Hepatoprotective Components Research Articles

Effects of compound Anoectochilus roxburghii (Wall.) Lindl. oral liquid on relative metabolic enzymes and various biochemical indices in Wistar rats with isoniazid-induced liver injury

Isoniazid (INH) is the first-line anti-tuberculosis drug in clinical practice, and its main adverse effect is drug-induced liver injury (DILI). This study aimed to investigate the hepatoprotective effect of Compound Anoectochilus roxburghii (Wall.) Lindl. Oral Liquid (CAROL) and to provide a new strategy for the search of potential drugs against INH-induced liver injury in Wistar rats. Animal experiment was based on INH (100 mg/kg) induced liver injury to explore the intervention effects of CAROL at doses of 1.35, 2.70, and 5.40 mL/kg. LC-QTOF-MS/MS was used to identify hepatoprotective components in CAROL and its’ exposed components in rat serum. The hepatoprotective effect of CAROL was evaluated by pathological observation of rat liver tissue and changes in levels of biochemical indices and cytokines in serum or liver tissue. Of the 58 hepatoprotective components identified, 15 were detected in the serum of rats with liver-injured treated by high-dose CAROL. Results of animal experiments showed that the levels of various biochemical indexes and cytokines were significantly reversed with CAROL intervention. In particular, the expression level of cytokeratin-18 and high-mobility group box 1, as specific and sensitive indicators of DILI, was significantly reduced in the serum of rats with CAROL intervention compared with the INH model group. The same reversal was observed in the levels of TBIL, ALP, ALT, and AST in serum, as well as in the levels of TNF-α, IL-6, SOD, and MDA in liver tissue. For INH-metabolizing enzymes, an evident expression inhibition was observed in N-acetyltransferase 2 and glutathione S-transferases with CAROL intervention, which may be the key to controlling INH hepatotoxicity. CAROL has a favorable hepatoprotective effect on INH-induced liver injury. This study takes the first step in studying the hepatoprotective mechanism of CAROL against INH hepatotoxicity and provides reference for wider clinical applications.

Total cucurbitacins from Herpetospermum pedunculosum pericarp do better than Hu-lu-su-pian (HLSP) in its safety and hepatoprotective efficacy.

The pericarp of Herpetospermum pedunculosum (HPP) has traditionally been used for treating jaundice and hepatitis. However, the specific hepatoprotective components and their safety/efficacy profiles remain unclear. This study aimed to characterize the total cucurbitacins (TCs) extracted from HPP and evaluate their hepatoprotective potential. As a reference, Hu-lu-su-pian (HLSP), a known hepatoprotective drug containing cucurbitacins, was used for comparison of chemical composition, effects, and safety. Molecular networking based on UHPLC-MS/MS identified cucurbitacin B, isocucurbitacin B, and cucurbitacin E as the major components in TCs, comprising 70.3%, 26.1%, and 3.6% as determined by RP-HPLC, respectively. TCs treatment significantly reversed CCl4-induced metabolic changes associated with liver damage in a dose-dependent manner, impacting pathways including energy metabolism, oxidative stress and phenylalanine metabolism, and showed superior efficacy to HLSP. Safety evaluation also showed that TCs were safe, with higher LD50 and no observable adverse effect level (NOAEL) values than HLSP. The median lethal dose (LD50) and NOAEL values of TCs were 36.21 and 15mg/kg body weight (BW), respectively, while the LD50 of HLSP was 14mg/kg BW. In summary, TCs extracted from HPP demonstrated promising potential as a natural hepatoprotective agent, warranting further investigation into synergistic effects of individual cucurbitacin components.

Systematic screening of hepatoprotective components from traditional Chinese medicine: Zuojin Pill as an example

Ethnopharmacological relevanceZuojin Pill (ZJP), composed of Coptis chinensis Franch. and Euodia ruticarpa (A. Juss.) Benth. in a mass ratio of 6:1, is a famous traditional Chinese medicine (TCM) formula recorded in “Danxi's Experiential Therapy”, an ancient medical book from the Ming Dynasty of China. It is used to treat liver fire invading the stomach, which is caused by liver stagnation transforming into fire and disharmony between the liver and stomach. Aim of the studyTo develop a systematic strategy to screen hepatoprotective components from TCM using ZJP as a model sample. Materials and methodsA CCl4-induced mouse model of acute liver injury was used for the verification of the hepatoprotective effects of ZJP. UPLC-Q-Exactive Plus Orbitrap MS/MS was used for the identification of the components in mouse serum after intragastric administration of ZJP. The hepatoprotective activities of the components found in mouse serum were tested in primary cultured mouse hepatocytes induced by CCl4. ResultsNine components with significant hepatoprotective activity including berberine, epiberberine, coptisine, palmatine, jatrorrhizine, rutaecarpin, dehydroevodiamine, evocarpine and chlorogenic acid were successfully screened out. ConclusionsOur developed strategy has the advantages of high efficiency and low cost, and would provide a powerful tool for screening potential hepatoprotective components from TCM.

Identification and validation of targets of swertiamarin on idiopathic pulmonary fibrosis through bioinformatics and molecular docking-based approach

BackgroundSwertiamarin is the main hepatoprotective component of Swertiapatens and has anti-inflammatory and antioxidation effects. Our previous study showed that it was a potent inhibitor of idiopathic pulmonary fibrosis (IPF) and can regulate the expressions of α-smooth muscle actin (α-SMA) and epithelial cadherin (E-cadherin), two markers of the TGF-β/Smad (transforming growth factor beta/suppressor of mothers against decapentaplegic family) signaling pathway. But its targets still need to be investigated. The main purpose of this study is to identify the targets of swertiamarin.MethodsGEO2R was used to analyze the differentially expressed genes (DEGs) of GSE10667, GSE110147, and GSE71351 datasets from the Gene Expression Omnibus (GEO) database. The DEGs were then enriched with Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis for their biological functions and annotated terms. The protein-protein interaction (PPI) network was constructed to identify hub genes. The identified hub genes were predicted for their bindings to swertiamarin by molecular docking (MD) and validated by experiments.Results76 upregulated and 27 downregulated DEGs were screened out. The DEGs were enriched in the biological function of cellular component (CC) and 7 cancer-related signaling pathways. Three hub genes, i.e., LOX (lysyl oxidase), COL5A2 (collagen type V alpha 2 chain), and CTGF (connective tissue growth factor) were selected, virtually tested for the interactions with swertiamarin by MD, and validated by in vitro experiments.ConclusionLOX, COL5A2, and CTGF were identified as the targets of swertiamarin on IPF.

Multi-target-based polypharmacology prediction (mTPP): An approach using virtual screening and machine learning for multi-target drug discovery

Polypharmacology has become a new paradigm in drug discovery and plays an increasingly vital role in discovering multi-target drugs. In this context, multi-target drugs are a promising approach to treating polygenic diseases. Many in-silico prediction methods have been developed to screen active molecules acting on multiple targets. The relationship between the action of multiple targets and the drug's overall efficacy is significant for developing multi-target drugs. So, the prediction method for this relationship urgently needs to be developed. This paper introduces multi-target-based polypharmacology prediction (mTPP), an approach using virtual screening and machine learning to explore the relationship. To predict the activity of the potential hepatoprotective components, the data on the binding strength of a single ingredient with multiple targets and the proliferation rate of the compounds against acetaminophen (APAP)-induced injury L02 cells were all used to construct the mTPP model by Multi-layer Perceptron (MLP), Support Vactor Regression (SVR), Decision Tree Regressor (DTR), and Gradient Boost Regression (GBR) algorithms. Compared with MLP, SVR, and DTR algorithms, GBR algorithms showed the best performance with R2test = 0.73 and EVtest = 0.75. In addition, 20 candidates with potential effects against drug-induced liver injury (DILI) were predicted by the mTPP model. Furthermore, 2 of the 20 candidates, Chelerythrine and Biochanin A, were applied to evaluate the model's accuracy. The results showed that Chelerythrine and Biochanin A could improve the viability of APAP-induced injury cells. Thus, the mTPP model is hoped to help develop polypharmacology and discover multi-target drugs.

Discovery of possible hepatoprotective components from lychee pulp phenolic extract by online knockout methods

Lychee pulp phenolic extract (LPPE) possessed protective effects against nonalcoholic fatty liver disease (NAFLD), while the specific components concerned were unclear. In this study, twelve major components in LPPE were identified by LC-MS and prepared by on line knockout methods. Two antioxidant methods companied with hepatocyte steatosis assay were used to evaluate the contribution of twelve components to the hepatoprotective effects of LPPE. The identified components include four procyanidins and eight flavonoids, they all play active roles in the oxygen radical absorbance capacity of LPPE, while only B-type procyanidin trimer and quercetin 3-O-rutinoside-7-O-rhamnoside exert positive effects on cellular antioxidant activity of LPPE. Six components including procyanidin B2, epicatechin, two quercetin glycosides, and two procyanidin trimers are considered as contributors to lipid accumulation inhibitory effects of LPPE with B-type procyanidin trimer being identified as the main contributor. All the twelve components were recognized as hepatoprotective components against NAFLD in LPPE.

Schisantherin D from Schisandra chinensis (Turcz.) Baill. exhibits anti-liver fibrosis capacity via modulating ETBR involved signaling, an in vitro and in vivo study

Excess levels of chemical hepatotoxicants (alcohol, aflatoxin B1), oxidative drugs (acetaminophen) and some cytokines (ET-1, TGF-β1) can induce chronic or acute liver injury. After these, the severe hepatic disease, especially the liver fibrosis (LF) occurs without taking measures, which brings threat to human health. The dibenzocyclooctadiene lignans of S. chinensis (SCDLs) were found to act as the hepatoprotective components via blocking endothelin B receptor (ETBR). While study on its anti-LF mechanisms especially for its refined compound of schisantherin D (SC-D) is still a lack. So this study aims to investigate the anti-fibrosis effect of SC-D with in vitro and in vivo assays. Bioinformatics analysis revealed the close relations of ETBR to Smad2, Smad3, Nrf2, etc. in LF-related signaling pathways (such as TGF-β/Smad and Nrf2/ARE). Histopathological staining on livers showed the recovery trend in SC-D treated LF mice. SC-D also modulated expressions of ETBR and fibrosis or anti-oxidative related proteins (such as TIMP1, p-Smad2/3, Nrf2, Smad7, etc.) in LF mice livers. Serum levels of TNF-α, COLI, ALT, AST and LDH in SC-D treated mice were also downregulated compared with LF mice, and upregulated expression of GSH. In vitro studies, SC-D also modulated expressions of LF-related proteins to the normal tendency in LX-2 cell, while weakened its anti- LX-2 proliferation effect by transfections of si-Smad7 or si-Nrf2. Accordingly the anti-LF approach of SC-D showed relations with modulating ETBR linked fibrosis and anti-oxidative related signaling. Also, Smad7 and Nrf2 might be the key factors for SC-D mediated anti-LF effect.

Spectrum-Effect Relationships Between High-Performance Liquid Chromatography Fingerprints and Hepatoprotective Activities of Cuscutae Semen.

Cuscutae Semen (CS) is a commonly used hepatoprotective traditional Chinese medicine, but the chemical components responsible for its hepatoprotective activity are unclear. The purpose of this study was to evaluate the spectrum-effect relationships between HPLC fingerprints and hepatoprotective effects of CS, and to identify its bioactive components. Phytochemical isolation of CS extracts was first carried out and 14 potential bioactive compounds were obtained. Chemical fingerprinting was performed on 27 batches of CS from different sources by HPLC, and further analyzed by similarity analysis (SA) and hierarchical clustering analysis (HCA). Pharmacodynamic testing was performed in a CCl4-induced, acute liver injury cell model to assess the hepatoprotective activity of CS by measuring the cell viability and levels of alanine transaminase (ALT) and aspartate aminotransferase (AST). Bivariate correlations analysis (BCA) and orthogonal projections to latent structures (OPLS) were used to analyze the spectrum-effect relationships of CS. The results showed that the chemical fingerprints of CS were closely correlated with its hepatoprotective activity. Peaks 1, 10, 18, 19, 21, 22, and 24 might be potential hepatoprotective compounds in CS, and the validation experiments of isolated compounds indicated that chlorogenic acid (P10), hyperoside (P21), isoquercitrin (P22), and astragalin (P24) were the main hepatoprotective components. By combining chemical fingerprints with hepatoprotective evaluation, the present study provides important guidance for QC and clinical use of CS. (1) Ten potential bioactive compounds were isolated from CS; (2) The spectrum-effect relationship of CS was molded by HPLC and analysed by OPLS and BCA. (3) Four compounds including chlorogenic acid were the main hepatoprotective components.

UPLC-Q-Exactive-MS analysis for hepatotoxicity components of Evodiae Fructus based on spectrum-toxicity relationship

Evodiae Fructus (EF) is generally divided into three categories: small flower EF (SEF), medium flower EF (MEF) and big flower EF (BEF) in commodity circulation according to the size of the fruit. It is a well-known and frequently used herbal medicine for treating gastrointestinal disorder-related stomachache and headache, which has aroused wide attention for its hepatotoxicity. However, reports about hepatotoxicity is controversial and hepatotoxic components are inconclusive. The study aimed to explain the controversial hepatotoxicity of EF and screen the components associated with hepatotoxicity of EF based on the spectrum-toxicity relationship. UPLC fingerprints of 39 batches of EF collected from different regions were established. Combined with the results of L02 cell viability assays, the spectrum-toxicity relationship was investigated on the basic of orthogonal partial least squares (OPLS). The results of the research demonstrated that the toxicity of EF was obviously various among the different categories, in particularly, SEF was with less toxicity, MEF except for adulterants and BEF had mild toxicity and adulterants of MEF (A-MEF) produced more damage to L02 cell and no regions specificity in hepatotoxicity of EF. Thereinto, samples, the contents of which do not meet the requirements of Chinese Pharmacopoeia, were adulterants. It was worth noting that P11, P17, P20 and P25 were closely related to hepatotoxicity of EF and they were respectively identified as limonin (LIM), evodiamine (EVO), 1-methyl-2-nonyl-4(1H)-quinolone (MNQ), and 1-methyl-2-undecyl-4(1H)-quinolone (MUQ) by UPLC-Q-Exactive-MS. The hepatoprotection of P11 and hepatotoxicity of P17 were consistent with the results of spectrum-toxicity relationship. In summary, A-MEF was more toxic than other categories and SEF was less toxic than the others. It was noteworthy that EVO was the main hepatotoxic component of EF and LIM was the main hepatoprotective component of EF. The results provided worthy evidence for better utilization and development of EF.

Hepatoprotective effects of extracts, fractions and compounds from the stem bark of Pentaclethra macrophylla Benth: Evidence from in vitro and in vivo studies

AimTo identify the bioactive hepatoprotective components of the ethanol extract of Pentaclethra macrophylla stem bark using in vitro and in vivo approaches. MethodsThe bioguided-fractionation of the ethanol extract was based on the substances’ capacity to prevent in vitro, the lipid peroxidation of hepatocytes’ membranes induced by hydrogen peroxide. For the in vivo hepatoprotective test, mice were treated orally with the ethyl acetate (EtOAc) fraction of the ethanol extract at doses of 50 and 75 mg/kg/day for one week and subjected to d-galactosamine/lipopolysaccharide (GaIN/LPS)-induced hepatotoxicity. Blood samples were collected for alanine aminotransferase (ALAT), aspartate aminotransferase (ASAT), TNF-α and IL-1β assays. The liver was harvested for histological and biochemical (proteins, glutathione (GSH), catalase and superoxide dismutase (SOD)) analysis. ResultsThe ethanol extract and fractions induced concentration-dependent inhibition of lipid peroxidation (IC50: 3.21–48.90 μg/mL) greater than that of silymarin (IC50: 117.4 μg/mL). The purification of the sub-fractions of EtOAc fraction yielded: (7R)-7-hydroxyhexacosanoic acid (1), (7R)-1-(7-hydroxyhexacosanoyl) glycerol (2), bergenin (3), 11-O-galloylbergenin (4), 2-hydroxymethyl-5-(2-hydroxypropan-2-yl)phenol (5), β-sitosterol 3-O-β-d-glucopyranosyl (6) and β-sitosterol (7)), among which 11-O-galloylbergenin (IC50:1.8 μg/mL) was the most effective. The EtOAc fraction significantly reduced the serum level of ALAT, ASAT and TNF-α in vivo. This EtOAc fraction increased the liver protein content and protected the liver against structural damages, but did not boost the endogenous antioxidant parameters. ConclusionThe stem bark of Pentaclethra macrophylla possesses hepatoprotective effects that may result from its capacity to inhibit lipid peroxidation and could be attributed to its active components 3, 4 and 2.

Total Flavonoids of Crocus sativus Petals Release tert-Butyl Hydroperoxide-Induced Oxidative Stress in BRL-3A Cells.

Antioxidant and hepatoprotective activities in vitro of saffron petals were examined in this study for better utilizing saffron (Crocus sativus L.) biowaste. Using the DPPH and ABTS radical scavenging method, we compared the antioxidant activity and the content of total flavonoid extracts from petals (TFESP), stamens (TFESS), and both saffron petals and stamens (TFEMS). The results showed that the antioxidant capacity and the flavonoid content of TFESP were higher than those of TFESS and TFEMS. Then, the hepatoprotective activity of TFESP was determined, and the silymarin was used as a positive control. The main components of TFESP were analysed by ultrahigh performance liquid chromatography (UPLC) photodiode array (PDA)/mass spectrometry (MS) and nuclear magnetic resonance (NMR). The result showed that (1) TFESP could release oxidative liver injury induced by tert-butyl hydroperoxide (t-BHP). (2) TFESP could reduce the accumulation of reactive oxygen species (ROS); enhance the activity of superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH); and then improve the total antioxidant capacity (T-AOC) in BRL-3A cells. (3) TFESP could enhance the expression of B-cell lymphoma-2 (BCL-2) and decrease the expression of caspase-3 and caspase-9; increase the expression of Kelch-like ECH-associated protein-1 (Keap-1), nuclear factor, erythroid 2-related factor 2 (Nrf2), superoxide dismutase, and heme oxygenase 1 (HO-1); and downregulate inducible nitric oxide synthase (INOS), interleukin-6 (IL-6), and nuclear factor kappa B-9 (NF-κB-9). (4) The main hepatoprotective component of TFESP was identified as kaempferol-3-o-sophoroside. The mechanism may be that kaempferol-3-o-sophoroside can protect t-BHP-induced cell injury by regulating the expression of antioxidant, antiapoptotic, and anti-inflammatory genes. Thus, saffron petals are a potential hepatoprotective resource worthy of development.

Antibiotic prophylaxis of purulent and septical complications during surgical treatment of complicated gallstone disease on background of liver cirrhosis

Purpose of the study. The goal of this investigation was to determine the main risk factors in development of purulent and septical complications during surgical treatment of complicated gallstone disease on background of liver cirrhosis.
 Materials and methods. We retrospectively reviewed medical records of 247 patients who undergone treatment in minimally invasive surgery centre of Zhytomyr regional clinical hospital during 2009–2018. All patients with complicated gallstone disease were divided in two groups: group 1 – patients with concomitant liver cirrhosis – 79(31,98%), and group 2 – patients without liver cirrhosis – 168 (68,02%). Theinclusion criteria were presents of complicated gallstone disease (acute calculous cholecystitis, choledocholithiasis with obstructive jaundice and Mirizzi syndrome), and verified liver cirrhosis. Patients with oncological history, immunodeficiency and morbid obesity were excluded. Liver cirrhosis was staged by Child-ТurcotteРugh system. In all cases patient’s condition was assessed by APACHE II scoring system and patients with cirrhotic lesion were additionally analyzed by MELD score. Acute calculous cholecystitis was diagnosed in 185 patients: group 1 – 68(Child A – 36, Child B – 31, Child C – 1), group 2 – 117 patients. The signs of cholodecholithiasis with obstructive jaundice were present in 49 cases: group 1 – 7 (Child A – 5, Child B – 1, Child C – 1), group 2 – 42. Mirizzi syndrome was verified in 9 cases: group 1 – 2 (Child A – 1, Child B – 1, group 2 – 7.
 Results and discussion. In all patients treatment was started in conservative way that included detoxic, antibacterial and hepatoprotective components. In group of control early operative tactic in cases with acute calculous cholecystitis was preferred. Antimicrobial prophylaxis was performed in cases of severe and moderate calculous cholecystitits with use of cephalosporines of 2 generation in moderate case, and protected cephalosporines of 3 generation in combination with metronidazol in severe. In main group providing of antimicrobial therapy was performed very carefully, because of higher risk of hepato-renal insufficiency. The early de-escalation therapy was mandatory performed. Purulent complications occurred in 13,2% of patient with liver cirrhosis in comparison with control group with 1,7% of complications.
 Conclusion. The treatment of complicated gallstone disease in patients with liver cirrhosis is very risky in case of postoperative purulent complications. In case of Child A stage of cirrhosis the treatment is safe, and the incidents of purulent complications is the same like in the absence of cirrhosis. Administration of antibiotics in cirrhotic should be very careful because of higher risk of hepato-renal insufficiency. The early de-escalation therapy should be mandatory performed. The «gold» standard of empirical antimicrobial therapy is the use of cephalosporines of 2 and 3 generation.

Metabolomics Reveals the Efficacy of Caspase Inhibition for Saikosaponin D-Induced Hepatotoxicity.

Saikosaponin d (SSd) is a major hepatoprotective component of saikosaponins derived from Radix Bupleuri, which was also linked to hepatotoxicity. Previous studies have demonstrated that caspases play a key role in SSd-induced liver cell death. Our in vitro and in vivo studies also showed that treatment with caspase inhibitor z-VAD-fmk could significantly reduce the L02 hepatocyte cells death and lessen the degree of liver damage in mice caused by SSd. In order to further reveal the underlying mechanisms of caspase inhibition in SSd-induced hepatotoxicity, mass spectrometry based untargeted metabolomics was conducted. Significant alterations in metabolic profiling were observed in SSd-treated group, which could be restored by caspase inhibition. Bile acids and phospholipids were screened out to be most significant by spearman correlation analysis, heatmap analysis and S-Plot analysis. These findings were further confirmed by absolute quantitation of bile acids via targeted metabolomics approach. Furthermore, cytokine profiles were analyzed to identify potential associations between inflammation and metabolites. The study could provide deeper insight into the hepatotoxicity of SSd and the efficacy of caspase inhibition.

Selective recognition and preliminary separation of hepatoprotective component silybin from milk thistle seeds by the prepared core–shell magnetic molecularly imprinted polymer

Using hepatoprotective component silybin as the template, MAA as the functional monomer, EGDMA as the cross linker, acetonitrile as the solvent, and Fe3O4 as the magnetic substrate, the silybin magnetic molecularly imprinted polymer(S-MIP) was successfully prepared by the reverse atom transfer radical precipitation polymerization(RATRP), and non-ion imprinted polymer(NIP) was also synthesized by the same procedure only in the absence of silybin. On the basis of computer simulation and L9(34) orthogonal array design matrix, the optimal preparation conditions were finally obtained as follows that the molar ratio of silybin, MAA, and EGDMA was 1:5:60. The prepared materials were respectively characterized by SEM, XRD, FT-IR, VSM and TGA, and the kinetic analyses explored that the adsorption process of S-MIP was better describled by the pseudo-second order kinetic equation, meanwhile, scatchard analysis illustrated that the heterogeneous binding sites existed in S-MIP. Both the selectivity factor α (2.232) and the relative selectivity factor β (2.201) of S-MIP showed the aim material had the excellent specific recognition capacity and selection adsorption property to silybin. Additionally, the recycling reuse experiments explored S-MIP was of a good stability and could be used repeatedly at least five times with only 15.8% decrease of adsorption capacity when applied in the fifth experiment, and the evaluation experiments by HPLC for silybin extraction from the medicinal herb milk thistle seeds indicated that S-MIP had the potential application in separating the natural hepatoprotective component silybin from the herbal plants.

Toralactone glycoside in Cassia obtusifolia mediates hepatoprotection via an Nrf2-dependent anti-oxidative mechanism

Cassia obtusifolia L. (Leguminosae) seeds are a well-known medicinal food in East Asia and are used to clear liver heat, sharpen vision, lubricate the intestines, and promote bowel movement. The aims of the present study were to identify the hepatoprotective components of C. obtusifolia seeds by bioactivity-guided isolation and to elucidate their mechanisms of action. Ten phenolic glycosides were isolated from the most active ethyl acetate fraction, and their chemical structures were elucidated by spectroscopic analyses. Among the isolated compounds, toralactone 9-O-gentiobioside (5) had the highest hepatoprotective efficacy against tert-butylhydroperoxide-induced cell death in HepG2 cells. Immunoblotting and real-time polymerase chain reaction analyses revealed that the hepatoprotective effects were exerted through nuclear factor erythroid-2-related factor 2 (Nrf2)-dependent antioxidative signaling. Together, these results provide insights into the effects of this medicinal plant as well as a basis for developing hepatoprotective agents as pharmaceuticals and/or nutraceuticals.

Protective role and antioxidant activity of arabic gum againsttrichloroacetate-induced toxicity in liver of male rats

Investigation of functional hepatoprotective components from plants origin is a new model for drug evolution. The study examined the protective effects of Arabic gum (AG) induction on liver serum markers, antioxidant enzymes and lipid peroxidation in liver tissue against trichloroacetate (TCA) induced hepatotoxicity in male albino rats. Also, the chemical constituents and antioxidant assay of aqueous extract of AG was determined. TCA at the dose of 50 mg/kg for 2 months produced hepatotoxicity as examined by the significant increase of serum activities of ALT, AST, ALP and conjugated bilirubin level in the TCA treated animals alone and decrease in total protein and albumin levels. Pretreatment with AG aqueous extract (0.5 g/kg/day) significantly (P< 0.05) lowered the serum enzyme activities and increased in total proteins and albumin level. The antioxidant studies showed that the activities of hepatic SOD, CAT and GPx were decreased in TCA induced animals with significantly elevated in MDA content which is reversed in pretreatment with aqueous extract of AG. Histopathology of the liver sections confirmed that the AG extract ameliorated hepatic damage induced by TCA. In conclusion, the study demonstrated that AG supplementation for 2 months in TCA induced toxicity in rats benefited hepatic antioxidant status and improved liver injury and damage in male albino rats exposed to TCA.

Heat shock proteins 27 and 70 contribute to the protection of Schisandrin B against d-galactosamine-induced liver injury in mice.

Schisandrin B is a hepatoprotective component isolated from a traditional Chinese herb, Schisandra chinensis (Turcz.) Baill. This study determined the effect of Schisandrin B on d-galactosamine -induced liver injury and the role of heat shock proteins 27 and 70 against liver injury in mice. Acute liver injury was induced by intraperitoneal injection of d-galactosamine to mice, and Schisandrin B was given orally. The protein and gene expression of heat shock proteins 27 and 70 were detected by western blot and real-time quantitative polymerase chain reaction, respectively. Liver tissues were subjected to histological evaluation, and the activities of alanine aminotransferase and aspartate aminotransferase in the serum were measured. Pretreatment of Schisandrin B significantly attenuated d-galactosamine-induced liver injury in mice. This result was evidenced by improved alteration of histopathological hepatic necrosis and reduced alanine aminotransferase and aspartate aminotransferase activities in the serum. The hepatoprotective effect was accompanied with overexpression of heat shock proteins 27 and 70 both at the protein and mRNA levels. However, the aforementioned actions of Schisandrin B were all markedly suppressed by the heat shock protein inhibitor quercetin. Heat shock proteins 27 and 70 were involved in the protective effect of Schisandrin B against d-galactosamine-induced liver injury in mice.

Hepatoprotective potential of antioxidant potent fraction from Urtica dioica Linn. (whole plant) in CCl4 challenged rats

The aim of the present study was to isolate hepatoprotective component from Urtica dioica Linn. (whole plant) against CCl4-induced hepatotoxicity in-vitro (HepG2 cells) and in-vivo (rats) model. Antioxidant activity of hydro alcoholic extract and its fractions petroleum ether fraction (PEF), ethyl acetate fraction (EAF), n-butanol fraction (NBF) and aqueous fraction (AF) were determined by DPPH and NO radicals scavenging assay. Fractions were subjected to in-vitro HepG2 cell line study. Further, the most potent fraction (EAF) was subjected to in-vivo hepatoprotective potential against CCl4 challenged rats. The in-vivo hepatoprotective active fraction was chromatographed on silica column to isolate the bioactive constituent(s). Structure elucidation was done by using various spectrophotometric techniques like UV, IR, 1H NMR, 13C NMR and MS spectroscopy. Ethyl acetate fraction (EAF) of hydro-alcoholic extract of U. dioica possessed the potent antioxidant activity viz. DPPH (IC50 78.99±0.17μg/ml) and NO (IC50101.39±0.30μg/ml). The in-vitro HepG2 cell line study showed that the EAF prevented the cell damage. The EAF significantly attenuated the increased liver enzymes activities in serum and oxidative parameters in tissue of CCl4-induced rats, suggesting hepatoprotective and anti-oxidant action respectively. Column chromatography of most potent antioxidant fraction (EAF) lead to the isolation of 4-hydroxy-3-methoxy cinnamic acid (ferulic acid) which is responsible for its hepatoprotective potential. Hence, the present study suggests that EAF of hydro-alcoholic extract has significant antioxidant and hepatoprotective potential on CCl4 induced hepatotoxicity in-vitro and in-vivo.

Methodology to nanoencapsulate hepatoprotective components from Picrorhiza kurroa as food supplement

Abstract Picrorhiza kurroa root and rhizome powder of extract is well-recognized neutraceutical for healthy liver functioning. The major hepatoprotective components (picrosides I and II) of P. kurroa extract shows poor intestinal absorption and poor bioavailability due to sparingly aqueous solubility. To improve bioavailability of these biomolecules, a nanoformulation of this extract has been developed with pluronic-F-68 copolymer based biodegradable PLA nanoparticles by nanoprecipitation method. This method showed high encapsulation efficiency as 60.1±2.8% (for picroside I) and 67.2±7.4% (for picroside II). The hydrodynamic (by zeta size) and actual size (by TEM) revealed homogenously rounded nanoparticles of size 174±34 nm and 154±12 nm respectively. This formulation showed an initial burst and half release up to 27±3 h, followed by a sustained release (up to 210±10 h). Nanoformulation of this traditional herbal extract provides neutraceutical with value addition property for better hepato-protection by enhancing intestinal absorption and bioavailability.

Adenosine, a hepato-protective component in active hexose correlated compound: Its identification and iNOS suppression mechanism

Supplementation of active hexose correlated compound (AHCC) improved the prognosis of postoperative hepatocellular carcinoma patients. Excess production of nitric oxide (NO) by inducible NO synthase (iNOS) is an inflammatory biomarker in liver injury. AHCC suppressed iNOS induction in hepatocytes, suggesting that AHCC has a potential liver-protective effect. However, the active component in AHCC responsible for NO suppressive activities has not been identified. The objective of this study was to identify this NO suppressive component and to investigate its mechanisms of action. AHCC was subjected to fractionation by cation exchanger, size exclusion chromatography, and normal- and reversed-phase HPLC. Aliquots of the fractions were added to primary cultured rat hepatocytes stimulated with interleukin (IL)-1β, and NO production was assayed. By activity-guided fractionation and electron spray ionization mass spectrometry analysis, adenosine was identified as one of the NO suppressive components in AHCC. Adenosine inhibited NO production, and reduced the expression of iNOS protein and mRNA. It had no effects on IκB degradation, but it inhibited NF-κB activation. Adenosine also inhibited the upregulation of type I IL-1 receptor (IL-1RI). Experiments with iNOS promoter-luciferase constructs revealed that adenosine decreased the levels of iNOS mRNA at the promoter transactivation and mRNA stabilization steps. Adenosine decreased the expression of the iNOS gene antisense transcript, which is involved in iNOS mRNA stability. Adenosine in AHCC suppressed iNOS induction by blocking NF-κB activation and the upregulation of the IL-1RI pathways, resulting in the inhibition of NO production.