (UUO) in male C57BL/6 mice. Administration of garcinol, a PCAF inhibitor, reversed a UUO-induced increase in the renal expression of total PCAF and histone 3 lysine 9 acetylation and reduced positive areas of trichrome and α-smooth muscle actin and collagen content. Treatment with garcinol also decreased mRNA levels of transforming growth factor-β, matrix metalloproteinase (MMP)-2, MMP-9, and fibronectin. Furthermore, garcinol suppressed nuclear factor-κB (NF-κB) and pro-inflammatory cytokines such as tumor necrosis factor-α and IL-6, whereas it preserved the nuclear expression of nuclear factor erythroid-derived 2-like factor 2 (Nrf2) and levels of Nrf2-dependent antioxidants including heme oxygense-1, catalase, superoxide dismutase 1, and NAD(P)H:quinone oxidoreductase 1. These results suggest that the inhibition of inordinately enhanced PCAF could mitigate renal fibrosis by redressing aberrant balance between inflammatory signaling and antioxidant response through the modulation of NF-κB and Nrf2.

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Sestrin 2 Attenuates Rat Hepatic Stellate Cell (HSC) Activation and Liver Fibrosis via an mTOR/AMPK-Dependent Mechanism

Cellular Physiology and Biochemistry ◽

10.1159/000495829 ◽

2018 ◽

Vol 51 (5) ◽

pp. 2111-2122 ◽

Cited By ~ 7

Author(s):

Yi-Bing Hu ◽

Xiao-Ting Ye ◽

Qing-Qing Zhou ◽

Rong-Quan Fu

Keyword(s):

Liver Fibrosis ◽

Protein Expression ◽

Hepatic Fibrosis ◽

Transforming Growth Factor ◽

Histopathological Examination ◽

Stellate Cell ◽

Smooth Muscle Actin ◽

Mrna Levels ◽

Alpha Smooth Muscle Actin

Background/Aims: Sestrin 2 is associated with the pathophysiology of several diseases. The aim of this study was to investigate the effects and potential mechanisms of Sestrin 2 in rat hepatic stellate cells (HSCs) during liver fibrogenesis. Methods: In this study, Sestrin 2 protein expression was detected in rat HSC-T6 cells challenged with transforming growth factor-β (TGF-β) and in mice treated with carbon tetrachloride (CCl4), a well-known model of hepatic fibrosis. Next, HSC-T6 cells and fibrotic mice were transfected with lentivirus. The mRNA expression levels of markers of liver fibrosis [alpha-smooth muscle actin (α-SMA) and collagen 1A1 (Col1A1)] were analyzed by quantitative reverse transcription–polymerase chain reaction (RT-PCR). Cell death and proliferation were evaluated by the MTT assay, and biochemical markers of liver damage in serum [alanine transaminase (ALT) and aspartate transaminase (AST)] were also measured using a biochemical analyzer. Histopathological examination was used to evaluate the degree of liver fibrosis, and protein expression [phospho-adenosine monophosphate-activated protein kinase (p-AMPK), AMPK, phospho-mammalian target of rapamycin (p-mTOR), and mTOR] was determined by western blotting. Results: We found that Sestrin 2 was elevated in both the HSC-T6 cell and hepatic fibrosis models. In vitro, overexpression of Sestrin 2 attenuated the mRNA levels of α-SMA and Col1A1, suppressed α-SMA protein expression, and modulated HSC-T6 cell proliferation. In vivo, overexpression of Sestrin 2 reduced the ALT and AST levels as well as the α-SMA and Col1A1 protein expression in the CCl4 model of liver fibrosis. Moreover, the degree of liver fibrosis was ameliorated. Interestingly, overexpression of Sestrin 2 increased p-AMPK but decreased p-mTOR protein expression. Conclusion: Our findings indicate that Sestrin 2 may attenuate the activation of HSCs and ameliorate liver fibrosis, most likely via upregulation of AMPK phosphorylation and suppression of the mTOR signaling pathway.

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Crocin attenuates CCl4-induced liver fibrosis via PPAR-γ mediated modulation of inflammation and fibrogenesis in rats

Human & Experimental Toxicology ◽

10.1177/0960327120937048 ◽

2020 ◽

Vol 39 (12) ◽

pp. 1639-1649 ◽

Cited By ~ 1

Author(s):

J Chhimwal ◽

S Sharma ◽

P Kulurkar ◽

V Patial

Keyword(s):

Liver Fibrosis ◽

Liver Tissue ◽

Transforming Growth Factor ◽

Liver Enzymes ◽

Pathological Condition ◽

Smooth Muscle Actin ◽

Intraperitoneal Administration ◽

Crocus Sativus ◽

Mrna Levels ◽

Ppar Γ

Background: Liver fibrosis is a chronic pathological condition with a leading cause of liver-related mortality worldwide. In the present study, we have evaluated the antifibrotic effect of crocin, a carotenoid present in the stigma of Crocus sativus, and also explored its putative mechanism of action. Methods: Liver fibrosis was induced by intraperitoneal administration of 30% carbon tetrachloride (CCl4). The crocin was administered orally at 20, 40 and 80 mg/kg body weight along with CCl4 up to 8 weeks. Results: Chronic exposure to CCl4 resulted in elevated levels of liver enzymes and reduced cytochrome P450 2E1 (CYP2E1) activity in the liver. The liver tissue showed cellular swelling, vacuolization, necrosis, infiltration of inflammatory cells and fibrotic changes. The crocin treatment significantly lowered the levels of liver enzymes in serum and improved the liver CYP2E1 mRNA levels. The pathological changes in the liver were also lowered by crocin treatment. The level of pro-inflammatory cytokines, nuclear factor-kappa B, interleukin-6 and tumor necrosis factor α and fibrogenic factor, transforming growth factor β, and α-smooth muscle actin were elevated by the CCl4 in the liver tissue. However, crocin treatment at different doses significantly reduced the expression of these factors. The increased caspase 3/7 activity was also lowered by crocin. CCl4 administration decreased the expression of peroxisome proliferator-activated receptor γ (PPAR-γ) in liver tissue. The improved PPAR-γ expression in the liver by crocin treatment indicates its role in the therapeutic effect of crocin. Conclusions: Crocin attenuated the various events in the progression of liver fibrosis via PPAR-γ mediated modulation of inflammatory and fibrogenic pathways.

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Therapeutic Effect of Dunaliella salina Microalgae on Thioacetamide- (TAA-) Induced Hepatic Liver Fibrosis in Rats: Role of TGF-β and MMP9

BioMed Research International ◽

10.1155/2019/7028314 ◽

2019 ◽

Vol 2019 ◽

pp. 1-9 ◽

Cited By ~ 6

Author(s):

Farouk K. El-Baz ◽

Abeer Salama ◽

Rania A. A. Salama

Keyword(s):

Liver Fibrosis ◽

Transforming Growth Factor Beta ◽

Dunaliella Salina ◽

Transforming Growth Factor ◽

Smooth Muscle Actin ◽

Elevated Serum ◽

Serum Levels ◽

Dependent Manner ◽

Necrosis Factor Alpha ◽

Liver Histopathology

Liver fibrosis represents a serious global health-care problem and is the outcome of many chronic liver diseases, cirrhosis, hepatitis, and toxin accumulation. The present study aimed to evaluate the antifibrotic curative effect of Dunaliella salina (D. salina) on thioacetamide- (TAA-) induced liver fibrosis in rats. Liver fibrosis was induced by TAA (200mg/kg; i.p.) twice per week for 6 weeks. D. salina was given orally (100 and 200 mg/kg) and silymarin was given orally (100 mg/kg) daily, for 4 weeks after TAA. Serum transaminase activities, liver inflammatory cytokines, fibrotic biomarkers, and liver histopathology were assessed. TAA significantly (p<0.05) elevated serum activities of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) and serum levels of total bilirubin, tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and transforming growth factor-beta (TGF-β) with a reduction in albumin. In addition, TAA increased hepatic contents of collagen I, a-smooth muscle actin (α-SMA), and tissue inhibitors of metalloproteinase (TIMP-1), reduced matrix metalloproteinase 9 (MMP9), and finally produced marked degeneration and fibrosis of hepatocytes. Treatment with D. salina or silymarin improved the histological feature of hepatocytes and ameliorated the deleterious effects of TAA in a dose-dependent manner. Based on these results, it could be concluded that the use of D. salina could be assigned for liver fibrosis treatment via its anti-inflammatory and antifibrotic properties.

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Reduced liver fibrosis in hypoxia-inducible factor-1α-deficient mice

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.90368.2008 ◽

2009 ◽

Vol 296 (3) ◽

pp. G582-G592 ◽

Cited By ~ 131

Author(s):

Jeon-OK Moon ◽

Timothy P. Welch ◽

Frank J. Gonzalez ◽

Bryan L. Copple

Keyword(s):

Liver Fibrosis ◽

Type I Collagen ◽

Smooth Muscle Actin ◽

Hypoxia Inducible Factor ◽

Type I ◽

Mrna Levels ◽

Chronic Injury ◽

Hypoxia Inducible Factor 1Α ◽

Duct Ligation ◽

Deficient Mice

Liver fibrosis is characterized by excessive deposition of extracellular matrix in the liver during chronic injury. During early stages of this disease, cells begin to synthesize and secrete profibrotic proteins that stimulate matrix production and inhibit matrix degradation. Although it is clear that these proteins are important for development of fibrosis, what remains unknown is the mechanism by which chronic liver injury stimulates their production. In the present study, the hypothesis was tested that hypoxia-inducible factor-1α (HIF-1α) is activated in the liver during chronic injury and regulates expression of profibrotic proteins. To investigate this hypothesis, mice were subjected to bile duct ligation (BDL), an animal model of liver fibrosis. HIF-1α protein was increased in the livers of mice subjected to BDL by 3 days after surgery. To test the hypothesis that HIF-1α is required for the development of fibrosis, control and HIF-1α-deficient mice were subjected to BDL. Levels of type I collagen and α-smooth muscle actin mRNA and protein were increased in control mice by 14 days after BDL. These levels were significantly reduced in HIF-1α-deficient mice. Next, the levels of several profibrotic mediators were measured to elucidate the mechanism by which HIF-1α promotes liver fibrosis. Platelet-derived growth factor (PDGF)-A, PDGF-B, and plasminogen activator inhibitor-1 mRNA levels were increased to a greater extent in control mice subjected to BDL compared with HIF-1α-deficient mice at 7 and 14 days after BDL. Results from these studies suggest that HIF-1α is a critical regulator of profibrotic mediator production during the development of liver fibrosis.

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Human adipose-derived stem cells pre-treated with platelet-rich plasma and hepatocyte growth factor alleviate liver fibrosis in mice

Biomedical Research and Therapy ◽

10.15419/bmrat.v5i5.446 ◽

2018 ◽

Vol 5 (5) ◽

pp. 2332-2348 ◽

Cited By ~ 1

Author(s):

Nam H. Nguyen ◽

Trinh V. Le ◽

Huy Q. Do ◽

Thanh M. Dang ◽

Yen K. T. Nguyen ◽

...

Keyword(s):

Stem Cells ◽

Growth Factor ◽

Liver Fibrosis ◽

Hepatocyte Growth Factor ◽

Platelet Rich Plasma ◽

Smooth Muscle Actin ◽

Mrna Levels ◽

Adipose Derived Stem Cells ◽

Healthy Human ◽

Hepatocyte Growth

Background: Transplantation of adipose-derived stem cells (ADSCs) is a potential therapy for a variety of liver diseases. Previous studies have shown that ADSC-based therapy is promising for liver fibrosis treatment. Recently, many publications have suggested that pretreating ADSCs with growth factors before transplantation can elevate the effectiveness of the therapy. Therefore, we hypothesize that human ADSCs (hADSCs) pretreated with platelet-rich plasma (PRP) and hepatocyte growth factor (HGF), compared to ADSCs alone, would accelerate the treatment effects on liver fibrosis in mice. Methods: The hADSCs were cultured solely with conventional media, or with HGF (human recombinant; 20 ng/ml), or with HGF and PRP (from healthy human blood, 10%), concomitantly added to the medium for 7 days before transplantation. Eight-week-old male mice were treated with CCl4 (1 ml/kg) for 11 weeks to induce liver fibrosis. The mice were then subsequently divided into: 1) Placebo group (PBS injection); 2) ADSCs/HGF+PRP (5x105HGF and PRP pre-treated cells/mice); 3) ADSCs/HGF (5x105HGF pre-treated cells/mice) and 4) ADSCs (5x105non-pretreated cells/mice). Results: Seven days post-transplantation, the alanine transaminase (ALT) level in the placebo was notably elevated (143.10±14.96 IU/L), compared to ALT levels of ADSCs-, ADSCs/HGF+PR-, and ADSCs/HGF-transplanted mice, which showed an improvement (67.94±18.57 IU/L, 49.44±7.56 IU/L, and 57.93±5.75 IU/L, respectively). The procollagen-α1 and alpha-smooth muscle actin (α-SMA) mRNA levels were significantly down-regulated in the ADSCs-transplanted group compared to those of the placebo. Importantly, these levels were lower in ADSCs/HGF+PR (procollagen-α1: 75.64±45.89; α-SMA: 36.17±36.09) than those of ADSCs/HGF (procollagen-α1: 212.8±84.35; α-SMA: 52.41±7.93) and ADSCs only (procollagen-α1: 310.50 ± 55.36; α-SMA: 184.70±14.06). Stem cell transplantation also improved histological index, reducing inflammation and collagen/necrotic structure accumulation. However, there were no statistical differences between three ADSCs treatment groups after 14 days after transplantation. Conclusion: Pre-treatment with PRP and HGF for 7 days enhanced the efficacy of ADSCs in alleviating liver fibrosis in vivo.

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Orphan nuclear receptor NR4A2 inhibits hepatic stellate cell proliferation through MAPK pathway in liver fibrosis

PeerJ ◽

10.7717/peerj.1518 ◽

2015 ◽

Vol 3 ◽

pp. e1518 ◽

Cited By ~ 10

Author(s):

Pengguo Chen ◽

Jie Li ◽

Yan Huo ◽

Jin Lu ◽

Lili Wan ◽

...

Keyword(s):

Extracellular Matrix ◽

Cell Proliferation ◽

Liver Fibrosis ◽

Nuclear Receptor ◽

Mapk Pathway ◽

Stellate Cell ◽

Smooth Muscle Actin ◽

Control Group ◽

Orphan Nuclear Receptor ◽

Fibrotic Liver

Hepatic stellate cells (HSCs) play a crucial role in liver fibrosis, which is a pathological process characterized by extracellular matrix accumulation. NR4A2 is a nuclear receptor belonging to the NR4A subfamily and vital in regulating cell growth, metabolism, inflammation and other biological functions. However, its role in HSCs is unclear. We analyzed NR4A2 expression in fibrotic liver and stimulated HSCs compared with control group and studied the influence on cell proliferation, cell cycle, cell apoptosis and MAPK pathway after NR4A2 knockdown. NR4A2 expression was examined by real-time polymerase chain reaction, Western blotting, immunohistochemistry and immunofluorescence analyses. NR4A2 expression was significantly lower in fibrotic liver tissues and PDGF BB or TGF-βstimulated HSCs compared with control group. After NR4A2 knockdownα-smooth muscle actin and Col1 expression increased. In addition, NR4A2 silencing led to the promotion of cell proliferation, increase of cell percentage in S phase and reduced phosphorylation of ERK1/2, P38 and JNK in HSCs. These results indicate that NR4A2 can inhibit HSC proliferation through MAPK pathway and decrease extracellular matrix in liver fibrogenesis. NR4A2 may be a promising therapeutic target for liver fibrosis.

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Detection of Pathological Changes in the Aorta during Thoracic Aortic Aneurysm Progression on Molecular Level

Disease Markers ◽

10.1155/2017/9185934 ◽

2017 ◽

Vol 2017 ◽

pp. 1-11 ◽

Cited By ~ 3

Author(s):

Miroslava Rabajdová ◽

Peter Urban ◽

Ivana Špaková ◽

Artemiou Panagiotis ◽

Michaela Ferenčáková ◽

...

Keyword(s):

Gene Expression ◽

Extracellular Matrix ◽

Aortic Aneurysm ◽

Thoracic Aortic Aneurysm ◽

Aortic Wall ◽

Pathological Process ◽

Aortic Tissue ◽

Mrna Levels ◽

Tissue Samples ◽

Extracellular Matrix Components

The progression of thoracic aortic aneurysm depends on regulation of aortic wall homeostasis and on changes in the structural components of the extracellular matrix, which are affected by multiple molecular signalling pathways. We decided to correlate the diameter of ascending thoracic aneurysm with gene expression of inflammation markers (IL-6, CRP), cytokine receptors (IL-6R, TNFR1, and TNFR2), and extracellular matrix components (Emilin-1, MMP9, and TIMP) for detection of the degree of pathological process of TAA formation. The experimental group was divided into three groups according to the diameter of the aortic aneurysm. Whole blood and tissue samples were properly collected and used for nucleic acid, chromatin, and protein isolation. The mRNA levels were detected by qRT-PCR. For the detection of protein levels a Cytokine Array IV assay kit was used in combination with a biochip analyzer. In aortic tissue, significant positive correlations were found between increased mRNA levels of inflammatory cytokines (CRP and IL-6) on both mRNA levels in tissue and protein from the blood with maximum in stage 3. Changes of gene expression of selected genes can be used for the experimental study of the inflammatory receptor inhibitors during trials targeted on slowing down the progress of aortic wall aneurysm.

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Upregulation of Connexin-43 is Critical for Irradiation-induced Neuroinflammation

CNS & Neurological Disorders - Drug Targets ◽

10.2174/1871527317666180706124602 ◽

2018 ◽

Vol 17 (7) ◽

pp. 539-546 ◽

Cited By ~ 1

Author(s):

Wei Chen ◽

Wusong Tong ◽

Yijun Guo ◽

Bin He ◽

Lei Chen ◽

...

Keyword(s):

Ischemic Stroke ◽

Connexin 43 ◽

Interleukin 1 ◽

Pathological Process ◽

Inflammatory Factors ◽

Mrna Levels ◽

Necrosis Factor Alpha ◽

Γ Radiation ◽

Neural Damage ◽

Cx 43

Background: Radiation therapy is widely used for the treatment of pituitary adenomas. Unfortunately, it might raise the risk of ischemic stroke, with neuroinflammation being a major pathological process. Astrocytes are the most abundant cell type in the central nervous system and have been reported for playing important roles in ischemic stroke. Objective: Here we studied how γ-radiation would introduce astrocytes into a detrimental state for neuroinflammation and provide new theory evidence and target for the clinical management of inflammation- related neural damage after radiation-induced ischemic stroke. Method: HA-1800 cells were treated with γ-radiation and then the protein and mRNA levels of Connexin (Cx)-43 were evaluated by western and q-PCR. The culture supernatant was collected and the concentrations of the inflammatory factors were determined by ELISA. MiRNA complementary to Cx-43 was designed through the online tools. Results: Cx-43 is upregulated in the treatment of γ-radiation in astrocytes and γ-radiation introduced the detrimental function of astrocytes: cell viability was reduced while the apoptotic cells were increased. Inflammatory factors like tumor necrosis factor alpha, interferon gamma, interleukin-6, interleukin 1-beta were dramatically up-regulated by the irradiation. MiR-374a rescued irradiation induced Cx-43 up-regulation of astrocytes and eliminated detrimental function triggered by γ-radiation. Conclusion: Cx-43 expression level may play an important role in the inflammation-related neural damage after irradiation-induced ischemic stroke.

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Evaluation of methanolic extract of Phragmites karka on carbon tetrachloride-induced liver fibrosis in rat

Bangladesh Journal of Pharmacology ◽

10.3329/bjp.v12i3.32127 ◽

2017 ◽

Vol 12 (3) ◽

pp. 276 ◽

Cited By ~ 1

Author(s):

Atta Ur Rehman ◽

Manal Liaqat ◽

Rabia Asghar ◽

Nawazish-i-Husain Syed

Keyword(s):

Extracellular Matrix ◽

Liver Fibrosis ◽

Transforming Growth Factor Beta ◽

Transforming Growth Factor ◽

Smooth Muscle Actin ◽

Serum Levels ◽

Matrix Deposition ◽

Male Wistar Rats ◽

Phragmites Karka ◽

Extracellular Matrix Deposition

Phragmites karka has been reported for its anti-inflammatory and analgesic activities. Here, extracts of leaf and rhizome of the plant were individually investigated in CCl4-induced hepatofibrosis in male Wistar rats by administering CCl4 intraperitoneally biweekly for 6 weeks. Afterwards the animals were investigated for liver fibrosis at biochemical, molecular and histological levels, and it showed a profound increase (p<0.001) in elevation of serum levels of transaminases, γ-glutamyl transpeptidase, mRNA expression of α smooth muscle actin, collagen and transforming growth factor beta (TGFβ), and extracellular matrix deposition and perilobular necrosis. Both extracts markedly (p<0.001) decreased the elevated levels of these markers. Histopathological investigations also substantiated the above results by exhibiting a decreased in extracellular matrix deposition in post-treatment animals. In conclusion, both extracts had substantially modified the biochemical and molecular markers of liver fibrosis, in addition to histological improvement in architecture of liver.

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