PASS-Predicted Hepatoprotective Activity ofCaesalpinia sappanin Thioacetamide-Induced Liver Fibrosis in Rats

The antifibrotic effects of traditional medicinal herbCaesalpinia sappan(CS) extract on liver fibrosis induced by thioacetamide (TAA) and the expression of transforming growth factorβ1 (TGF-β1),α-smooth muscle actin (αSMA), and proliferating cell nuclear antigen (PCNA) in rats were studied. A computer-aided prediction of antioxidant and hepatoprotective activities was primarily performed with the Prediction Activity Spectra of the Substance (PASS) Program. Liver fibrosis was induced in male Sprague Dawley rats by TAA administration (0.03% w/v) in drinking water for a period of 12 weeks. Rats were divided into seven groups: control, TAA, Silymarin (SY), and CS 300 mg/kg body weight and 100 mg/kg groups. The effect of CS on liver fibrogenesis was determined by Masson’s trichrome staining, immunohistochemical analysis, and western blotting.In vivodetermination of hepatic antioxidant activities, cytochrome P450 2E1 (CYP2E1), and matrix metalloproteinases (MPPS) was employed. CS treatment had significantly increased hepatic antioxidant enzymes activity in the TAA-treated rats. Liver fibrosis was greatly alleviated in rats when treated with CS extract. CS treatment was noted to normalize the expression of TGF-β1,αSMA, PCNA, MMPs, and TIMP1 proteins. PASS-predicted plant activity could efficiently guide in selecting a promising pharmaceutical lead with high accuracy and required antioxidant and hepatoprotective properties.

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The antioxidant (−)-epigallocatechin-3-gallate inhibits activated hepatic stellate cell growth and suppresses acetaldehyde-induced gene expression

Biochemical Journal ◽

10.1042/bj20020894 ◽

2002 ◽

Vol 368 (3) ◽

pp. 695-704 ◽

Cited By ~ 66

Author(s):

Anping CHEN ◽

Li ZHANG ◽

Jianye XU ◽

Jun TANG

Keyword(s):

Transforming Growth Factor ◽

Stellate Cell ◽

Present Report ◽

Smooth Muscle Actin ◽

Critical Role ◽

Dependent Manner ◽

Liver Fibrogenesis ◽

Underlying Mechanisms

Activated hepatic stellate cells (HSC) are the primary source of excessive production of extracellular matrix during liver fibrogenesis. Although the underlying mechanisms remain incompletely understood, it is widely accepted that oxidative stress plays a critical role in liver fibrogenesis. Suppression of HSC growth and activation, as well as induction of apoptosis, have been proposed as therapeutic strategies for treatment and prevention of this disease. In the present report, we elucidated, for the first time, effects of the antioxidant (—)-epigallocatechin-3-gallate (EGCG), a major (and the most active) component of green tea extracts, on cultured HSC growth and activation. Our results revealed that EGCG significantly inhibited cultured HSC growth by inducing cell cycle arrest and apoptosis in a dose- and time-dependent manner. In addition, EGCG markedly suppressed the activation of cultured HSC as demonstrated by blocking transforming growth factor-β signal transduction and by inhibiting the expression of α1(I) collagen, fibronectin and α-smooth muscle actin genes induced by acetaldehyde, the most active metabolite of ethanol. Furthermore, EGCG reacted differently in the inhibition of nuclear factor-κB activity between cultured HSC with or without acetaldehyde stimulation. Taken together, our results indicated that EGCG was a novel and effective inhibitor for activated HSC growth and activation in vitro. Further studies are necessary to evaluate the effect of this polyphenol in prevention of quiescent HSC activation in vivo, and to further elucidate the underlying mechanisms.

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Mechanism of Hepatoprotective Effect ofBoesenbergia rotundain Thioacetamide-Induced Liver Damage in Rats

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2013/157456 ◽

2013 ◽

Vol 2013 ◽

pp. 1-13 ◽

Cited By ~ 20

Author(s):

Suzy M. Salama ◽

Mahmood A. Abdulla ◽

Ahmed S. AlRashdi ◽

A. Hamid A. Hadi

Keyword(s):

Liver Cirrhosis ◽

Transforming Growth Factor ◽

Ethanolic Extract ◽

Normal Liver ◽

Serum Levels ◽

Hepatoprotective Activity ◽

Tissue Inhibitor Of Metalloproteinase ◽

Nuclear Antigen ◽

Sprague Dawley ◽

Reference Drug

Background. Researchers focused on developing traditional therapies as pharmacological medicines to treat liver cirrhosis.Objectives. Evaluating the hepatoprotective activity ofBoesenbergia rotunda(BR) rhizome ethanolic extract on thioacetamide-induced liver cirrhosis in rats.Methods. MaleSprague-Dawleyrats were intraperitoneally injected with 200 mg/kg TAA 3 times/week and daily oral administration of 250 mg/kg, 500 mg/kg of BR extract, and 50 mg/kg of the reference drug Silymarin for 8 weeks. At the end of the experiment, Masson’s trichrome staining was used to measure the degree of liver fibrosis. Hepatic antioxidant enzymes (CAT and GPx), nitrotyrosine, cytochrome (P450 2E1), matrix metalloproteinase (MMP-2 and MMP-9), tissue inhibitor of metalloproteinase (TIMP-1), and urinary 8-hydroxyguanosine were measured. Serum levels of transforming growth factor TGF-β1, nuclear transcription factor NF-κB, proinflammatory cytokine IL-6, and caspase-3 were evaluated. Serum protein expression and immunohistochemistry of proapoptotic Bax and antiapoptotic Bcl-2 proteins were measured and confirmed by immunohistochemistry of Bax, Bcl-2, and proliferating cell nuclear antigen (PCNA).Results. BR treatment improved liver histopathology, immunohistochemistry, and biochemistry, triggered apoptosis, and inhibited cytokines, extracellular matrix proteins, and hepatocytes proliferation.Conclusion. Liver cirrhosis progression can be inhibited by the antioxidant and anti-inflammatory activities of BR ethanolic extract while preserving the normal liver status.

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Galectin-3 modulates phagocytosis-induced stellate cell activation and liver fibrosis in vivo

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.00257.2011 ◽

2012 ◽

Vol 302 (4) ◽

pp. G439-G446 ◽

Cited By ~ 32

Author(s):

Joy X. Jiang ◽

Xiangling Chen ◽

Daniel K. Hsu ◽

Kornelia Baghy ◽

Nobuko Serizawa ◽

...

Keyword(s):

Liver Fibrosis ◽

Cell Activation ◽

Transforming Growth Factor ◽

Stellate Cell ◽

Smooth Muscle Actin ◽

In Vivo Studies ◽

Enzyme Linked Immunosorbent Assay ◽

Apoptotic Cells ◽

Galectin 3

Hepatic stellate cells (HSC), the key fibrogenic cells of the liver, transdifferentiate into myofibroblasts upon phagocytosis of apoptotic hepatocytes. Galectin-3, a β-galactoside-binding lectin, is a regulator of the phagocytic process. In this study, our aim was to study the mechanism by which extracellular galectin-3 modulates HSC phagocytosis and activation. The role of galectin-3 in engulfment was evaluated by phagocytosis and integrin binding assays in primary HSC. Galectin-3 expression was studied by real-time PCR and enzyme-linked immunosorbent assay, and in vivo studies were done in wild-type and galectin-3−/− mice. We found that HSC from galectin-3−/− mice displayed decreased phagocytic activity, expression of transforming growth factor-β1, and procollagen α1(I). Recombinant galectin-3 reversed this defect, suggesting that extracellular galectin-3 is required for HSC activation. Galectin-3 facilitated the αvβ3 heterodimer-dependent binding, indicating that galectin-3 modulates HSC phagocytosis via cross-linking this integrin and enhancing the tethering of apoptotic cells. Blocking integrin αvβ3 resulted in decreased phagocytosis. Galectin-3 expression and release were induced in active HSC engulfing apoptotic cells, and this was mediated by the nuclear factor-κB signaling. The upregulation of galectin-3 in active HSC was further confirmed in vivo in bile duct-ligated (BDL) rats. Galectin-3−/− mice displayed significantly decreased fibrosis, with reduced expression of α-smooth muscle actin and procollagen α1(I) following BDL. In summary, extracellular galectin-3 plays a key role in liver fibrosis by mediating HSC phagocytosis, activation, and subsequent autocrine and paracrine signaling by a feedforward mechanism.

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Acupuncture Combined with Curcumin Attenuates Carbon Tetrachloride-Induced Hepatic Fibrosis in Rats

Acupuncture in Medicine ◽

10.1136/acupmed-2011-010116 ◽

2012 ◽

Vol 30 (2) ◽

pp. 132-138 ◽

Cited By ~ 6

Author(s):

Feng Zhang ◽

Jin Ma ◽

Yin Lu ◽

Guang-Xia Ni ◽

Chun-Yan Ni ◽

...

Keyword(s):

Carbon Tetrachloride ◽

Liver Fibrosis ◽

Hepatic Fibrosis ◽

Smooth Muscle Actin ◽

Sham Acupuncture ◽

Male Rats ◽

Sprague Dawley ◽

Combined Use ◽

Serum Aspartate Aminotransferase

Background Increasingly, studies demonstrate the effectiveness of acupuncture therapy against liver fibrosis. Curcumin is a natural product with antifibrotic effects, but has poor pharmacokinetic profiles. This study aimed to evaluate whether acupuncture combined with curcumin could more potently attenuate liver fibrosis in chemical intoxicated rats. Methods 60 Sprague–Dawley male rats were randomly divided into control, model, sham, acupuncture, curcumin and combination therapy groups. During the establishment of fibrosis using carbon tetrachloride (CCl4), acupuncture at LR3, LR14, BL18 and ST36 and/or curcumin treatment by mouth were performed simultaneously. After treatment, pathological indexes and histology for hepatic injury and fibrogenesis were detected. The expression of extracellular matrix (ECM) components was also determined. Results Acupuncture combined with curcumin potently protected the liver from CCl4-induced injury and fibrogenesis, as indicated by reduced levels of serum aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, hyaluronic acid, laminin and procollagen III. Combined use also led to significant liver histological improvements. Furthermore, combined use effectively inhibited ECM expression such as α-smooth muscle actin, fibronectin and α1(1) collagen. Conclusions Acupuncture treatment could significantly enhance the antifibrotic efficacy of curcumin on CCl4-induced hepatic fibrosis in rats in vivo, suggesting that a combination of acupuncture with curcumin may be exploited for the prevention of hepatic fibrosis.

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Structure–Activity Relationship of Aloperine Derivatives as New Anti–Liver Fibrogenic Agents

Molecules ◽

10.3390/molecules25214977 ◽

2020 ◽

Vol 25 (21) ◽

pp. 4977

Author(s):

Kun Wang ◽

Zhihao Guo ◽

Yunyang Bao ◽

Yudong Pang ◽

Yinghong Li ◽

...

Keyword(s):

Transforming Growth Factor ◽

Smooth Muscle Actin ◽

Structure Activity Relationship ◽

Transforming Growth Factor Β1 ◽

Activity Relationship ◽

Protein Levels ◽

Structure Activity ◽

Liver Fibrogenesis ◽

Ld50 Value

Twenty-seven novel 12N-substituted aloperine derivatives were synthesized and investigated for their inhibitory effects on collagen α1 (I) (COL1A1) promotor in human hepatic stellate LX-2 cells, taking aloperine (1) as the hit. A structure-activity relationship (SAR) study disclosed that the introduction of suitable substituents on the 12N atom might enhance the activity. Compound 4p exhibited a good promise on down-regulating COL1A1 expression with the IC50 value of 16.5 μM. Its inhibitory activity against COL1A1 was further confirmed on both mRNA and protein levels. Meanwhile, it effectively inhibited the expression of other fibrogenic proteins, such as transforming growth factor β1 (TGF-β1) and smooth muscle actin (α-SMA). It also exhibited good in vivo safety profile with the oral LD50 value of 400 mg kg−1 in mice. The results initiated the anti-liver fibrogenic study of aloperine derivatives, and the key compound 4p was selected as a novel lead for further investigation against liver fibrogenesis.

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Antifibrotic Effect of Marine Ovothiol in an In Vivo Model of Liver Fibrosis

Oxidative Medicine and Cellular Longevity ◽

10.1155/2018/5045734 ◽

2018 ◽

Vol 2018 ◽

pp. 1-10 ◽

Cited By ~ 8

Author(s):

Mariarita Brancaccio ◽

Giuseppe D’Argenio ◽

Vincenzo Lembo ◽

Anna Palumbo ◽

Immacolata Castellano

Keyword(s):

Liver Fibrosis ◽

Matrix Protein ◽

Molecular Mechanisms ◽

Transforming Growth Factor ◽

Smooth Muscle Actin ◽

Extracellular Matrix Protein ◽

In Vivo Model ◽

Protein Accumulation ◽

Antifibrotic Effect

Liver fibrosis is a complex process caused by chronic hepatic injury, which leads to an excessive increase in extracellular matrix protein accumulation and fibrogenesis. Several natural products, including sulfur-containing compounds, have been investigated for their antifibrotic effects; however, the molecular mechanisms underpinning their action are partially still obscure. In this study, we have investigated for the first time the effect of ovothiol A, π-methyl-5-thiohistidine, isolated from sea urchin eggs on an in vivo murine model of liver fibrosis. Mice were intraperitoneally injected with carbon tetrachloride (CCl4) to induce liver fibrosis and treated with ovothiol A at the dose of 50 mg/kg 3 times a week for 2 months. Treatment with ovothiol A caused a significant reduction of collagen fibers as observed by histopathological changes and serum parameters compared to mice treated with control solution. This antifibrotic effect was associated to the decrease of fibrogenic markers involved in liver fibrosis progression, such as the transforming growth factor (TGF-β), the α-smooth muscle actin (α-SMA), and the tissue metalloproteinases inhibitor (TIMP-1). Finally, we provided evidence that the attenuation of liver fibrosis by ovothiol A treatment can be regulated by the expression and activity of the membrane-bound γ-glutamyl-transpeptidase (GGT), which is a key player in maintaining intracellular redox homoeostasis. Overall, these findings indicate that ovothiol A has significant antifibrotic properties and can be considered as a new marine drug or dietary supplement in potential therapeutic strategies for the treatment of liver fibrosis.

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mTOR inhibitors potentially reduce TGF-β2-induced fibrogenic changes in trabecular meshwork cells

Scientific Reports ◽

10.1038/s41598-021-93580-3 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Nozomi Igarashi ◽

Megumi Honjo ◽

Makoto Aihara

Keyword(s):

Trabecular Meshwork ◽

Transforming Growth Factor ◽

Smooth Muscle Actin ◽

Mtor Inhibitors ◽

In Vivo Model ◽

Type I ◽

Fibrotic Response ◽

Alpha Smooth Muscle Actin

AbstractWe examined the effects of mTOR inhibitors on the fibrotic response induced by transforming growth factor-beta2 (TGF-β2) in cultured human trabecular meshwork (hTM) cells. TGF-β2-induced expression of fibronectin, collagen type I, alpha 1 chain (COL1A1), and alpha-smooth muscle actin (αSMA) in hTM cells was examined in the presence or absence of mTOR inhibitors using quantitative real-time polymerase chain reaction, Western blotting, and immunohistochemistry. The migration rates of hTM cells were examined in the presence of TGF-β2 with or without mTOR inhibitors. An in vitro study showed that the expression of fibronectin, COL1A1, and αSMA was upregulated by TGF-β2 treatment of hTM cells; such upregulation was significantly suppressed by mTOR inhibitors. The inhibitors significantly reduced the migration rate of TGF-β2-stimulated hTM cells. mTOR inhibitors may usefully reduce the fibrotic response of hTM cells and we may have to explore if it is also effective in in vivo model.

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Long-Term Aspartame Administration Leads to Fibrosis, Inflammasome Activation, and Gluconeogenesis Impairment in the Liver of Mice

Biology ◽

10.3390/biology10020082 ◽

2021 ◽

Vol 10 (2) ◽

pp. 82

Author(s):

Isabela A. Finamor ◽

Caroline A. Bressan ◽

Isabel Torres-Cuevas ◽

Sergio Rius-Pérez ◽

Marcelo da Veiga ◽

...

Keyword(s):

Liver Fibrosis ◽

Liver Damage ◽

Transforming Growth Factor ◽

Smooth Muscle Actin ◽

Elevated Serum ◽

Inflammasome Activation ◽

Related Factor ◽

Type I ◽

Artificial Sweetener

Background: Aspartame is an artificial sweetener used in foods and beverages worldwide. However, it is linked to oxidative stress, inflammation, and liver damage through mechanisms that are not fully elucidated yet. This work aimed to investigate the effects of long-term administration of aspartame on the oxidative and inflammatory mechanisms associated with liver fibrosis progression in mice. Methods: Mice were divided into two groups with six animals each: control and aspartame. Aspartame (80 mg/kg, via oral) or vehicle was administrated for 12 weeks. Results: Aspartame caused liver damage and elevated serum transaminase levels. Aspartame also generated liver fibrosis, as evidenced by histology analysis, and pro-fibrotic markers’ upregulation, including transforming growth factor β 1, collagen type I alpha 1, and alpha-smooth muscle actin. Furthermore, aspartame reduced nuclear factor erythroid 2-related factor 2 (Nrf2) activation and enzymatic antioxidant activity and increased lipid peroxidation, which triggered NOD-like receptor containing protein 3 (NLRP3) inflammasome activation and p53 induction. Furthermore, aspartame reduced peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PGC-1α) levels, possibly through p53 activation. This PGC-1α deficiency could be responsible for the changes in lipid profile in serum, total lipid accumulation, and gluconeogenesis impairment in liver, evidenced by the gluconeogenic enzymes’ downregulation, thus causing hypoglycemia. Conclusions: This work provides new insights to understand the mechanisms related to the adverse effects of aspartame on liver tissue.

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Piceatannol increases the expression of hepatocyte growth factor and IL-10 thereby protecting hepatocytes in thioacetamide-induced liver fibrosis

Canadian Journal of Physiology and Pharmacology ◽

10.1139/cjpp-2016-0001 ◽

2016 ◽

Vol 94 (7) ◽

pp. 779-787 ◽

Cited By ~ 6

Author(s):

Hazem Abd-Elgawad ◽

Nashwa Abu-Elsaad ◽

Amr El-Karef ◽

Tarek Ibrahim

Keyword(s):

Growth Factor ◽

Liver Fibrosis ◽

Liver Function ◽

Hepatocyte Growth Factor ◽

Transforming Growth Factor ◽

Histopathological Examination ◽

Smooth Muscle Actin ◽

Interleukin 10 ◽

Hepatocyte Growth ◽

Inflammatory Effect

Piceatannol is a polyphenolic analog of resveratrol that selectively inhibits the non-receptor tyrosine kinase-Syk. This study investigates the potential ability of piceatannol to attenuate liver fibrosis and protect hepatocytes from injury. Thioacetamide was injected in adult male mice (100 mg/kg, i.p., 3 times/week) for 8 weeks. Piceatannol (1 or 5 mg/kg per day) was administered by oral gavage during the last 4 weeks. Liver function biomarkers, tissue malondialdehyde (MDA), cytokeratin-18 (CK18), hepatocyte growth factor (HGF), and interleukin-10 (IL-10) were measured. Necroinflammation, fibrosis, expression of transforming growth factor (TGF)-β1, and α-smooth muscle actin (SMA) were scored by histopathological examination and immunohistochemistry. Obtained results showed ability of piceatannol (1 mg/kg) to restore liver function and reduce inflammation. It significantly (p < 0.001) reduced MDA, CK18, TGF-β1, and α-SMA expression, and increased HGF and IL-10. It can be concluded that piceatannol at low dose can inhibit TGF-β1 induced hepatocytes apoptosis and exerts an anti-inflammatory effect attenuating fibrosis progression.

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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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