Hot Water Extracted Lycium Barbarum and Rehmannia Glutinosa Inhibit Liver Inflammation and Fibrosis in Rats

Polysaccharide-rich Lycium barbarum and Rehmannia glutinosa have been considered to have immune-modulating activity. This study investigated the effects of water extracted Lycium barbarum and Rehmannia glutinosa (HE) on carbon tetrachloride ( CCl4 )-induced liver injury in rats. Male Sprague-Dawley rats were randomly divided into: normal diet + peritoneal injection of olive oil (control), normal diet + CCl4 injection ( CCl4 ), 1 × HE (0.05% HE for each) + CCl4 (1 × HE), and 3 × HE (0.15% HE for each) + CCl4 (3 × HE) groups. Rats were injected with 40% CCl4 at a dose of 0.75 ml/kg body weight once a week for seven weeks, one week after herbal extract treatment. After eight week herbal extract treatment, pathohistological examination showed that both 1× and 3 × HE treatments diminished necrotic hepatocytes, chemoattraction of inflammatory cells, and liver fibrosis. Both 1× and 3 × HE treatments decreased plasma alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities, and reduced hepatic levels of pro-inflammatory cytokines — tumor necrosis factor-α and interleukin-1β — compared to CCl4 treatment alone. The 1 × HE treatment increased hepatic anti-inflammatory cytokine IL-10 levels. Both the 1× and 3 × HE treatments suppressed liver fibrosis biomarkers — transforming growth factor-β1 and hydroxyproline. Therefore, treatment with water extracted Lycium barbarum and Rehmannia glutinosa (0.05% and 0.15% for each) for eight weeks protects against necrotic damage, indicated by decreases in plasma ALT and AST activities, and suppresses liver fibrosis by down-regulation of liver inflammation in rats with CCl4-induced liver injury.

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Targeting ER protein TXNDC5 in hepatic stellate cell mitigates liver fibrosis by repressing non-canonical TGFβ signalling

Gut ◽

10.1136/gutjnl-2021-325065 ◽

2021 ◽

pp. gutjnl-2021-325065

Author(s):

Chen-Ting Hung ◽

Tung-Hung Su ◽

Yen-Ting Chen ◽

Yueh-Feng Wu ◽

You-Tzung Chen ◽

...

Keyword(s):

Extracellular Matrix ◽

Liver Fibrosis ◽

Liver Injury ◽

Transforming Growth Factor ◽

Stellate Cell ◽

Transforming Growth Factor Β1 ◽

Duct Ligation ◽

Extracellular Matrix Production ◽

Matrix Production

Background and objectivesLiver fibrosis (LF) occurs following chronic liver injuries. Currently, there is no effective therapy for LF. Recently, we identified thioredoxin domain containing 5 (TXNDC5), an ER protein disulfide isomerase (PDI), as a critical mediator of cardiac and lung fibrosis. We aimed to determine if TXNDC5 also contributes to LF and its potential as a therapeutic target for LF.DesignHistological and transcriptome analyses on human cirrhotic livers were performed. Col1a1-GFPTg, Alb-Cre;Rosa26-tdTomato and Tie2-Cre/ERT2;Rosa26-tdTomato mice were used to determine the cell type(s) where TXNDC5 was induced following liver injury. In vitro investigations were conducted in human hepatic stellate cells (HSCs). Col1a2-Cre/ERT2;Txndc5fl/fl (Txndc5cKO) and Alb-Cre;Txndc5fl/fl (Txndc5Hep-cKO) mice were generated to delete TXNDC5 in HSCs and hepatocytes, respectively. Carbon tetrachloride treatment and bile duct ligation surgery were employed to induce liver injury/fibrosis in mice. The extent of LF was quantified using histological, imaging and biochemical analyses.ResultsTXNDC5 was upregulated markedly in human and mouse fibrotic livers, particularly in activated HSC at the fibrotic foci. TXNDC5 was induced by transforming growth factor β1 (TGFβ1) in HSCs and it was both required and sufficient for the activation, proliferation, survival and extracellular matrix production of HSC. Mechanistically, TGFβ1 induces TXNDC5 expression through increased ER stress and ATF6-mediated transcriptional regulation. In addition, TXNDC5 promotes LF by redox-dependent JNK and signal transducer and activator of transcription 3 activation in HSCs through its PDI activity, activating HSCs and making them resistant to apoptosis. HSC-specific deletion of Txndc5 reverted established LF in mice.ConclusionsER protein TXNDC5 promotes LF through redox-dependent HSC activation, proliferation and excessive extracellular matrix production. Targeting TXNDC5, therefore, could be a potential novel therapeutic strategy to ameliorate LF.

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

The Scientific World JOURNAL ◽

10.1155/2014/301879 ◽

2014 ◽

Vol 2014 ◽

pp. 1-12 ◽

Cited By ~ 16

Author(s):

Farkaad A. Kadir ◽

Normadiah M. Kassim ◽

Mahmood Ameen Abdulla ◽

Behnam Kamalidehghan ◽

Fatemeh Ahmadipour ◽

...

Keyword(s):

Liver Fibrosis ◽

Transforming Growth Factor ◽

Antioxidant Activities ◽

Smooth Muscle Actin ◽

Immunohistochemical Analysis ◽

Hepatoprotective Activity ◽

Nuclear Antigen ◽

Sprague Dawley ◽

Liver Fibrogenesis

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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Sedum sarmentosum Total Flavonoids Alleviate Schistosomiasis-Induced Liver Fibrosis by Altering TGF-β1 and Smad7 Expression

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2020/2083697 ◽

2020 ◽

Vol 2020 ◽

pp. 1-9

Author(s):

Peng-chun Yang ◽

Wei-zhe Bai ◽

Jing Wang ◽

Cai-hua Yan ◽

Wei-feng Huang ◽

...

Keyword(s):

Liver Fibrosis ◽

Protein Expression ◽

Hepatic Fibrosis ◽

Transforming Growth Factor Beta ◽

Transforming Growth Factor ◽

Schistosomiasis Japonica ◽

Mrna Levels ◽

Sprague Dawley ◽

Tgf Β1 ◽

Liver Tissues

Objectives. Schistosomiasis is a parasitic disease that affects over 142 million people worldwide. The main causes of death of schistosomiasis include liver granuloma and secondary hepatic cirrhosis resulting from severe fibrosis. Despite intensive research, controlling liver fibrosis associated with schistosomiasis remains challenging. Sedum sarmentosum total flavonoid (SSTF) is a promising agent to reduce liver fibrosis with an unknown mechanism. Thus, the objectives of this study are to validate its effect on the liver fibrosis caused by schistosomiasis and to explore the underlying molecular mechanism. Methods. Sixty male Sprague-Dawley rats were randomly divided into six groups: one group of normal control and five groups of liver fibrosis induced by schistosomiasis japonica with or without SSTF or colchicine treatment, the latter serving as the positive control. Liver tissues from each animal were harvested to observe the degree and grade of hepatic fibrosis. We also measured the expression of transforming growth factor-beta 1 (TGF-β1) and Smad7 using RT-qPCR, Western blot, and immunohistochemistry. Results. Compared with the untreated model group, groups treated with SSTF at all three tested doses had significantly reduced hepatic fibrosis ( P < 0.05 ). Each dose of SSTF also significantly reduced TGF-β1 protein expression and mRNA levels in the liver tissues ( P < 0.05 ). In contrast, the middle and high doses of SSTF significantly increased Smad7 protein expression and mRNA levels ( P < 0.05 ). Immunohistochemistry showed that each dose of SSTF reduced TGF-β1 protein expression ( P < 0.05 ). Conclusion. Our results demonstrated that SSTF alleviated schistosomiasis japonica-induced hepatic fibrosis by inhibiting the TGF-β1/Smad7 pathway.

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Oral Astaxanthin Supplementation Prevents Peritoneal Fibrosis in Rats

Peritoneal Dialysis International ◽

10.3747/pdi.2013.00317 ◽

2015 ◽

Vol 35 (5) ◽

pp. 506-516 ◽

Cited By ~ 7

Author(s):

Keiichi Wakabayashi ◽

Chieko Hamada ◽

Reo Kanda ◽

Takanori Nakano ◽

Hiroaki Io ◽

...

Keyword(s):

Mrna Expression ◽

Transforming Growth Factor ◽

Smooth Muscle Actin ◽

Normal Diet ◽

Vascular Density ◽

Related Factor ◽

Sprague Dawley ◽

Peritoneal Fibrosis ◽

Group 3 ◽

Group 1

BackgroundPreventing peritoneal damage during peritoneal dialysis is critical. Reactive oxygen species (ROS) have an important role in peritoneal damage; however, few studies have investigated this. We aimed to determine the effects of oral astaxanthin (AST) supplementation in a peritoneal fibrosis (PF) rat model.MethodsThirty-seven Sprague-Dawley rats were divided into 5 groups: Control 1 (fed a normal diet without stimulation), Control 2 (fed an AST-supplemented diet without stimulation), Group 1 (fed a normal diet with 8% chlorhexidine gluconate [CG] stimulation for 3 weeks), Group 2 (fed a 0.06% AST-supplemented diet with CG stimulation), and Group 3 (fed a 0.06% AST-supplemented diet that was initiated 4 weeks before CG stimulation). Peritoneal fibrosis, vascular proliferation, and fibrosis-related factor expression were examined.ResultsPeritoneal thickness was significantly suppressed by AST supplementation. Astaxanthin diminished the number of CD68-, 8-hydroxy-2'–deoxyguanosine (8-OHdG)-, and monocyte chemoattractant protein-1 (MCP-1)-positive cells. Type 3 collagen, tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and MCP-1 mRNA expression was significantly lower in Group 3 than in Group 1. Increased transforming growth factor-β (TGF-β) and Snail mRNA expression, vascular density, and the number of α–smooth muscle actin (α–SMA)-positive cells were also decreased in Group 3.ConclusionAstaxanthin suppressed PF development through the inhibition of inflammation and oxidation in PF rats. It appears that the anti-oxidative agent AST may be useful for the prevention of peritoneal damage.

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α2-Adrenergic Receptor in Liver Fibrosis: Implications for the Adrenoblocker Mesedin

Cells ◽

10.3390/cells9020456 ◽

2020 ◽

Vol 9 (2) ◽

pp. 456 ◽

Cited By ~ 1

Author(s):

Ute A. Schwinghammer ◽

Magda M. Melkonyan ◽

Lilit Hunanyan ◽

Roman Tremmel ◽

Ralf Weiskirchen ◽

...

Keyword(s):

Growth Factor ◽

Liver Fibrosis ◽

Liver Injury ◽

Transforming Growth Factor ◽

Smooth Muscle Actin ◽

Transforming Growth Factor Β ◽

Liver Sinusoidal Endothelial Cells ◽

Mediated Effects ◽

Liver Sinusoids ◽

Endothelial Nitric Oxide

The noradrenergic system is proposed to play a prominent role in the pathogenesis of liver fibrosis. While α1- and β-adrenergic receptors (ARs) are suggested to be involved in a multitude of profibrogenic actions, little is known about α2-AR-mediated effects and their expression pattern during liver fibrosis and cirrhosis. We explored the expression of α2-AR in two models of experimental liver fibrosis. We further evaluated the capacity of the α2-AR blocker mesedin to deactivate hepatic stellate cells (HSCs) and to increase the permeability of human liver sinusoidal endothelial cells (hLSECs). The mRNA of α2a-, α2b-, and α2c-AR subtypes was uniformly upregulated in carbon tetrachloride-treated mice vs the controls, while in bile duct-ligated mice, only α2b-AR increased in response to liver injury. In murine HSCs, mesedin led to a decrease in α-smooth muscle actin, transforming growth factor-β and α2a-AR expression, which was indicated by RT-qPCR, immunocytochemistry, and Western blot analyses. In a hLSEC line, an increased expression of endothelial nitric oxide synthase was detected along with downregulated transforming growth factor-β. In conclusion, we suggest that the α2-AR blockade alleviates the activation of HSCs and may increase the permeability of liver sinusoids during liver injury.

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Melittin attenuates liver injury in thioacetamide-treated mice through modulating inflammation and fibrogenesis

Experimental Biology and Medicine ◽

10.1258/ebm.2011.011127 ◽

2011 ◽

Vol 236 (11) ◽

pp. 1306-1313 ◽

Cited By ~ 26

Author(s):

Ji-Hyun Park ◽

Yoon-Seup Kum ◽

Tae-Im Lee ◽

Soo-Jung Kim ◽

Woo-Ram Lee ◽

...

Keyword(s):

Liver Fibrosis ◽

Liver Injury ◽

Signaling Pathway ◽

Molecular Mechanisms ◽

Liver Inflammation ◽

Amphipathic Peptide ◽

Bioactive Component ◽

Factor Α ◽

Necrosis Factor

Liver fibrosis represents a process of healing and scarring in response to chronic liver injury. Following injury, an acute inflammation response takes place resulting in moderate cell necrosis and extracellular matrix damage. Melittin, the major bioactive component in the venom of honey bee Apis mellifera, is a 26-residue amphipathic peptide with well-known cytolytic, antimicrobial and proinflammatory properties. However, the molecular mechanisms responsible for the anti-inflammatory activity of melittin have not been elucidated in liver fibrosis. We investigated whether melittin ameliorates liver inflammation and fibrosis in thioacetamide (TAA)-induced liver fibrosis. Two groups of mice were treated with TAA (200 mg/L, in drinking water), one of the groups of mice was co-treated with melittin (0.1 mg/kg) for 12 weeks while the other was not. Hepatic stellate cells (HSCs) were cultured with tumor necrosis factor α in the absence or presence of melittin. Melittin suppresses the expression of proinflammatory cytokines through the nuclear factor (NF)- κB signaling pathway. Moreover, melittin reduces the activity of HSCs in vitro, and decreases the expression of fibrotic gene responses in TAA-induced liver fibrosis. Taken together, melittin prevents TAA-induced liver fibrosis by inhibiting liver inflammation and fibrosis, the mechanism of which is the interruption of the NF- κB signaling pathway. These results suggest that melittin could be an effective agent for preventing liver fibrosis.

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Effects and Mechanisms of Acremoniumterricola milleretal mycelium on Liver Fibrosis Induced by Carbon Tetrachloride in Rats

The American Journal of Chinese Medicine ◽

10.1142/s0192415x11009019 ◽

2011 ◽

Vol 39 (03) ◽

pp. 537-550 ◽

Cited By ~ 10

Author(s):

Xiao-Peng Tian ◽

Yan-Yan Yin ◽

Xia Li

Keyword(s):

Carbon Tetrachloride ◽

Liver Fibrosis ◽

Liver Injury ◽

Chinese Herb ◽

Immunoblot Analysis ◽

Hepatoprotective Activity ◽

Protective Effects ◽

Sprague Dawley ◽

Procollagen Type ◽

Tgf Β1

Acremoniumterricola milleretal mycelium (AMM) is one of the most precious traditional Chinese medicines. It has numerous protective effects on organs, and has been used in Chinese herb prescription to treat refractory diseases. Our preliminary studies demonstrated that AMM had hepatoprotective activity in acute liver injury. We further investigated the effects of AMM on liver fibrosis in rats induced by carbon tetrachloride ( CCl 4) and explore its possible mechanisms. The animal model was established by injection with 50% CCl 4 subcutaneously in male Sprague-Dawley rats twice a week for eight weeks. Meanwhile, AMM (175, 350 and 700 mg/kg) was administered intragastrically per day until sacrifice. We found that treatment with AMM (175, 350 and 700 mg/kg) decreased CCl 4-induced elevation of serum transaminase activities, hyaluronic acid, laminin and procollagen type III levels, and contents of hydroxyproline in liver tissues. It also restored the decreased SOD and GSH-Px activities and inhibited the formation of lipid peroxidative products during CCl 4 treatment. Moreover, AMM (350 and 700 mg/kg) decreased the elevation of TGF-β1 by 19.6% and 34.3%, respectively. In the pathological study, liver injury and the formation of liver fibrosis in rates treated by AMM were improved significantly. Immunoblot analysis showed that AMM (175, 350 and 700 mg/kg) inhibited Smad 2/3 phosphorylation, and elevated inhibitor Smad 7 expression. These results suggested that AMM could protect liver damage and inhibit the progression of hepatic fibrosis induced by CCl 4, and its mechanisms might be associated with its ability to scavenge free radicals, decrease the level of TGF-β1 and block TGF-β/Smad signaling pathway.

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Smad7 Deficiency in Myeloid Cells Does Not Affect Liver Injury, Inflammation or Fibrosis after Chronic CCl4 Exposure in Mice

International Journal of Molecular Sciences ◽

10.3390/ijms222111575 ◽

2021 ◽

Vol 22 (21) ◽

pp. 11575

Author(s):

Ludmilla Unrau ◽

Jessica Endig ◽

Diane Goltz ◽

Paulina Sprezyna ◽

Hanna Ulrich ◽

...

Keyword(s):

Liver Injury ◽

Transforming Growth Factor ◽

Elevated Serum ◽

Myeloid Cells ◽

Chronic Liver ◽

Signalling Pathway ◽

Liver Inflammation ◽

Knock Down ◽

Smad Protein ◽

Smad7 Expression

Myeloid cells play an essential role in the maintenance of liver homeostasis, as well as the initiation and termination of innate and adaptive immune responses. In chronic hepatic inflammation, the production of transforming growth factor beta (TGF-β) is pivotal for scarring and fibrosis induction and progression. TGF-β signalling is tightly regulated via the Smad protein family. Smad7 acts as an inhibitor of the TGF-β-signalling pathway, rendering cells that express high levels of it resistant to TGF-β-dependent signal transduction. In hepatocytes, the absence of Smad7 promotes liver fibrosis. Here, we examine whether Smad7 expression in myeloid cells affects the extent of liver inflammation, injury and fibrosis induction during chronic liver inflammation. Using the well-established model of chronic carbon tetrachloride (CCl4)-mediated liver injury, we investigated the role of Smad7 in myeloid cells in LysM-Cre Smadfl/fl mice that harbour a myeloid-specific knock-down of Smad7. We found that the chronic application of CCl4 induces severe liver injury, with elevated serum alanine transaminase (ALT)/aspartate transaminase (AST) levels, centrilobular and periportal necrosis and immune-cell infiltration. However, the myeloid-specific knock-down of Smad7 did not influence these and other parameters in the CCl4-treated animals. In summary, our results suggest that, during long-term application of CCl4, Smad7 expression in myeloid cells and its potential effects on the TGF-β-signalling pathway are dispensable for regulating the extent of chronic liver injury and inflammation.

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Protective Effects of Ginkgo biloba, Panax ginseng, and Schizandra chinensis Extract on Liver Injury in Rats

The American Journal of Chinese Medicine ◽

10.1142/s0192415x07005466 ◽

2007 ◽

Vol 35 (06) ◽

pp. 995-1009 ◽

Cited By ~ 25

Author(s):

Hsin-Fang Chang ◽

Yun-Ho Lin ◽

Chia-Chou Chu ◽

Shu-Ju Wu ◽

Ya-Hui Tsai ◽

...

Keyword(s):

Liver Fibrosis ◽

Liver Injury ◽

Panax Ginseng ◽

Ginkgo Biloba ◽

Antioxidant Status ◽

Low Dose ◽

Herbal Extract ◽

High Dose ◽

Schizandra Chinensis ◽

Tgf Β1

This study investigated the effects of the combined extracts of Ginkgo biloba, Panax ginseng, and Schizandra chinensis at different doses on hepatic antioxidant status and fibrosis in rats with carbon tetrachloride ( CCl 4)-induced liver injury. Male Sprague-Dawley rats ( n = 8–12 per group) were divided into the control, CCl 4, CCl 4 + silymarin (0.35%), CCl 4 + low-dose herbal extract (0.24% of Ginkgo biloba, Panax ginseng, and Schizandra chinensis extract at 1:1:1; LE), and CCl 4 + high-dose herbal extract (1.20% of the same herbal extract; HE) groups. Silymarin or herbal extract was orally given to rats a week before chronic intraperitoneal injection with CCl 4 for 6 weeks. The pathological results showed that herbal extract suppressed hepatic bile duct proliferation, and low-dose herbal extract inhibited liver fibrosis. Hepatic superoxide dismutase (SOD) activity was lower in the CCl 4 group, but there was no difference in the silymarin or herbal extract treated groups compared to the control group. Hepatic catalase activity and the ratio of reduced to oxidized glutathione were significantly higher ( p < 0.05) in the HE group than those in the CCl 4 group. Silymarin and herbal extract reversed the impaired hepatic total antioxidant status ( p < 0.05). Herbal extract partially reduced the elevated hepatic lipid peroxides. Hepatic transforming growth factor-β1 (TGF-β1) level decreased significantly ( p < 0.05) in the LE group. Therefore, high-dose herbal extract improved hepatic antioxidant capacity through enhancing catalase activity and glutathione redox status, whereas low-dose herbal extract inhibited liver fibrosis through decreasing hepatic TGF-β1 level in rats with CCl 4-induced liver injury.

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Liver fibrosis: a balance of ACEs?

Clinical Science ◽

10.1042/cs20070026 ◽

2007 ◽

Vol 113 (3) ◽

pp. 109-118 ◽

Cited By ~ 68

Author(s):

Fiona J. Warner ◽

John S. Lubel ◽

Geoffrey W. McCaughan ◽

Peter W. Angus

Keyword(s):

Angiotensin Ii ◽

Growth Factor ◽

Liver Fibrosis ◽

Liver Injury ◽

Angiotensin Converting Enzyme ◽

Transforming Growth Factor ◽

Tissue Growth ◽

Chronic Liver ◽

Converting Enzyme ◽

Chronic Liver Injury

There is an increasing body of evidence to suggest that the RAS (renin–angiotensin system) contributes to tissue injury and fibrosis in chronic liver disease. A number of studies have shown that components of a local hepatic RAS are up-regulated in fibrotic livers of humans and in experimental animal models. Angiotensin II, the main physiological effector molecule of this system, mediates liver fibrosis by stimulating fibroblast proliferation (myofibroblast and hepatic stellate cells), infiltration of inflammatory cells, and the release of inflammatory cytokines and growth factors such as TGF (transforming growth factor)-β1, IL (interleukin)-1β, MCP (monocyte chemoattractant protein)-1 and connective tissue growth factor. Furthermore, blockade of the RAS by ACE (angiotensin-converting enzyme) inhibitors and angiotensin type 1 receptor antagonists significantly attenuate liver fibrosis in experimental models of chronic liver injury. In 2000 ACE2 (angiotensin-converting enzyme 2), a human homologue of ACE, was identified. ACE2 efficiently degrades angiotensin II to angiotensin-(1–7), a peptide which has recently been shown to have both vasodilatory and tissue protective effects. This suggests that ACE2 and its products may be part of an alternate enzymatic pathway in the RAS, which counterbalances the generation and actions of angiotensin II, the ACE2–angiotensin-(1–7)–Mas axis. This review focuses on the potential roles of the RAS, angiotensin II and ACE2 in chronic liver injury and fibrogenesis.

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