Ginsenosides: potential therapeutic agents against hepatic fibrosis

Hepatology Basic Science

Chronic liver inflammation is a major cause of death in patients with liver fibrosis and cirrhosis, which pose a serious health threat worldwide, and there is no effective anti-hepatic fibrosis drug. Kaempferol (KA), a flavonoid polyphenol extracted from many edible plants and traditional Chinese medicine, has been reported to possess anti-inflammatory, antioxidant, and antitumor activities and has an ameliorating effect on liver fibrosis or other fibroproliferative diseases. However, the specific regulatory mechanism of KA-reversed macrophage M1 polarization is still obscure. This study aimed to investigate the protective effects of KA on carbon tetrachloride (CCl4)-induced liver fibrosis in mice through M1 polarization. C57BL/6 mice were intraperitoneally injected with CCl4 twice weekly to induce liver fibrosis. Male mice were randomly divided into 4 groups (n= 5): the oil group, the CCl4 group, the low-dose KA-treatment CCl4 group (50 mg/kg/day KA), and the high-dose KA-treatment CCl4 group (100 mg/kg/day KA). An equal amount of solvent was given to each group by intraperitoneal injection. The results indicated that KA decreased liver pathologic changes, hepatic macrophage recruitment, and serum alanine aminotransferase levels. Notably, it reduced the activation of M1-type macrophages in the liver. The expression of proinflammatory cytokines and genes associated with M1 macrophages, such as tumor necrosis factor-α, interleukin-6, interleukin-1β, and inducible nitric oxide synthase, was also decreased. The core targets, signaling pathways, and possible mechanisms related to the M1 polarization of KA were analyzed by network pharmacology and molecular docking. Further analysis revealed that KA regulated mitogen-activated protein kinase (MAPK)/nuclear factor κB (NF-κB) signaling pathways. Finally, the results indicated that KA regulates M1 macrophage activation by modulating the MAPK/NF-κB signaling pathways. This study revealed that KA ameliorated liver injury, inflammation, and fibrosis by inhibiting macrophage M1 polarization through the MAPK/NF-κB signaling pathway, highlighting KA as a potential novel agent for the prevention and treatment of liver fibrosis. SIGNIFICANCE STATEMENT: Chronic liver inflammation is a leading cause of mortality in patients with liver fibrosis and cirrhosis, presenting a significant global health threat. Kaempferol, as a traditional Chinese medicine, effectively suppresses M1 polarization of macrophages through the mitogen-activated protein kinase/nuclear factor κB signaling pathway, thereby ameliorating liver injury, inflammation, and fibrosis. These findings underscore the potential of kaempferol as an innovative therapeutic agent for the prevention and treatment of liver fibrosis.

Lactoferrin as a therapeutic agent for attenuating hepatic stellate cell activation in thioacetamide-induced liver fibrosis

PSMP/MSMP promotes hepatic fibrosis through CCR2 and represents a novel therapeutic target

Liver fibrosis is characterized by the activation of hepatic stellate cells (HSCs) and accumulation of the extracellular matrix. There are limitations in the current therapies for liver fibrosis. Recently, oridonin was shown to induce apoptosis in HSCs. Thus, we aimed to determine the roles of oridonin in chronic liver injury and fibrosis. Liver fibrosis was induced by CCl4 in mice injected intraperitoneally with oridonin for 6 weeks. The administration of oridonin significantly attenuated liver injury and reduced ALT levels. In addition, Sirius Red staining and the expression of α‐smooth muscle actin (α‐SMA) were significantly reduced by oridonin in murine livers with fibrosis. The expression of NLRP3, caspase‐1, and IL‐1β was downregulated with the oridonin treatment. Furthermore, the expression of F4/80 in liver tissues was also decreased by oridonin treatment. These results demonstrate that oridonin ameliorates chronic liver injury and fibrosis. Mechanically, oridonin may inhibit the activity of the NLRP3 inflammasome and inflammation in the liver. These results highlight the potential of oridonin as a therapeutic agent for liver fibrosis.

The liver is a critical system for metabolism in human beings, which plays an essential role in an abundance of physiological processes and is vulnerable to endogenous or exogenous injuries. After the damage to the liver, a type of aberrant wound healing response known as liver fibrosis may happen, which can result in an excessive accumulation of extracellular matrix (ECM) and then cause cirrhosis or hepatocellular carcinoma (HCC), seriously endangering human health and causing a great economic burden. However, few effective anti-fibrotic medications are clinically available to treat liver fibrosis. The most efficient approach to liver fibrosis prevention and treatment currently is to eliminate its causes, but this approach's efficiency is too slow, or some causes cannot be fully eliminated, which causes liver fibrosis to worsen. In cases of advanced fibrosis, the only available treatment is liver transplantation. Therefore, new treatments or therapeutic agents need to be explored to stop the further development of early liver fibrosis or to reverse the fibrosis process to achieve liver fibrosis resolution. Understanding the mechanisms that lead to the development of liver fibrosis is necessary to find new therapeutic targets and drugs. The complex process of liver fibrosis is regulated by a variety of cells and cytokines, among which hepatic stellate cells (HSCs) are the essential cells, and their continued activation will lead to further progression of liver fibrosis. It has been found that inhibiting HSC activation, or inducing apoptosis, and inactivating activated hepatic stellate cells (aHSCs) can reverse fibrosis and thus achieve liver fibrosis regression. Hence, this review will concentrate on how HSCs become activated during liver fibrosis, including intercellular interactions and related signaling pathways, as well as targeting HSCs or liver fibrosis signaling pathways to achieve the resolution of liver fibrosis. Finally, new therapeutic compounds targeting liver fibrosis are summarized to provide more options for the therapy of liver fibrosis.

Arsenical poisoning may occur in industry, in three forms: (1) From inhalation of or contact with dust of inorganic compounds, (2) from inhalation of arseniuretted hydrogen, (3) from contact with organic arsenic com pounds. The inorganic compounds act as local irritants to the skin and mucosa and may have a carcinogenic effect. Arseniuretted hydrogen acts as a haemolytic agent and organic arsenic compounds as vesicants. The hepatotoxic action of inorganic arsenic compounds such as potassium arsenite, which is contained in Fowler's solution, has long been recognized by those using this solution as a therapeutic agent but has been scarcely mentioned as an industrial hazard. Skin cancer constitutes a well recognized occupational hazard. The occurrence of hydrops and ascites following the ingestion of arsenic was first recorded by Bang in 1774. Hutchinson (1895) was apparently the first to report that cirrhosis and ascites can be due to prolonged admin istration of arsenic and to draw attention to its carcino genic properties. Since then, many cases suggesting arsenic as an aetiological agent in cirrhosis of the liver have been published, ft is almost disregarded today when considering the aetiology of portal cirrhosis.

Humans and rodents with Comparative Gene Identification-58 (CGI-58) mutations manifest nonalcoholic fatty liver disease (NAFLD). Here we show that liver CGI-58 knockout (LivKO) mice fed a Western diet rapidly develop advanced NAFLD, including nonalcoholic steatohepatitis (NASH) and hepatic fibrosis. After 14 weeks of diet challenge, starting at 6 weeks of age, LivKO mice showed increased inflammatory cell infiltration and proinflammatory gene expression in the liver, which was associated with elevated plasma levels of aminotransferases. Hepatic ductular reactions, pericellular fibrosis, and bridging fibrosis were observed only in the LivKO mice. Consistently, the KO mice had a significant increase in hepatic mRNAs for fibrogenic genes. In addition, LivKO mice displayed massive accumulation of lipid droplets (LDs) in hepatocytes. LDs were also observed in the cholangiocytes of the LivKO mice, but not the floxed controls. Four of the five LD coat proteins, including perilipins 2, 3, 4, and 5, were increased in the CGI-58 KO liver. CRISPR/Cas9-mediated knockout of CGI-58 in Huh7 human hepatoma cells induced LD deposition and perilipin expression, suggesting a cell autonomous effect. Our findings establish the Western diet-fed LivKO mice as an animal model of NASH and hepatic fibrosis. These animals may facilitate preclinical screening of therapeutic agents that counter against NAFLD progression.

At present, there are no effective antifibrotic drugs for patients with chronic liver disease; hence, the development of antifibrotic therapies is urgently needed. Here, we performed an experimental and translational study to investigate the potential and underlying mechanism of quercetin treatment in liver fibrosis, mainly focusing on the impact of quercetin on macrophages activation and polarization. BALB/c mice were induced liver fibrosis by carbon tetrachloride (CCl4) for 8 weeks and concomitantly treated with quercetin (50 mg/kg) or vehicle by daily gavage. Liver inflammation, fibrosis, and hepatic stellate cells (HSCs) activation were examined. Moreover, massive macrophages accumulation, M1 macrophages and their related markers, such as tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, and monocyte chemotactic protein-1 (MCP-1) in livers were analyzed. In vitro, we used Raw 264.7 cells to examine the effect of quercetin on M1-polarized macrophages activation. Our results showed that quercetin dramatically ameliorated liver inflammation, fibrosis, and inhibited HSCs activation. These results were attributed to the reductive recruitment of macrophages (F4/80+ and CD68+) into the liver in quercetin-treated fibrotic mice confirmed by immunostaining and expression levels of marker molecules. Importantly, quercetin strongly inhibited M1 polarization and M1-related inflammatory cytokines in fibrotic livers when compared with vehicle-treated mice. In vitro, studies further revealed that quercetin efficiently inhibited macrophages activation and M1 polarization, as well as decreased the mRNA expression of M1 macrophage markers such as TNF-α, IL-1β, IL-6, and nitric oxide synthase 2. Mechanistically, the inhibition of M1 macrophages by quercetin was associated with the decreased levels of Notch1 expression on macrophages both in vivo and in vitro. Taken together, our data indicated that quercetin attenuated CCl4-induced liver inflammation and fibrosis in mice through inhibiting macrophages infiltration and modulating M1 macrophages polarization via targeting Notch1 pathway. Hence, quercetin holds promise as potential therapeutic agent for human fibrotic liver disease.

Pentoxifylline: not just for alcoholic hepatitis anymore?

Liver fibrosis is a common scarring response to chronic liver injury. It is a precursor to cirrhosis and liver carcinoma. Hepatic stimulator substance (HSS), a known liver-specific but species-nonspecific growth factor, has been shown to protect hepatocytes from various toxins. We have investigated the effects of HSS therapy on carbon tetrachloride (CCl(4))-induced and porcine-serum-mediated hepatic injury and fibrosis. We hypothesize that HSS might attenuate liver injury and fibrosis by suppressing oxidative stress, down-regulating profibrogenic factors, and blocking HSCs activation. This report demonstrated that HSS therapy diminished α-smooth muscle actin expression, decreased intrahepatic reactive oxygen species (ROS) level, and down-regulated transforming growth factor (TGF)-β1, platelet-derived growth factor (PDGF)-BB, and tissue inhibitor of metalloproteinase (TIMP)-1 expression. In addition, HSS treatment significantly protected the liver from injury by improving liver function tests and histological architecture of the liver. These results provided novel insights into the mechanisms of HSS in the protection of the liver. Our results suggested that HSS might be a therapeutic antifibrotic agent for the treatment of liver fibrosis.

Outcome of liver disease in children is mainly determined by severity and progression of liver fibrosis. Liver biopsy is the accepted standard for evaluating fibrosis but is limited by the need for sedation in children, sampling error, and risks including bleeding. The aim of the present study was to compare tools for noninvasive assessment of liver fibrosis in a paediatric cohort. Children undergoing liver biopsy for chronic liver disease were recruited and underwent transient elastography (TE). Liver biopsies were scored by a hepatohistopathologist from F0 (no fibrosis) to F4 (cirrhosis). TE was compared with biopsy score. During the study period, 104 children (62 boys) were enrolled (median age 13.6 years). Diagnosis was autoimmune liver disease in 27; nonalcoholic fatty liver disease in 37; posttransplant in 16; hepatitis B/C in 8; Wilson disease in 5; and the remainder, miscellaneous. TE was successful in all but 7 patients and was a good discriminator of significant fibrosis (≥ F2) (P < 0.001), severe fibrosis (≥ F3) (P < 0.001), and cirrhosis (F4) (P = 0.003). The area under the receiver operating characteristic curve for the prediction of ≥ F2, ≥ F3, and F4 using TE was 0.78, 0.79, and 0.96, respectively. TE performed best in children with autoimmune liver disease and in those posttransplant. The present study demonstrates that TE is a reliable tool in distinguishing different stages of liver fibrosis in paediatric patients. Thus, TE may serve as a useful adjunct to liver biopsy for diagnostic purposes providing a reliable method of noninvasively monitoring liver disease progression in children.

Objective To explore the effect factors of hepatic venous pressure gradient (HVPG) responsed to propranolol used to prevent esophageal and gastric varices bleeding (EGVB) in patients with liver cirrhosis and esophageal and gastric varices. Methods From March 2012 to September 2015, 45 patients with liver cirrhosis and esophageal and gastric varices taking propranolol after HVPG measurement were enrolled. After the second HVPG measurement, the patients were divided into HVPG response group and nonresponse group. The demographic features, clinical data, hemodynamic parameters, age, and serum propranolol concentration were compared between the two groups. The t test, chi-square test and rank sum test were performed for statistical analysis. Results Among 45 patients, 28 patients (62.2%) had response and 17 patients did not have HVPG response. There was no statistically significant difference in the age, gender, etiology for liver cirrhosis, liver function and prophylaxis indication between the two groups (all P> 0.05). The decrease of mean arterial pressure of HVPG response group and nonresponse group was 3.0 mmHg (-2.8 mmHg, 9.8 mmHg) (1 mmHg=0.133 kPa) and 1.0 mmHg (-5.5 mmHg, 9.5 mmHg), and the difference was not statistically significant (P>0.05). The basic heart rate of HVPG response group decreased from (68.6±8.6) beat per minute (bpm) before medication treatment to (59.6±7.1) bpm after medication treatment, and the difference was statistically significant (t=4.292, P<0.01). The basic heart rate of nonresponse group decreased from (68.6±7.4) bpm to (60.4±7.5) bpm, and the difference was statistically significant(t=3.217, P=0.003). The amplitudes of decrease of heart rates of response group and nonresponse group were (12.6±9.4)% and (11.5±10.3)%, respectively, and the difference was not statistically significant (t=0.358, P=0.722). After taking propranolol for 144 hours, the serum drug peak concentrations of response group and nonresponse group were 89.5 μg/L (58.5 μg/L, 158.5 μg/L) and 48.0 μg/L (33.5 μg/L, 157.5 μg/L), respectively, and there was no statistically significant difference (Z=1.371, P=0.174). Conclusions Age, gender, etiology of liver cirrhosis, liver function, prophylaxis indication, changes of pressure, decreasing of basic heart rate and serum propranolol concentration of patients with liver cirrhosis and esophageal and gastric varices were all not related with HVPG response, and could not precisely predict hemodynamics response. Precise predictiors of hemodynamic response that substitute for HVPG should be further explored. Key words: Liver cirrhosis; Hepatic venous pressure gradient; Propranolol; Hemodynamic response; Forecasting

Treatment strategies and prospects for hepatic fibrosis: Application of traditional Chinese medicine polysaccharides in modulating of the TGF-β1/Smad pathway.

BackgroundIndividuals with liver cirrhosis (LC) have an exceptionally high mortality rate. We aim to discover new biomarkers or new therapeutic targets for LC early diagnosis and treatment.MethodsThe human LC microarray datasets were obtained from the NCBI-GEO database. Firstly, differentially expressed analysis and WGCNA were performed to detect LC key genes. Protein-protein interaction (PPI) hubs revealed new regulatory mechanisms for LC-related genes. Functional enrichment and immune infiltration analyses were employed to reveal underlying mechanisms of LC progression. Then, key genes were further screened for constructing diagnostic nomogram for LC and predicting the prognosis of LC. CB-Dock-2 was employed to explore potential therapeutic agents for LC based on the identified key genes.Finally, immunohistochemistry was performed to detect the expression levels of the key genes in liver tissues from patients with liver cirrhosis and healthy controls.ResultsThe integrated LC dataset identified 749 LC key genes by intersecting WGCNA and differential expression analyses. PPI analysis identified 15 genes as key regulators in LC development, predominantly enriched in inflammatory and immune regulatory pathways. We conducted an in-depth analysis of two leukocyte transendothelial migration (LTM)-associated genes (MMP2 and ITGB2) among these 15 hub genes, revealing their involvement in the construction of the immune microenvironment in LC. A nomogram based on MMP2 and ITGB2 had favorable diagnostic performance for LC, and ITGB2 showed potential in predicting LC prognosis. Through molecular docking approaches, this study further identified several antioxidant and anti-inflammatory agents with therapeutic potential for LC. Immunohistochemical staining revealed significantly elevated protein levels of MMP2 and ITGB2 in liver tissues from cirrhotic patients compared to healthy controls.ConclusionsLTM-associated genes, namely MMP2 and ITGB2, are associated with LC and may represent candidate biomarkers or potential targets requiring further experimental validation.