ABSTRACT
An immunologically stressed rat model was used in a metabolomics study on the ability of Paeoniae Rubra Radix to reduce the liver toxicity of Psoraleae Fructus. Different groups of rats were given the extracts of Psoraleae Fructus and Psoraleae Fructus together with Paeoniae Rubra Radix or combined with a non-toxic dose of lipopolysaccharide (LPS). The biochemical indices of liver function and pathological changes in liver tissue were used to evaluate histopathological changes. UHPLC-QTOF/MS was used to analyze the metabolic profile of serum samples, combined with principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) methods. The HMDB database and Metabo Analyst online tool were used for biomarker identification and metabolic pathway-enrichment analysis. The results show that the co-treatment Psoraleae Fructus and LPS resulted in significant liver injury, indicated by the elevation of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities, as well as obvious pathological changes. Liver injury was significantly decreased by treatment with Paeoniae Rubra Radix. Metabolomic analysis showed that the addition of Paeoniae Rubra Radix ameliorated the abnormal serum metabolism in rats mainly through regulation of arachidonic acid metabolism and glycerophospholipid metabolism pathways.
ABSTRACT
Using the idiosyncratic lipopolysaccharide (LPS)-mediated hepatotoxicity model as a positive control, liver injury induced by Cortex Dictamni aqueous extract (AE) or Cortex Dictamni ethanol extracts (EE) was evaluated. Idiosyncratic hepatotoxicity model was established in rats [Institutional Animal Care and Use Committee (IACUC)-2018-008] by injecting LPS at a dosage of 2.8 mg·kg-1. Rats were randomly divided into 10 groups. The plasma levels of liver function biomarkers such as alanine transaminase (ALT), aspartate aminotransferase (AST) were measured. Histological changes (HE staining), hepatocellular apoptosis and the content of cytokines of liver were measured. Network pharmacology was used to analyze the relationship between chemical components and immunity in Cortex Dictamni. Compared with the control group, the doses (25, 50 g·kg-1) of AE or EE had no significant changes in ALT, AST and liver pathology (P>0.05). The doses of 4.2 g·kg-1 of AE or EE+LPS groups exhibited an elevation in ALT, AST and serum cytokines (P<0.01). Disorder of liver lobular arrangement and irregular island-like or massive necrosis of liver cells were observed in these groups. Network pharmacology shows that Cortex Dictamni may directly or indirectly participate in the process of immunomodulation. We found that Cortex Dictamni regulated 15 core targets and affected 19 pathways, including apoptosis, TNF-α, NF-kappa B signaling pathways. These results suggest that Cortex Dictamni can induce idiosyncratic hepatotoxicity and the water extract can induce more serious liver injury then ethanol extract of Cortex Dictamni. These findings provide a reference for elucidating the idiosyncratic hepatotoxicity induced by Cortex Dictamni.
ABSTRACT
In this study, we used a mathematic-based modeling system to screen the cytokines that are sensitive to Zhuangguguanjie wan (ZGW)-induced idiosyncratic liver injury. The values of 27 cytokines were used as the data source in rat liver of lipopolysaccharide (LPS) + ZGW group. The alanine aminotransferase (ALT) activity value of liver function indexes was used as the outcome evaluation index of liver injury. Cytokines of ZGW-induced idiosyncratic liver injury were screened using Logistic regression, random forest method, LASSO Logistics regression and method of combining rule discovery algorithm with LASSO, and cytokines filtered out were revalued in THP1 macrophage. Susceptible cytokine combinations:interleukin-1β (IL-1β), epidermal growth factor (EGF) and interleukin-18 (IL-18) closely related to ZGW-induced idiosyncratic liver injury were obtained after preliminary screening analysis. The result of revalued in THP1 showed that the ethanolic extract of ZGW (EtZ) combined with IL-1β or IL-18 synergistically enhanced tumor necrosis factor-α (TNF-α) secretion in THP1 macrophage, and EtZ combined with IL-1β significantly enhanced interleukin-6 (IL-6) secretion in THP1 macrophage, but EtZ combined with EGF markedly inhibited IL-6 secretion in THP1 macrophage. The results suggest that the sensitive cytokines that can be characterized in the ZGW-induced idiosyncratic liver injury are IL-1β and IL-18, which provides a basis for screening the ZGW-induced idiosyncratic liver injury patients, and a new experimental evidence for clinical safety medication and risk prevention of ZGW.
ABSTRACT
Objective: To analyze the correlation of idiosyncratic hepatotoxicity of Zhuangguguanjie Wan (ZGW) with 27 cytokines. Methods: After 12 h fasting, SD rats were ig with ZGW at a single dose of 3.8 g/kg, and injected with lipopolysaccharide (LPS) at a dosage of 2.8 mg/kg via tail vein after 2 h. The rats were anesthetized with chloral hydrate after 10 h LPS administration, and then the blood samples were collected from the inferior vena cava and liver tissue was also obtained. Alanine aminotransferase (ALT) activity in serum and levels of 27 cytokines in liver tissue were tested. Correlation analysis of ALT and cytokines were performed using R 3.2.4 software. Results: Correlation analysis showed that there was a high positive correlation between ALT and macrophage inflammatory protein-1α (MIP-1α) or vascular endothelial growth factor (VEGF). There was a strong correlation among interleukin-18 (IL-18), interferon-inducible protein-10 (IP-10), interleukin-1α (IL-1α), and interleukin-1β (IL-1β). However, eotaxin regulated upon activation of normal T cell expression and secreted factor (RANTES), which has their own independence. Conclusion: Idiosyncratic liver injury induced by ZGW has a strong positive correlation with MIP-1α and VEGF, which provides a new experimental evidence for clinical medication safety and risk prevention of ZGW.
ABSTRACT
It is investigated that the hepatotoxicity of Polygonum multiflorum (PM)was attenuated by its processed products of nine times steaming and nine times sunning(RPM)based on immunological stress-mediated animal model by using metabolomics method. Sprague-Dawley(SD)rats were intragastrically administered with(5.4 g crude drug per kg body weight)of 50% alcohol extracts of PM and its processed products of nine times steaming and nine times sunning respectively or co-treated with non-toxic dose of lipopolysaccharide(LPS, 2.8 mg·kg-1)via tail vein injection. The plasma alanine aminotransferase(ALT)and aspartate aminotransferase(AST)activities were assayed and the isolated livers were evaluated for histopathological changes. Global metabolomics profiling, multivariate analysis and data base searching were performed to discover common differential metabolites for idiosyncratic liver injury. The results showed that co-treatment with non-toxic dose of LPS and PM could result in significant liver injury, indicated by the elevation of plasma ALT and AST activities, as well as obvious liver histologic damage; whereas RPM failed to induce detectable liver injury. Furthermore, 10 potential metabolomics biomarkers that differentially expressed in LPS/PM group compared with LPS/RPM without liver injury were identified by untargeted metabolomics, mainly involved ten pathways: sphingolipid metabolism, linoleic acid metabolism, taurine and hypotaurine metabolism, steroid hormone biosynthesis, galactose metabolism, steroid biosynthesis, metabolism of xenobiotics by cytochrome P450, pyrimidine metabolism, biosynthesis of unsaturated fatty acids, primary bile acid biosynthesis. This work illustrated the idiosyncratic hepatotoxicity of heshouwu and provided a metabolomic insight into diosyncratic liver injury of PM and RPM.
ABSTRACT
On basis of the idiosyncratic lipopolysaccharide(LPS)-mediated hepatotoxicity model, liver injury induced by Zhuangguguanjie wan(ZGW)was evaluated, and the mechanism was explored. Idiosyncratic hepatotoxicity model was established in rats by injecting LPS at a dosage of 2.8 mg·kg-1. Rats were randomly divided into the normal control group, LPS group, ZGW group and LPS+ZGW group. Alanine aminotransferase(ALT)and aspartate aminotransferase(AST)activities were analyzed in serum; pathological changes(HE staining)and the content of cytokines of liver were tested; and immune cell subpopulation ration were determined in blood and liver. Compared with the control group, the ZGW group and LPS group had no significant changes in ALT, AST and liver pathology(P> 0.05); while the ZGW+LPS group exhibited an elevation in ALT and AST(P< 0.05). Disorder of liver lobular arrangement and irregular island-like or massive necrosis of liver cells were observed in the group. Several cytokines in the liver were increased in LPS group and ZGW+LPS group(P< 0.05 or P< 0.01), and the level in ZGW+LPS group was higher than that of LPS group. Compared with the control group, the ratio of CD3+ T cell/lymphocyte of blood in LPS group was significantly decreased(P< 0.01); while the percentage of CD3+ T cells in the liver were significantly increased(P< 0.05). The contents of immune cells of blood had no significant changes between LPS group and ZGW+LPS group(P> 0.05). CD3+ T cell in the liver of ZGW+LPS group was significantly increased over the LPS group(P< 0.05). Aggregation or activity of CD3+ T cell was increased by ZGW combined with LPS. These results suggest that ZGW could promote T lymphocyte recruitment to liver under the immune activation state leading to inflammatory response, which may contribute to idiosyncratic liver injury.
ABSTRACT
To investigate the effects of peroxisome proliferator-activated receptor gamma(PPAR-γ)on the liver injury of Polygonum multiflorum, we established a model of immunological idiosyncrasy liver injury induced by lipopolysaccharide. The 70 Sprague-Dawley(SD)rats were randomly divided into control group, LPS group(2.8 mg·kg-1), PM group(crude drug, 2.16 g·kg-1), PPAR-γ agonist group(pioglitazone, 0.5 mg·kg-1), PM+LPS group(crude drug 2.16 g·kg-1, 2.8 mg·kg-1), PPAR-γ agonist+LPS group(0.5 mg·kg-1, 2.8 mg·kg-1)and PM+LPS+PPAR-γ agonist group(crude drug, 2.16 g·kg-1, 2.8 mg·kg-1, 0.5 mg·kg-1). The rats were orally given PM, once a day for consecutive 2 days. The control rats were given the same amount of distilled water. Liver injury was induced by intravenous injection of LPS. Sodium pentobarbital was injected intraperitoneally for anesthesia, and liver samples were collected together with blood. The plasma levels of alanine transaminase(ALT), aspartate aminotransferase(AST), tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), interleukin-6(IL-6)and interferon-γ(IFN-γ)were measured. Pathological changes and hepatocellular apoptosis were examined by liver biopsy, and immunohistochemical observation of liver tissue expression of PPAR-γ and NF-κB p65. A negative correlation was observed between the expression of PPAR-γ in hepatic tissue and liver injury of Polygonum multiflorum. PPAR-γ agonist significantly reduced the PM-induced idiosyncratic liver injury in rats according to serum ALT and AST(P < 0.05), reduced liver pathological injury and hepatocyte apoptosis, decreased serum TNF-α and other inflammatory cytokines(P < 0.05), liver tissue PPAR-γ expression, and inhibited expression of NF-κB p65(P < 0.05). The results suggest that the occurrence of immunological idiosyncrasy liver injury of PM is related to inhibition of the PPAR-γ pathway and elevation of inflammatory factors. PPAR-γ agonist can reverse the idiosyncratic liver injury induced by PM, and provide a reference for elucidating mechanism of idiosyncratic liver injury induced by Polygonum multiflorum.
ABSTRACT
Objective: To investigate the effects of crude and prepared Polygonum multiflorum (PM) to liver injury based on idiosyncratic hepatotoxicity model co-treated with lipopolysaccharide (LPS) at a non-hepatotoxic dose, aiming to compare the difference between them. Methods: The Sprague-Dawley (SD) rats were used by ig administration, respectively with 50% alcohol extracts from crude or prepared PM alone or co-treated noninjurious dose of bacterial lipopolysaccharide (LPS, 2.8 mg/kg) via tail vein injection. Blood was collected from the inferior vena cava into a sodium citrate-anticoagulant tube, and the liver was removed. The plasma samples separated from the collected blood were used for analysis on ALT and AST activities and the isolated livers were used for histopathological assessment. Results: Both crude and prepared PM were nontoxic on normal SD rats under the dose of 17.28 g/kg, when co-treated with LPS, the liver injury was found at the dose of 1.08 g/kg of crude PM, however, 8.64 g/kg of prepared PM. Conclusion: When processed, the maximum safe therapeutic dosage of PM would be enlarged from 1.08 g/kg of crude PM that is close to clinical dosage to 8.64 g/kg of prepared PM, which is four times larger than the crude one in LPS model. It shows that toxicity of PM could be reduced by processing.
ABSTRACT
Drug induced liver injury is an infrequent adverse effect of Verapamil. It has been reported in about ten cases since 1981. We report a case of a middle-aged woman who received Verapamil in a dose of 120mg/d for eight weeks, developing a mixed hepatocellular and cholestatic pattern of liver injury without any symptoms. There was no evidence of eosinophilia. Liver function tests returned quickly to normal levels after discontinuation. As in previously reported cases, we hypothesize that this is an idiosyncratic mechanism of liver injury.
La hepatotoxicidad es un efecto adverso infrecuente de verapamil, existiendo alrededor de 10 casos reportados desde 1981. Relatamos el caso de una mujer de edad media que presenta una alteración de las pruebas hepáticas con patrón mixto asociado al uso de verapamil 120 mg/d por 8 semanas sin síntomas asociados, la cual responde rápidamente a la suspensión de este fármaco. Al igual que en casos reportados previamente, planteamos como mecanismo de injuria una hepatotoxicidad idiosincrática por este medicamento.