Liver Inflammation In Rats Research Articles

Modulation of Xiongdanjiuxin pills on the gut-liver axis in high-fat diet rats

AimXiongdanjiuxin pill (XP) is a traditional Chinese medicine formula for the prevention and treatment of hyperlipidemia (HLP) and related complications. In this study, the gut-liver axis was used as the breakthrough point to analyze the therapeutic effect and potential mechanism of XP on HLP model rats and related complications. Main methodsWe used high-fat diet (HFD) to establish the HLP model of rats and treated them with XP. The 16S rRNA sequencing method was used to explore the effect of XP on the gut microbiota of HFD rats, and the effects of XP on ileum pathology, intestinal barrier and circulatory inflammation in HFD rats were also investigated. We further explored the molecular mechanism of XP treating liver inflammation in rats with HFD by regulating toll-like receptor 4 (TLR4) signaling. Key findingsWe found that XP could regulate the imbalance of gut microbiota in HFD rats, and up-regulate the expression of tight junction protein in intestinal epithelium of HFD rats, thereby improving the intestinal barrier damage and intestinal inflammatory response. In addition, XP could significantly reduce the levels of inflammatory cytokines in HFD rats, and inhibit TLR4 signaling pathway, thereby reducing liver inflammation in HFD rats. SignificanceXP can effectively improve the imbalance of gut-liver axis in hyperlipidemic rats and alleviate the inflammatory damage of liver. Its mechanism may be related to regulating the disorder of gut microbiota and inhibiting TLR4 signal pathway, so as to achieve the therapeutic effect on hyperlipidemic fatty liver in rats.

The toxic mechanism of 6:2 Cl-PFESA in adolescent male rats: Endocrine disorders and liver inflammation regulated by the gut microbiota-gut-testis/liver axis

In previous studies, 6:2 chlorinated polyfluorinated ether sulfonic acid (6:2 Cl-PFESA), a perfluorooctanesulfonate alternative, has been demonstrated to be toxic to mammals. However, the toxic mechanism of 6:2 Cl-PFESA in mammals is unknown. Herein, adolescent male rats were administered 50 μg/kg/Day 6:2 Cl-PFESA for 28 days (oral gavage) to estimate the toxicity of 6:2 Cl-PFESA and investigate its toxic mechanism. Significant changes in some hematological indicators (e.g., aspartate transaminase and neutrophils) and liver sections (inflammatory cell infiltration) indicated that 6:2 Cl-PFESA exposure caused rat hepatotoxicity. Six steroid hormones (e.g., testosterone, progesterone, and cortisol) in serum and thirteen genes in testicles (related to the pathway of steroid hormone biosynthesis) were significantly regulated in 6:2 Cl-PFESA-treated rats. This suggested that 6:2 Cl-PFESA induced rat endocrine disorders. Compared to the controls, the mean relative abundance of Ruminococcaceae, Pasteurellaceae, Micrococcaceae, and Desulfovibrionaceae was significantly regulated by 1.3-, 0.40-, 0.32-, and 3.2-fold in the 6:2 Cl-PFESA rats, respectively. The 6:2 Cl-PFESA treatment also significantly disturbed 47 gut metabolites (29 upregulated and 18 downregulated), mainly bile acids, short-chain fatty acids, and amino acids. In summary, 6:2 Cl-PFESA induced endocrine disorders and liver inflammation in rats by altering the gut microbiota-gut-testis/liver axis. This study first reveals the toxic mechanism of 6:2 Cl-PFESA in mammals through a multiomics approach and provides comprehensive insight into the toxic mechanism of 6:2 Cl-PFESA.

The ketogenic diet prevents steatosis and insulin resistance by reducing lipogenesis, diacylglycerol accumulation and protein kinase C activity in male rat liver

Obesity-associated insulin resistance plays a major role in the pathogenesis of non-alcoholic fatty liver disease (NAFLD). The accumulation of diacylglycerol (DAG), ceramides and inflammation are key factors that cause NAFLD. In recent years, the ketogenic diet (KD) has emerged as an effective non-pharmacological intervention for the treatment of NAFLD and other obesity-related metabolic disorders. What remains undetermined is how the KD affects DAG and ceramide content and insulin sensitivity in the liver. Thus, this research was designed to assess these variables, as well as glucose and fat metabolism and markers of inflammation in livers of rats exposed for 8weeks to one of the following diets: standard chow (SC), obesogenic high-fat, sucrose-enriched diet (HFS) or a KD. Despite having a higher fat content than the HFS diet, the KD did not cause steatosis and preserved hepatic insulin signalling. The KD reduced DAG content and protein kinase C-ε activity, but markedly increased liver ceramide content. However, whereas the KD increased ceramide synthase 2 (CerS2) expression, it suppressed CerS6 expression, an effect that promoted the production of beneficial very long-chain ceramides instead of harmful long-chain ceramides. The KD also enhanced the liver expression of key genes involved in mitochondrial biogenesis and fatty acid oxidation (Pgc-1α and Fgf21), suppressed inflammatory genes (Tnfα, Nf-kb, Tlr4 and Il6), and shifted substrate away from de novo lipogenesis. Thus, through multiple mechanisms the KD exerted anti-steatogenic and insulin-sensitizing effects in the liver, which supports the use of this dietary intervention to treat NAFLD. KEY POINTS: The accumulation of diacylglycerol (DAG), ceramides and inflammation are key factors that cause insulin resistance and non-alcoholic fatty liver disease (NAFLD). This study provides evidence that a ketogenic diet (KD) rich in fat and devoid of carbohydrate reduced DAG content and preserved insulin signalling in the liver. The KD shifted metabolism away from lipogenesis by enhancing genes involved in mitochondrial biogenesis and fatty acid oxidations in the liver. The KD also promoted the production of beneficial very long-chain ceramides instead of potentially harmful long-chain ceramides. Through multiple mechanisms, the KD exerted anti-steatogenic and insulin-sensitizing effects in the liver, which supports the use of this dietary intervention to treat NAFLD.

Protective Effect of Melatonin on Age-induced Changes in Circadian Clock and Inflammatory Profile in Rat Liver

Background: Aging is associated with a reduction in melatonin secretion and a deteriorating circadian system. In addition, aging is also defined by a chronic low-grade inflammation, also known as inflammaging. Sirtuin 1 (SIRT1) can affect clock function by binding with CLOCK/BMAL1 complexes and can be involved in the regulation of inflammation via deacetylation of high-mobility group box-1 (HMGB1) and inhibit the transcription of inflammation-related genes, NF-κB. Interestingly, melatonin, a hormone mainly secreted by the pineal gland, showed a variety of regulatory effects on circadian rhythm and inflammatory response in age-related liver disease. Objective: It is of interest to examine the effects of melatonin on age-induced changes in the levels of circadian clock protein and inflammation-related factors in rat liver. Materials and Methods: Rats were divided into 3 groups; young control group, aged control group, and aged rat treated with melatonin in drinking water (20 mg/L). After 2 months of melatonin administration, the liver tissues were collected. The levels of BMAL1, REV-ERBα, SIRT1, HMGB1, NF-κB, IL-6, and type I collagen, a marker of hepatic fibrosis, were analyzed by western blot analysis. Results: The present results showed that aged rats exhibited significantly downregulated BMAL1, REV-ERBα, and SIRT1 but upregulated HMGB1, NF-κB, IL-6, and type I collagen in rat liver compared with the young control group. Melatonin treated in aged rats significantly attenuated the age-induced downregulation of BMAL1, REV-ERBα, and SIRT1 and attenuated the age-induced increase in HMGB1, NF-κB, IL-6, and type I collagen protein expression. Conclusion: The present study suggested that the protective effect of melatonin on age-induced changes in clock-related inflammation in rat liver, including increasing the expression of clock and clock-controlled proteins, reducing the release of inflammatory profiles, and inhibiting fibrogenic response. These effects of melatonin may be involving the clock-related SIRT1/HMGB1/NF-κB axis. Keywords: Aging; Melatonin; Inflammaging; Clock gene; Liver; SIRT1; HMGB1; NF-κB

Protective Effect of Melatonin on Age-induced Changes in Circadian Clock and Inflammatory Profile in Rat Liver

Background: Aging is associated with a reduction in melatonin secretion and a deteriorating circadian system. In addition, aging is also defined by a chronic low-grade inflammation, also known as inflammaging. Sirtuin 1 (SIRT1) can affect clock function by binding with CLOCK/BMAL1 complexes and can be involved in the regulation of inflammation via deacetylation of high-mobility group box-1 (HMGB1) and inhibit the transcription of inflammation-related genes, NF-κB. Interestingly, melatonin, a hormone mainly secreted by the pineal gland, showed a variety of regulatory effects on circadian rhythm and inflammatory response in age-related liver disease. Objective: It is of interest to examine the effects of melatonin on age-induced changes in the levels of circadian clock protein and inflammation-related factors in rat liver. Materials and Methods: Rats were divided into 3 groups; young control group, aged control group, and aged rat treated with melatonin in drinking water (20 mg/L). After 2 months of melatonin administration, the liver tissues were collected. The levels of BMAL1, REV-ERBα, SIRT1, HMGB1, NF-κB, IL-6, and type I collagen, a marker of hepatic fibrosis, were analyzed by western blot analysis. Results: The present results showed that aged rats exhibited significantly downregulated BMAL1, REV-ERBα, and SIRT1 but upregulated HMGB1, NF-κB, IL-6, and type I collagen in rat liver compared with the young control group. Melatonin treated in aged rats significantly attenuated the age-induced downregulation of BMAL1, REV-ERBα, and SIRT1 and attenuated the age-induced increase in HMGB1, NF-κB, IL-6, and type I collagen protein expression. Conclusion: The present study suggested that the protective effect of melatonin on age-induced changes in clock-related inflammation in rat liver, including increasing the expression of clock and clock-controlled proteins, reducing the release of inflammatory profiles, and inhibiting fibrogenic response. These effects of melatonin may be involving the clock-related SIRT1/HMGB1/NF-κB axis. Keywords: Aging; Melatonin; Inflammaging; Clock gene; Liver; SIRT1; HMGB1; NF-κB

Lagerstroemia speciosa (L.) Pers., ethanolic leaves extract attenuates dapsone-induced liver inflammation in rats

Drug-induced liver injury is a common cause of acute liver failure. Dapsone is increasingly used in combination with rifampicin for the treatment of leprosy and also for several dermatological disorders. Clinically, abnormal liver function and focal bile duct destruction were reported after dapsone therapy. Lagerstroemia speciosa Pers., commonly known as Banaba has been traditionally used to treat various ailments including diabetes and obesity due to its antioxidant and anti-inflammatory efficacies. This study investigated the hepatoprotective effect of ethanolic banaba leaves extract (EBLE) against dapsone-induced hepatotoxicity in rats. Dapsone (30 mg/kg, i.p.) was administered twice daily for 30 days. In separate groups, rats were post-treated orally with EBLE (250 and 500 mg/kg) and silymarin (100 mg/kg) once daily for 30 days after dapsone administration. The marker enzymes of hepatotoxicity, oxidative stress markers, inflammatory markers and histopathology of liver were done. HPTLC analysis confirmed the presence of 12.87 µg of corosolic acid per mg of EBLE. Dapsone administration-induced significant (p < 0.001) elevation of marker enzymes of hepatotoxicity in serum. This treatment also increased lipid peroxidation (p < 0.001) and pro-inflammatory markers (tumor necrosis factor-alpha, transforming growth factor-beta, and nuclear factor kappa-B) expressions (p < 0.001) and decreased antioxidants (p < 0.001) such superoxide dismutase, catalase and glutathione in the liver tissue. All these abnormalities were significantly (p < 0.001) mitigated after EBLE (500 mg/kg) and silymarin post-treatments. The results of this study suggest that silymarin and EBLE can be used for dapsone-induced hepatotoxicity.

Kinsenoside Alleviates 17α-Ethinylestradiol-Induced Cholestatic Liver Injury in Rats by Inhibiting Inflammatory Responses and Regulating FXR-Mediated Bile Acid Homeostasis.

Cholestasis is an important predisposing factor of liver diseases, such as hepatocyte necrosis, liver fibrosis and primary biliary cirrhosis. In this study, we aimed to investigate the effects of Kinsenoside (KD), a natural active ingredient of Anoectochilus roxburghii, on estrogen-induced cholestatic liver injury in Sprague-Dawley rats and the underlying mechanism. The rats were randomly divided into six groups: control group, model group, low-dose KD group (50 mg/kg body weight, KD-L), medium-dose KD group (100 mg/kg body weight, KD-M), high-dose KD group (200 mg/kg body weight, KD-H) and ursodeoxycholic acid group (40 mg/kg body weight, UDCA). 17α-Ethinylestradiol (EE) was used to establish an experimental animal model of estrogen-induced cholestasis (EIC). The results demonstrated that KD alleviated liver pathologic damage, serum biochemical status and inhibited hepatocellular microstructure disorder and bile duct hyperplasia in EE-induced cholestatic rats. Mechanically, KD alleviated EE-induced cholestatic liver injury by inhibiting inflammatory responses and regulating bile acid homeostasis. Concretely, KD reduced the expression of IL-1β and IL-6 by inhibiting NF-κB p65 to suppress EE-mediated inflammation in rat liver. KD enhanced the expression of FXR and inhibited EE-mediated reduction of FXR in vitro and in vivo. It was the potential mechanism that KD mitigates cholestasis by increasing efflux and inhibiting uptake of bile acids via FXR-mediated induction of bile salt export pump (BSEP) and reduction of Na+-dependent taurocholate cotransport peptide (NTCP) to maintain bile acid homeostasis. Moreover, KD repressed the bile acid synthesis through reducing the expression of synthetic enzyme (CYP7A1), thereby normalizing the expression of metabolic enzyme (SULT2A1) of bile acid. In conclusion, our results revealed that KD may be an effective drug candidate for the treatment of cholestasis.

Lauric acid ameliorates lipopolysaccharide (LPS)-induced liver inflammation by mediating TLR4/MyD88 pathway in Sprague Dawley (SD) rats

BackgroundLipopolysaccharide (LPS) is an endotoxin that leads to inflammation in many organs, including liver. It binds to pattern recognition receptors, that generally recognise pathogen expressed molecules to transduce signals that result in a multifaceted network of intracellular responses ending up in inflammation.AimIn this study, we used lauric acid (LA), a constituent abundantly found in coconut oil to determine its anti-inflammatory role in LPS-induced liver inflammation in Sprague Dawley (SD) rats. MethodMale SD rats were divided into five groups (n = 8), injected with LPS and thereafter treated with LA (50 and 100 mg/kg) or vehicle orally for 14 days. After fourteen days of LA treatment, all the groups were humanely killed to investigate biochemical parameters followed by pro-inflammatory cytokine markers; tumour necrosis factor-α (TNF-α), interleukin-6 (IL-6), and IL-1β. Moreover, liver tissues were harvested for histopathological studies and evaluation of targeted protein expression with western blot and localisation through immunohistochemistry (IHC). ResultsThe study results showed that treatment of LA 50 and 100 mg/kg for 14 days were able to reduce the elevated level of pro-inflammatory cytokines, liver inflammation, and downregulated the expression of TLR4/NF-κB mediating proteins in liver tissues. ConclusionThese findings suggest that treatment of LA has a protective role against LPS-induced liver inflammation in rats, thus, warrants further in-depth investigation through mechanistic approaches in different study models.

The ameliorating effect of silymarin against vancomycin-induced apoptosis and inflammation in rat liver

The ameliorating effect of silymarin against vancomycin-induced apoptosis and inflammation in rat liver

Curcumin-loaded PLA-PEG copolymer nanoparticles for treatment of liver inflammation in streptozotocin-induced diabetic rats.

This report focused on loading curcumin (CUR) drug into biodegradable Polylactide-poly(ethylene glycol) (PLA-PEG) copolymer nanoparticles as an effective anti-inflammatory agent in vivo to overcome the limitations resulted from the free CUR. By a simple nano-emulsification technique, hydrophobic CUR was loaded into hydrophobic polymer's segments and stabilized by cationic surfactant. They were then characterized by DLS, TEM, and SEM techniques providing monodispersed and spherical nanoparticles with an average diameter of 117 nm and high surface charge of +35 mV. Thereafter, they were orally administrated into five groups of rats, typically, control (healthy rats), streptozotocin (STZ)-induced diabetic rats, diabetics treated with free CUR, diabetics treated with PLA-PEG NPs, and diabetics treated with CUR-encapsulated PLA-PEG NPs. Next, complete blood analyses were assessed including alanine aminotransferase (ALT), aspartate aminotransferase (AST), and nuclear factor kappa B (NF-ҡB), reduced glutathione (GSH), malondialdehyde (MDA), nitric oxide (NO), cyclooxygenase (COX-2), Peroxisome proliferator-activated receptors (PPAR-γ) and transforming growth factor-β1 (TGF-β1). The obtained results demonstrated that diabetes initially produced liver inflammation in rats manifested by leveraging the mean levels of serum AST, ALT inducing oxidative stress resulting in a clear increase in the levels of hepatic MDA and NO concomitant with a remarkable decrease in GSH. Moreover, diabetes significantly increased serum NF-ҡB, hepatic COX-2 and TGF-β1, while highly reduced hepatic PPAR-γ. In contrast, both CUR free and CUR-encapsulated NPs ameliorated the negative changes in diabetes but CUR-encapsulated NPs showed more pronounced treated effect than free CUR. In addition, histopathological investigations were performed on the liver tissues of all groups, showing a mitigation in inflammation while treating with CUR-NPs.

Genistein can ameliorate hepatic inflammatory reaction in nonalcoholic steatohepatitis rats

Genistein plays an active role in improving nonalcoholic fatty liver disease (NAFLD). This study is designed to investigate the effect of genistein on liver inflammation in rats with nonalcoholic steatohepatitis (NASH). Forty SPF male SD rats were randomly divided into normal group, model group, genistein low-dose group (0.1% wt/wt) and high-dose group (0.2% wt/wt) with 10 rats in each group. After 12 weeks’ feeding, liver tissues and serum samples of rats were taken, and HE staining was used to perform pathological examination of liver tissues, then the degree of inflammatory infiltration was observed and NAFLD activity score(NAS) was calculated. With corresponding kits, several indicators were detected, namely, serum triglyceride (TG), total cholesterol (TC), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), liver TC and TG, and serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood glucose and serum endotoxin. The levels of tumor necrosis factor (TNFα) in liver and insulin in blood of rats were detected by enzyme linked immunosorbent assay (ELISA), then the HOMA-IR index was calculated. Immunohistochemistry staining was used to observe the expression level of TLR4 protein and the RT-PCR was used to detect Tlr4 mRNA expression in liver tissue. The results showed that genistein could reduce TLR4 protein and gene expression, decrease the endotoxin and TNFα, alleviate the inflammatory reaction and make the indicators detected in blood and liver stay near normal in NASH rats. In conclusion, genistein can ameliorate hepatic inflammatory reaction in nonalcoholic steatohepatitis rats.

Acute treatment with Mangifera indica L. leaf extract attenuates liver inflammation in rats fed a cafeteria diet.

This study investigates the acute anti-inflammatory activity of Mangifera indica L. leaf extract and mangiferin in the liver of rats fed a cafeteria diet. This study was a randomized longitudinal experimental study. The animals were divided into three groups - Control: cafeteria diet (CD); Extract: CD + leaf extract (250 mg kg-1); and Mangiferin: CD + mangiferin (40 mg kg-1). Body weight and food intake were measured every week. On day eight, mRNA and protein expression of inflammatory markers were evaluated in the liver. Also, liver weight, SOD activity and malondialdehyde concentration were measured. Treatment for only eight days with mango leaf extract and mangiferin increased SOD activity. Mangiferin intake increased the mRNA expression of PPAR-α and HSP72. The leaf extract treatment enhanced PPAR-α mRNA expression. Mangiferin and leaf extract consumption caused a lower concentration of NFκB (p65) in nuclear extracts, and greater IL-10 mRNA and protein levels. This study highlights the potential of acute treatment with mango leaf extract and mangiferin to prevent liver inflammation caused by fat-rich diets. These results indicate a new use for a product that has low cost, is found in great amounts, and is not routinely used.

Hepatoprotective effect of ultrasonicated ginseng berry extract on a rat mild bile duct ligation model

BackgroundThe Panax ginseng berry extract (GBE) is well known to have an antidiabetic effect. The aim of this study is to evaluate and investigate the protective effect of ultrasonication-processed P. ginseng berry extract (UGBE) compared with GBE on liver fibrosis induced by mild bile duct ligation (MBDL) model in rats. After ultrasonication process, the composition ratio of ginsenoside in GBE was changed. The component ratio of ginsenosides Rh1, Rh4, Rg2, Rg3, Rk1, Rk3, and F4 in the extract was elevated. MethodsIn this study, the protective effect of the newly developed UGBE was evaluated on hepatotoxicity and neuronal damage in MBDL model. Silymarin (150 mg/kg) was used for positive control. UGBE (100 mg/kg, 250 mg/kg, 500 mg/kg), GBE (250 mg/kg), and silymarin (150 mg/kg) were orally administered for 6 weeks after MBDL surgery. ResultsThe MBDL surgery induced severe hepatotoxicity that leads to liver inflammation in rats. Also, the serum ammonia level was increased by MBDL surgery. However, the liver dysfunction of MBDL surgery–operated rats was attenuated by UGBE treatment via myeloid differentiation factor 88-dependent Toll-like receptor 4 signaling pathways. ConclusionUGBE has a protective effect on liver fibrosis induced by MBDL in rats through inhibition of the TLR4 signaling pathway in liver.

Pre-and postnatal exposition to fluorides induce changes in rats liver morphology by impairment of antioxidant defense mechanisms and COX induction

IntroductionFluorides are common in the environment and are absorbed mostly in the stomach and gut, it can easily move through cell membranes and its accumulation can cause harmful effects in skeletal and soft tissues. One of the most important F- accumulation sites is the liver.The aim of this study was to determine whether F- can cause inflammation in rat liver by affecting the activity of antioxidant enzymes and changes in the synthesis of prostaglandin E2 (PGE2) and thromboxane B2 (TXB2). Materials and methodsAn in vivo model of prenatal and postnatal exposure to sodium fluoride (NaF) was used to carry out the experiment. Animals from control group received tap water to drink, while animals exposed to F- received drinking water containing NaF, 50 mg/L. In serum and liver we analyzed F- concentration, in liver - antioxidant enzymes activity, PGE2 and TXB2 concentration and immunolocalization of COX1 and COX2 proteins were measured. ResultsWe observed significant changes in F- concentration only in liver. The results of this study showed that F- affects antioxidant enzymes activity, COX2 protein expression and PGE2 synthesis in liver. Also, in some regions of the liver of rats exposed to F-, the hepatocytes were diffusely altered, with changes resembling microvesicular steatosis. ConclusionChronic exposure to F− during development causes an accumulation of this element in the liver and changes in antioxidant enzymes activity and cyclooxygenase expression. Long term exposure to this element is toxic to the liver and can cause disturbances in its homeostasis.

Hepatoprotective role of Solenostemma argel growing in Egypt on ethanol induced oxidative damage in rats

Abstract Objective The objective of the current study is to investigate the protective effect of Solenostemma argel (S. argel) treatment on ethanol (EtOH)-induced hepatotoxicity in rat liver. Methods Forty adult male albino rats were divided into four groups as control, S. argel (100 mg/kg), EtOH (3 g/kg) and S. argel + EtOH groups. S. argel was given 1 h prior to EtOH by oral gavage for 28 days. Results The results showed that, administration of EtOH caused a significant decrease (p&lt;0.05) in serum total protein and albumin, whereas ALT and AST and lipid peroxidation (LPO) were increased following EtOH treatment. S. argel treatment significantly (p&lt;0.05) ameliorated the previous parameters. Protein carbonyl, reduced glutathione and LPO levels were significantly (p&lt;0.05) increased along with glutathione-S-transferase activity was decreased (as markers of oxidative stress) in EtOH treated rat liver. Previous oxidative stress was attenuated significantly by S. argel treatment. Moreover, in EtOH group, tumor necrosis factor-α (TNF-α) and nitric oxide (NO) contents and cytochrome P450 2E1 activity were significantly increased in liver tissues showing oxidative organ damage. Co-administration of S. argel with EtOH significantly reversed the inflammation in rat livers. Conclusion S. argel had a hepatoprotective role against EtOH-induce oxidative stress and inflammation in rat liver.

Mechanism of Rhei Radix et Rhizoma combined with Scutellariae Radix on liver inflammation in rats with endotoxin disease based on p38MAPK pathway

To explore the mechanism of Rhei Radix et Rhizoma combined with Scutellariae Radix in regulatory lipopolysaccharide (LPS)-induced liver inflammation in rats with endotoxin blood, 50 male SD rats were selected and randomly divided into blank group, model group, dexamethasone group, herbal pair high dose group, herbal pair low dose group, with 10 in each group. Rats in each were given preventive drugs for 7 consecutive days. At 0.5 h after the final administration, the model was built through the tail vein injection with LPS (5 mg•kg⁻¹). Then, animal anal temperatures were determined and recorded once every 0.5 h. The rats were killed at 4 h after the modeling to determine spleen thymus coefficient. ELISA method was used to detect cytokines interleukin-1β (IL-1β) and tumor necrosis factor-alpha (TNF-α) in liver tissues. The colorimetric method was applied in determination of oxidation nitrogen (NO) content in liver tissues. Western Blot method was adopted to detect Toll-like receptor protein 4, p38MAPK p38MAPK, phosphorylated p38MAPK (p-p38MAPK) and nitric oxide synthase (iNOS) protein expressions. The results showed that compared with the blank group, in the model group, TLR4 protein expression, iNOS protein expression and p38 phosphorylation expression, IL-1β, NO and TNF-α content increased significantly in liver tissues (P<0.05 or P<0.01). And compared with the model group, the herbal pair high dose group showed significantly reduction in IL-1β, NO and TNF-α expressions in rat liver tissues (P<0.05 or P<0.01), down-regulation in iNOS protein expression and p38 phosphorylation expression in rat liver tissues (P<0.05), but without significant up-regulation in TLR4 protein. Low-dose herbal pair can significantly reduce IL-1β and NO expression in model rat liver tissues (P<0.01), significantly down-regulate iNOS protein expression (P<0.01), with a slight down-regulation in phosphorylation of p38 but no statistical significance, and no reduction in TLR4 expression. In conclusion, the compatibility of Rhei Radix et Rhizoma combined with Scutellariae Radix may reduce the expression of iNOS protein and the release of inflammatory cytokines IL-1β, NO and TNF-α by decreasing p38 protein phosphorylation expression and blocking p38MAPK signaling pathways, so as to alleviate the inflammation reaction and protect the liver.

Caloric Restriction Prevents Lead-Induced Oxidative Stress and Inflammation in Rat Liver

The aim of the present study is to investigate the effects of caloric restriction on liver of lead-administered rat. Male Sprague-Dawley rats were randomly divided into two groups: Ad libitum fed group (AL, free access to normal rat chow) and caloric restriction group (CR, fed 65% of AL animals' food intake). After 6 weeks, half of the animals of each group were injected lead acetate and the other half were injected saline. Liver tissue samples were collected at the end of the experiments. Glutathione peroxidase (GPx), superoxide dismutase (SOD), malondialdehyde (MDA), and tumor necrosis factor (TNF-α) were measured in the tissue extracts. Histological studies were also performed. Our results showed that lead administrations (not saline injections) reduced liver SOD and GPx and increased MDA and TNF-α in AL animals, but in the CR animals lead injections did not significantly change the measured parameters. The histological studies supported the biochemical findings. We concluded that 65% CR may prevent lead-induced oxidative stress and inflammation in rat liver.

Cationic peptide mR18L with lipid lowering properties inhibits LPS-induced systemic and liver inflammation in rats

The cationic single domain peptide mR18L has demonstrated lipid-lowering and anti-atherogenic properties in different dyslipidemic mouse models. Lipopolysaccharide (LPS)-mediated inflammation is considered as one of the potential triggers for atherosclerosis. Here, we evaluated anti-inflammatory effects of mR18L peptide against LPS-mediated inflammation. First, we tested the efficacy and tolerance of 1, 2.5 and 5mg/kg mR18L in normolipidemic rats stimulated with 5mg/kg LPS. LPS and then mR18L were injected in different intraperitoneal regions. By 2h post LPS, mR18L inhibited LPS-mediated plasma TNF-α elevation at all doses, with the effect being stronger for 2.5mg/kg (P<0.05 vs. 1mg/kg, non-significant vs. 5mg/kg). In a similar model, 2.5mg/kg mR18L reduced LPS-mediated inflammation in the liver, as assessed by microscopic examination of liver sections and measurements of iNOS expression in the liver tissue. In plasma, 2.5mg/kg mR18L decreased levels of TNF-α and IL-6, decreased endotoxin activity and enhanced HDL binding to LPS. In another similar experiment, mR18L administered 1h post LPS, prevented elevation of plasma triglycerides by 6h post LPS and increased plasma activity of anti-oxidant enzyme paraoxonase 1, along with noted trends in reducing plasma levels of endotoxin and IL-6. Surface plasmon resonance study revealed that mR18L readily binds LPS. We conclude that mR18L exerts anti-endotoxin activity at least in part due to direct LPS-binding and LPS-neutralizing effects. We suggest that anti-endotoxin activity of mR18L is an important anti-inflammatory property, which may increase anti-atherogenic potential of this promising orally active lipid-lowering peptide.

Hot Water ExtractedLycium BarbarumandRehmannia GlutinosaInhibit 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 (CCl(4))-induced liver injury in rats. Male Sprague-Dawley rats were randomly divided into: normal diet + peritoneal injection of olive oil (control), normal diet + CCl(4) injection (CCl(4)), 1 × HE (0.05% HE for each) + CCl(4) (1 × HE), and 3 × HE (0.15% HE for each) + CCl(4) (3 × HE) groups. Rats were injected with 40% CCl(4) 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 CCl(4) 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 CCl(4)-induced liver injury.

Dioscorea villosa (wild yam) induces chronic kidney injury via pro-fibrotic pathways

Dioscorea villosa (wild yam) rhizome extract is a medicinal herb that is commonly used to treat symptoms of menopause and rheumatoid arthritis. We had evidence from previous in vitro experiments that this extract is toxic and pro-fibrotic in renal cells and aimed to test whether this occurs in vivo. Sprague-Dawley rats received 0.79 g/kg/d D. villosa extract in their food or no treatment over 7, 14 and 28d ( n = 4 per group). Kidney and liver tissues were collected for protein extraction and Western immunoblots or fixed for special histologic stains, immunohistochemistry (IHC) and microscopy. Collagen deposition was assessed using Masson’s trichrome staining and morphometry. Macrophage infiltration (ED-1), epithelial-to-mesenchymal transdifferentiation or activation of fibroblasts (vimentin, α-SMA), and pro-fibrotic growth factors (TGFß1, CTGF) were assessed using IHC. Protein expression levels of the pro-inflammatory cytokine, TNF-α, the pro-fibrotic transcription factor, NFκB, a measure of oxidative stress (heme oxygenase-1), α-SMA, vimentin and TGFß1 were determined. Results showed that kidneys of the treated animals had significantly increased collagen, vimentin, TGFβ1, NFκB, EDI, CTGF and α-SMA by 28d. In the liver, there was increased ED-1 and TGFβ1 in the centrilobular zone at 28d in treated animals. In conclusion, there was no acute reno- or hepato-toxicity associated with administration of D. villosa. However, there was an increase in fibrosis in the kidneys and in inflammation in livers of rats consuming D. villosa for 28 days. Long term supplementation with D. villosa may be best avoided, especially in people with compromised renal function and in those who need to take other drugs which may alter kidney function.