Increased Serum Alanine Aminotransferase Research Articles

The mechanism of Psoralen and Isopsoralen hepatotoxicity as revealed by hepatic gene expression profiling in SD rats.

The main bioactive components of Fructus psoraleae, such as psoralen and isopsoralen, are known to be hepatotoxic. However, its underlying mechanism is to be elucidated. To address this, SD rats were randomly divided into control group, 60mg/kg psoralen group and 60mg/kg isopsoralen group. Blood was collected to detect serum biochemical indices. RNA was extracted from liver samples, and then, cDNA gene expression profiles were analysed. Psoralen administration significantly up-regulated serum AST (aspartate aminotransferase) while addition of isopsoralen increased serum ALT (alanine aminotransferase), AST, TBA (total bile acid) and TG (total triglyceride) levels. A total of 172 differentially expressed genes (DEGs) were acquired between psoralen group and control group while 884 DEGs were screened between isopsoralen group and control group. Chemical Carcinogenesis and Metabolism of Xenobiotics by Cytochrome P450 were the two most significantly enriched pathways as revealed by DEGs. Liver was the most impacted organ, and endoplasmic reticulum was the most impacted organelle in subcellular level. Finally, some kinds of cancers and cytochrome p450 oxidoreductase deficiency were predicted. Taken together, psoralen and isopsoralen might cause hepatotoxicity mainly through cytochrome P450 metabolism of xenobiotics. Furthermore, Cyp1a1, Cyp1a2, Gstm1 and Akr7a3 worked as key genes in hepatotoxicity. Moreover, endoplasmic reticulum was the main target subcellular structure in hepatotoxicity induced by psoralen and isopsoralen.

Comparative study on dose-toxicity-effect of Tripterygium Glycosides Tablets and Tripterygium wilfordii Tablets on CIA model rats

The aim of this paper was to compare the properties of Tripterygium Glycosides Tablets and Tripterygium wilfordii Tablets from dose-effect-toxicity on type Ⅱ collagen-induced arthritis( CIA) in rats. SD rats were randomly divided into eight groups,including normal group,model group,Tripterygium Glycosides Tablets groups( 1 times equivalent dose 0.009 g·kg-1,4 times equivalent dose 0.036 g·kg-1,16 times equivalent dose 0.144 g·kg-1),Tripterygium wilfordii Tablets groups( 1 times equivalent dose 0.007 5 mg·kg-1,4 times equivalent dose 0.030 mg·kg-1,16 times equivalent dose 0.120 mg·kg-1). Beginning on the first immunization,Tripterygium Glycosides Tablets and Tripterygium wilfordii Tablets administered intraperitoneally once a day. After the second immunization,the symptoms such as redness and swelling of joints were observed,and the clinical score and incidence of arthritis were evaluated. HE and Masson staining were used to examine the histopathological changes of joints. The expression level of anti-type Ⅱ collagen antibody Ig G in serum was detected by ELISA,routine testing of blood components,the concentration of ALP( alkaline phosphatase),ALT( alanine aminotransferase),AST( aspartate aminotransferase),GGT( gamma-glutamyltransferase),TBi L( total bilirubin),CRE( creatinine) and UREA( urea) in serum were detected by enzymatic assay. The rate of sperm deformity in the epididymis was evaluated under light microscope. The extent of damage to the testis and ovarian tissue was assessed by HE staining. Tripterygium Glycosides Tablets and Tripterygium wilfordii Tablets attenuated the inflammation,redness,swelling and deformity of joints and reduced the clinical score and incidence of arthritis in CIA rats. Meanwhile,it also exhibited obvious reduction in all pathological features such as joint synovitis,pannus,cartilage erosion and bone destruction. Tripterygium Glycosides Tablets and Tripterygium wilfordii Tablets reduced Ig G in a dose-dependent manner,and Tripterygium Glycosides Tablets is better than Tripterygium wilfordii Tablets( P<0.05 or P<0.01). The high doses of Tripterygium Glycosides Tablets and Tripterygium wilfordii Tablets could significantly increase the organ coefficient of liver and spleen and reduced RBC and HGB in CIA rats( P<0.01),and severity leading to death. Gastric mucosal injury and morphological changes of liver and kidney were not observed in CIA rats of Tripterygium Glycosides Tablets and Tripterygium wilfordii Tablets treatment group. The 4 and 16 times doses of Tripterygium Glycosides Tablets and Tripterygium wilfordii Tablets could significantly increase serum ALT,GGT and decrease CRE( P<0.05 or P<0.01). Tripterygium Glycosides Tablets and Tripterygium wilfordii Tablets could increase the sperm deformity rate and damage the testicular seminiferous tubules of CIA male rats. Severity increased with dose and time increasing. The effect of Tripterygium Glycosides Tablets( 16 times) is more significant than Tripterygium wilfordii Tablets( 16 times). Tripterygium Glycosides Tablets and Tripterygium wilfordii Tablets significantly delayed onset of arthritis and inhibited the paw edema and arthritic score. Tripterygium Glycosides Tablets and Tripterygium wilfordii Tablets also caused male reproductive damage,high dose affected hematopoiesis,and maximum dose leading to death. Tripterygium Glycosides Tablets and Tripterygium wilfordii Tablets all depended on dose-effect-toxicity manner. Anti-arthritis effect of Tripterygium Glycosides Tablets is better than Tripterygium wilfordii Tablets,but the toxicity of Tripterygium Glycosides Tablets maximum dose is more obvious. The relevant conclusions of our study will provide experimental references for clinical rational use of drugs,and further clinical studies are needed to confirm our conclusions.

Adenovirus‑mediated overexpression of bone morphogenetic protein‑9 promotes methionine choline deficiency‑induced non‑alcoholic steatohepatitis in non‑obese mice

Liver inflammation and macrophage infiltration are critical steps in the progression of non-alcoholic fatty liver to the development of non-alcoholic steatohepatitis. Bone morphogenetic protein-9 is a cytokine involved in the regulation of chemokines and lipogenesis. However, the function of bone morphogenetic protein-9 in non-alcoholic steatohepatitis is still unknown. The present study hypothesized that bone morphogenetic protein-9 may contribute to steatohepatitis in mice fed a methionine choline deficiency diet (MCD). C57BL/6 mice overexpressing bone morphogenetic protein-9 and control mice were fed the MCD diet for 4 weeks. Liver tissue and serum samples were obtained for subsequent measurements. Bone morphogenetic protein-9 overexpression exacerbated steatohepatitis in mice on the MCD diet, as indicated by liver histopathology, increased serum alanine aminotransferase activity, aspartate transaminase activity, hepatic inflammatory gene expression and M1 macrophage recruitment. Although bone morphogenetic protein-9 overexpression did not affect the expression of pro-fibrogenic genes, including Collagen I (α)1 or matrix metalloproteinase (MMP) 9, it did upregulate the expression of transforming growth factor-β and plasminogen activator inhibitor 1, and downregulated the expression of MMP2. The above results indicate that bone morphogenetic protein-9 exerts a pro-inflammatory role in MCD diet-induced non-alcoholic steatohepatitis.

Synergistic curative effect of Boswellic acid and Cisplatin against Diethyl nitrosamine -induced hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is a primary malignancy of hepatocyte. Boswellic acids are the effective components of gum resin of Boswellia serrata, which have anti-inflammatory properties. Cisplatin is used as a chemotherapeutic agent for treatment of HCC. The present study aims to evaluate the role of Boswellic acid in HCC treatment and as a chemo-sensitizer for Cisplatin treatment of HCC in rats that is induced by orally administration of Diethyl nitrosamine (DEN). Sixty rats were divided in to sex groups, 10 rats each, group 1: normal control group, group 2: cisplatin (1.5 mg/Kg b.wt), group 3: received the Diethyl nitrosamine (DEN) (20 mg/kg b.wt), group 4: received DEN as in group 3 and then treated with cisplatin, group 5: received DEN as in group 3 and then treated with Boswellic acid (500 mg/kg b.wt) and group 6: received DEN as in group 3 and then treated with cisplatin and Boswellic acid. After 10 weeks animals were sacrificed and liver tissue were removed for Histopathological and tissue parameters examination. A significant increase in serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) activity, a significant decrease in serum albumin concentration. Also a significant increases in liver transforming growth factor-beta 1 (TGF-b1), B-cell lymphoma 2 (Bcl-2) and nuclear factor kappa β (NF-kβ) concentrations and a significant decrease in Caspase.3 activity were recorded in DEN-treated rats while treatment with Boswellic acid and Cisplatin reduced the severity of HCC and enhanced histopathological findings, these result suggesting the efficacy of Boswellic acid supplementation as an anti-HCC.

Liver toxicity assessments in rats following sub-chronic oral exposure to copper nanoparticles

BackgroundThe widespread use of nano-copper as a feed additive in the absence of toxicological studies has potential risks to humans and animals. Toxicity studies on nano-copper in animals usually exposure from the respiratory tract, however, it is necessary to study the oral exposure toxicity of nano-copper to understand its risks as a feed additive.ResultsCurrently the hepatotoxicity and mechanism of nano-copper after sub-choronic oral exposure at equivalent doses of 50, 100, and 200 mg/kg/day were evaluated in rats; micro-copper and Cu ions were used as controls. Nano-copper (200 mg/kg) increased serum alanine aminotransferase and aspartate aminotransferase significantly, further promoting hepatic oxidative stress, inflammation, and corresponding histopathological changes, and exhibited significant dose-dependent changes. Nano-copper also decreased the level of nuclear receptors, resulting in significant reductions in mRNA, protein, and activity of hepatic CYP450 enzymes. The molecular mechanisms responsible for these toxic effects involved the signaling pathway of NF-κB, MAPK, and STAT5.ConclusionsNano-copper caused strong hepatic toxicity by inducing oxidative stress and inflammation. The decreased drug metabolism enzymes lead to nano-copper–drug interaction that provoked the concerns on animal-derived food safety.

Receptor-Interacting Serine/Threonine-Protein Kinase 3 (RIPK3)–Mixed Lineage Kinase Domain-Like Protein (MLKL)–Mediated Necroptosis Contributes to Ischemia-Reperfusion Injury of Steatotic Livers

Increased hepatic ischemia-reperfusion (IR) injury in steatotic livers is a major reason for rejecting the use of fatty livers for liver transplantation. Necroptosis is implicated in the pathogenesis of fatty liver diseases. Necroptosis is regulated by three key proteins: receptor-interacting serine/threonine-protein kinase (RIPK)-1, RIPK3, and mixed-lineage kinase domain-like protein (MLKL). Here, we found that marked steatosis of the liver was induced when a Western diet was given in mice; steatosis was associated with the inhibition of hepatic proteasome activities and with increased levels of key necroptosis-related proteins. Mice fed a Western diet had more severe liver injury, as demonstrated by increases in serum alanine aminotransferase and necrotic areas of liver, after IR than did mice fed a control diet. Although hepatic steatosis was not different between Mlkl knockout mice and wild-type mice, Mlkl knockout mice had decreased hepatic neutrophil infiltration and inflammation and were protected from hepatic IR injury, irrespective of diet. Intriguingly, Ripk3 knockout or Ripk3 kinase-dead knock-in mice were protected against IR injury at the late phase but not the early phase, irrespective of diet. Overall, our findings indicate that liver steatosis exacerbates hepatic IR injury via increased MLKL-mediated necroptosis. Targeting MLKL-mediated necroptosis may help to improve outcomes in steatotic liver transplantation.

Cerium Oxide Nanoparticles Attenuate Oxidative Stress and Inflammation in the Liver of Diethylnitrosamine-Treated Mice.

The catalytic activity of cerium oxide nanoparticles (CeO2NPs) is responsible for its application as an antitumor agent. This activity may be due to its ability to switch between III and IV oxidation states thereby conferring pro- and antioxidant properties. This study was designed to assess the hepatoprotective potential of CeO2NPs in male BALB/c mice administered diethylnitrosamine (DEN). Thirty-six mice were divided equally into six groups and treated intraperitoneally with normal saline (control), DEN (200mg/kg) alone, CeO2NPs 1 (100μg/kg) + DEN (200mg/kg), CeO2NPs 2 (200μg/kg) + DEN (200mg/kg), CeO2NPs 1 alone, and CeO2NPs 2 alone. Animals were pretreated with CeO2NPs daily for eight consecutive days, while DEN was administered 48h before the animals were sacrificed. Administration of DEN caused a significant increase in serum alanine aminotransferase (ALT) and urea by 51% and 96%, respectively. Markers of oxidative stress (malondialdehyde) and inflammation (nitric oxide and myeloperoxidase) in hepatic tissues of DEN-treated mice were increased by 60%, 16%, and 38%, respectively. The activities of hepatic superoxide dismutase, catalase, glutathione peroxidase, glutathione-S-transferase, and level of reduced glutathione were significantly decreased in DEN-treated mice by 50%, 123%, 23%, 419%, and 78%, respectively. In addition, DEN increased the expression of hepatic Bcl2 and COX-2, while p53, Bax, and iNOS were mildly expressed. Pretreatment with CeO2NPs attenuated the activities of antioxidant enzymes and expression of Bcl2 and COX-2. Overall, CeO2NPs confers protection from DEN-induced liver damage via antioxidative activity.

Treatment of a Mouse Model of Cholestasis with a Thyromimetic Improves Biliary Injury But Exacerbates Hepatocyte Injury

Chronic cholestasis results from bile secretory defects or impairment of bile flow, and there are few effective medical therapies available. Thyroid hormone T3 and synthetic thyroid hormone receptor agonists are known to cause induction of hepatocyte proliferation during liver regeneration. However, whether these drugs have therapeutic benefits in cholestatic liver disease is unknown. In this study, we administered GC‐1, a thyromimetic that preferentially acts through the TRb receptor found predominantly in liver, to Mdr2 knockout (KO) mice, which is a commonly used model of sclerosing cholangitis characterized by bile acid (BA) regurgitation, periductular inflammation, and fibrosis. We determined Mdr2 KO mice fed 5mg/kg GC‐1 diet had decreased bilirubin, liver to body weight ratios, serum alkaline phosphatase, but increased serum alanine aminotransferase and aspartate aminotransferase compared to KO mice fed normal diet as early as 1 week on diet. Histologically, KO mice on GC‐1 diet had decreased ductular response, less bridging fibrosis, and fewer SOX9 positive hepatocytes compared to KO on normal diet. Although total liver BA were higher in KO mice fed GC‐1 diet for 2 weeks compared to normal diet, they normalized to KO levels at 4 weeks of diet. To elucidate the mechanism of increased BA accumulation and liver injury, we examined expression of BA transporters and detoxification enzymes. KO mice on GC‐1 diet had decreased bilirubin transport and detoxification genes, such as Mrp3, Cyp2b10, and Oatp4, compared to KO mice on normal diet, with the net result being retention of BA in the hepatocyte. Affymetrix gene array data also indicates that KO mice on GC‐1 have normalized bile acid synthesis compared to WT expression levels. Thus, GC‐1 reduces cholangiocyte injury during cholestasis by inducing retention of BA in hepatocytes, causing injury to the hepatocytes; this occurs through as‐yet unknown mechanisms that are upstream of BA transporters and biosynthesis enzymes and appears to be b‐catenin independent.Support or Funding InformationNational Institute of Biomedical Imaging and Bioengineering of the National Institute of Health under Award Number T32EB0010216UPP Academic Foundation Award 1R01DK103775This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Characterization and biological activities of polysaccharides from artificially cultivated Phellinus baumii.

The polysaccharide (PPB-2) was extracted and purified from the fruiting body of artificially cultivated Phellinus baumii by an acid-chlorite delignification method. The in vitro hypoglycemic and in vivo hepatoprotective activities of PPB-2 were investigated. FT-IR spectra, NMR spectral analysis, and scanning electron microscopy were used to investigate the structural characterization of PPB-2. PPB-2 exhibited higher α-glucosidase and α-amylase inhibitory activities and showed significant inhibitory effects against the diffusion of glucose. Furthermore, the administration of PPB-2 to mice significantly prevented the increase of serum alanine aminotransferase and aspartate aminotransferase activities, as well as the increase in total cholesterol, triglycerides, and total bilirubin, and hepatic malondialdehyde levels, and PPB-2 enhanced the activities of superoxide dismutase and catalase, as well as the glutathione levels. The results suggest that PPB-2 may be suitable as a functional food with hypoglycemic and hepatoprotective activities.

Association between PNPLA3 rs738409 polymorphism and nonalcoholic fatty liver disease (NAFLD) susceptibility and severity: A meta-analysis.

Objective:This meta-analysis is to investigate the relationship between the patatin-like phospholipase domain containing 3 (PNPLA3) rs738409 polymorphism and the susceptibility and severity of nonalcoholic fatty liver disease (NAFLD).Methods:Chinese Journal Full-text Database, Wanfang Database, VIP Database, and PubMed Database were subjected to case-control study retrieving, from January 2008 to December 2014. Following key words were used: fatty liver, PNPLA3, and rs738409 gene or variants or polymorphism or alleles. Meta-analysis was performed based on the retrieved articles.Results:In total 65 studies were first retrieved according to the key words, and finally 21 studies with 14,266 subjects were included. Meta-analysis showed that PNPLA3 rs738409 polymorphism exerted strong influence not only on fatty liver but also on the histological injury. PNPLA3 rs738409 [G] allele was a risk factor for NAFLD (GG vs CC, OR = 4.01, 95% CI 2.93–5.49; GC vs CC, OR = 1.88, 95% CI 1.58–2.24). PNPLA3 gene variant was significantly associated with the increased serum alanine aminotransferase (ALT) levels (GG vs CC, standardized mean difference = 0.47, 95% CI 0.14–0.81). In addition, nonalcoholic steatohepatitis (NASH) was more frequently observed in G allele carriers (GG vs CC, OR = 3.24, 95% CI 2.79–3.76; GC vs CC, OR = 2.14, 95% CI 1.43–3.19).Conclusion:PNPLA3 rs738409 polymorphism is not only a factor significantly associated with the susceptibility of NAFLD, but also related to the susceptibility of aggressive diseases.

Possible Protective Role of Panax Ginseng on Cisplatin-Induced Hepatotoxicity in Adult Male Albino Rats (Biochemical and Histological Study).

Background:Cisplatin is one of the most effective chemotherapy antineoplastic drugs. Panax ginseng is a well-known medicinal herb and has a long history of medicinal use as a tonic to promote health.Aim:This work aimed to study the effect of ginseng on the liver damage induced by cisplatin in rats. It included biochemical and histological investigations.Materials and Methods:Twenty adult rats were divided into four equal groups. Group I served as control. Group II received ginseng orally (100 mg/kg/day) for 4 weeks. Group III animals were injected intraperitoneally with cisplatin in three equal doses (each 3.3 mg/kg) daily for 3 consecutive days. Group IV animals received ginseng together with cisplatin by the same previously mentioned methods and doses. Rats were sacrificed after 4 weeks, and blood samples and liver tissues were collected for biochemical and histological examinations.Results:Cisplatin-induced liver damage manifested biochemically by an increase in serum alanine aminotransferase and aspartate aminotransferase. Histologically, hepatocytes appeared with vacuolated cytoplasm and small dark-stained nuclei with dilatation of blood sinusoids as well as marked accumulation of collagen fibers around enlarged portal tracts. Administration of ginseng together with cisplatin improved the hepatic dysfunctions and damage caused by cisplatin.Conclusion:Ginseng has a protective role in the amelioration of cisplatin-induced hepatotoxicity.

THE POSSIBLE AMELIORATIVE EFFECT OF VITAMIN C AND ARTEMISIA JUDAICA EXTRACT AGAINST TOXICITY INDUCED BY ZINC OXIDE NANOPARTICLES IN MALE ALBINO RATS

Recently, many biological fields including medicine, personal care products, sunscreens and food additives, also many industrial domains such as ceramics, rubber, paints and alloys, have been extensively applied by Zinc oxide nanoparticles (ZnO NPs), as it has the ability to pass through the cell membrane easily and even pass through blood-brain barrier. They are highly reactive and may cause oxidative stress, which induces serious damages in DNA and protein structures forming mutation. So, the current study was designed to assess the possible mitigating effect of Artemisia judaica (Art. j.), vitamin C (vit. C) or their co-administration against ZnO NPs–induced hepatorenal toxicity in male rats. Eighty adult male rats were divided into 8 groups: the 1st (control group) with distilled water for 45 days; the 2nd (twin 80 group) with twin 80 for 45 days; the 3rd (ZnO NPs group), rats were gavaged daily with 10 mg/kg body weight (b.wt) of ZnO NPs for 15 days; the 4th (vit. C group), rats were gavaged daily with 100 mg/kg b.wt of vit. C for 30 days; the 5th (Art. j. group), rats were gavaged daily with 150 mg/kg b.wt of Art. j. extract for 30 days; the 6th (ZnO NPs + vit. C group), rats were gavaged daily with 10 mg/kg b.wt of ZnO NPs for 15 days then gavaged daily with 100 mg/kg b.wt of vit. C for 30 days; the 7th (ZnO NPs + Art. j. group), rats were gavaged daily with 10 mg/kg b.wt of ZnO NPs for 15 days then gavaged daily with 150 mg/kg b.wt of Art. j. for 30 days; the 8th (ZnO NPs + vit. C and Art. j. group), rats were gavaged daily with 10 mg/kg b.wt of ZnO NPs for 15 days then gavaged daily with Art. j. and vit. C co-administration for 30 days. ZnO NPs group showed a high significant increase in serum alanine aminotransferase (ALT), aspartate amino transferase (AST), alkaline phosphatase enzyme (ALP) and gamma glutamyltransferase(GGT) activities, serum total and direct bilirubin, creatinine, urea and uric acid levels, and blood urea nitrogen (BUN) and malondialdhyde (MDA) levels. While, it showed a high significant decrease in serum albumin, protein and reduced glutathione (GSH) levels, serum superoxide dismutase (SOD), glutathione peroxidase (GPX) and catalase (CAT) activities, as well as blood hemoglobin (Hb), red blood cells (RBCs), white blood cells (WBCs) and packed cell volume (PCV) as compared with control group. ZnO NPs groups that were treated by vit. C or Art. j. showed partial ameliorative effect against ZnO NPs-induced hepatorenal toxicity with convergent degree, but the efficacy of Art.j. was better than vit. C. ZnO NPs group that was treated by vit. C and Art. j. co-administration showed marked significant curative effects against ZnO NPs- induced hepatorenal toxicity.

Akt2 Mediates Glucocorticoid Resistance in Acute Lymphoblastic Leukemia through FoxO3a/Bim Axis and Serves As a Direct Target for Resistance Reversal

Background: Glucocorticoids (GCs) are common components in chemotherapeutic protocols for acute lymphoblastic leukemia (ALL). A major obstacle in GC therapy, however, is the gradual acquisition of apoptotic resistance in ALL cells repeatedly treated with these hormones. Previous reports indicate that 15-30% of pediatric ALL samples are resistant to GCs, while in refractory childhood ALL, the prevalence of GC resistance is as high as 70%. Identification of specific molecular mechanisms driving resistance to GC and targeting downstream molecules may lead to the development of new therapeutic strategies.Results: FoxO transcription factor FoxO3a has been shown to regulate apoptosis in lymphocytes. Unphosphorylated FoxO3a can be upregulated by dexamethasone (DEX), which subsequently translocates into the nucleus, upregulates Bim expression and induces apoptosis. In our current study, we cultured a GC-sensitive T-ALL cell line CCRF-CEM with a specific concentration of DEX over multiple passages and obtained a highly resistant cell line, designated CEM-DR. We observed this T-ALL cell line acquires resistance to DEX-mediated killing through abnormal activation of Akt, resulting in inhibition of the FoxO3a/Bim pathway (A). Pharmacologic inhibition of Akt with Akt inhibitor IV effectively restores sensitivity to DEX of CEM-DR by enhancing the FoxO3a/Bim pathway, but shows significant hepatotoxicity with increased serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in vivo (B). Common hematologic toxicities and hepatic toxicities with increased AST and ALT of Akt inhibitor have also been reported in the treatment of solid tumors in humans, partially limiting their clinical applicability (Becher OJ et al, Pediatr Blood Cancer 2017). There are two closely related, highly conserved homologues of Akt: Akt-1 and -2, which differ in enzyme function. Our study aimed to investigate the potential role of Akt isoforms Akt1 and Akt2 in the mechanism of GC resistance and explore a more direct and specific target for resistance reversal.By western blot analysis we observed the expression of Akt2 in intrinsically resistant T-ALL cells Jurkat was significantly higher than that in sensitive CCRF-CEM cells, while Akt1 expression in aforementioned cell lines was similar. The expression of Akt2 also increased synchronously with the increase of half maximal inhibitory concentration (IC50) of DEX in CEM-DR cells, further indicating that Akt2 expression increases in secondarily resistant lymphocytes. A significantly elevated expression of Akt2 not Akt1 were shown in relapsed/refractory ALL patients when compared with the newly diagnosed patients. To detect if Akt2 was able to directly interact with FoxO3a, co-immunoprecipitation assay was employed. MYC-FoxO3a was co-transfected with Flag-Akt1 or Flag-Akt2 into HEK293T cells. The result demonstrated the more presence of Flag-AKT2 than Flag-Akt1 in MYC-FoxO3a immunoprecipitates, suggesting that AKT2 as the major regulator directly interacting with FoxO3a. Reciprocal immunoprecipitation experiments confirmed the closer association between Flag-Akt2 and MYC-FoxO3a (C). Then we used siRNA to down-regulate Akt1 or Akt2 expression in resistant T-ALL cells; we observed GC-induced apoptosis increased significantly (D) after down-regulation of Akt2 expression, along with the expression of p-FoxO3a decreased (E) .To examine the therapeutic role of Akt isoform specific inhibitors, we treated resistant T-ALL cell with A-674563 (Akt1 inhibitor), CCT128930 (Akt2 inhibitor) or Akti1/2 (Akt1/2 inhibitor). Selective inhibition of Akt2 with CCT128930 more significantly enhances the FoxO3a/Bim signaling pathway, increases DEX-mediated killing of resistant T-ALL cells (F) and effectively reverses GC resistance than Akt1 inhibitor in vitro and in vivo. When exploring the potential influence on viscera by Akt isoform inhibitors in vivo, we found Akt2 inhibitor did not significantly influence hematopoiesis, cardiac and renal functions. Notably, Akt2 inhibitor did not show hepatic toxicities of increased serum ALT, AST and total bilirubin which appear in Akt1 or Akt1/2 inhibition.Conclusions: Akt2 might serve as a more direct and specific kinase mediating GC resistance through FoxO3a/Bim-signaling pathway in ALL, and targeting Akt2 with CCT128930 may be explored as a promising therapeutic strategy for resistance reversal. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Inhibition of Triggering Receptor Expressed on Myeloid Cells 1 Ameliorates Inflammation and Macrophage and Neutrophil Activation in Alcoholic Liver Disease in Mice.

Alcoholic liver disease (ALD) is characterized by macrophage and neutrophil leukocyte recruitment and activation in the liver. Damage‐ and pathogen‐associated molecular patterns contribute to a self‐perpetuating proinflammatory state in ALD. Triggering receptor expressed on myeloid cells 1 (TREM‐1) is a surface receptor that amplifies inflammation induced by toll‐like receptors (TLRs) and is expressed on neutrophils and monocytes/macrophages. We hypothesized that TREM‐1 signaling contributes to proinflammatory pathway activation in ALD. Using an in vivo ALD model in mice, we tested the effects of ligand‐independent TREM‐1 inhibitory peptides that were formulated into human high‐density lipoprotein (HDL)‐mimicking complexes GF9‐HDL and GA/E31‐HDL. As revealed in vitro, macrophages endocytosed these rationally designed complexes through scavenger receptors. A 5‐week alcohol feeding with the Lieber‐DeCarli diet in mice resulted in increased serum alanine aminotransferase (ALT), liver steatosis, and increased proinflammatory cytokines in the liver. TREM‐1 messenger RNA (mRNA) expression was significantly increased in alcohol‐fed mice, and TREM‐1 inhibitors significantly reduced this increase. TREM‐1 inhibition significantly attenuated alcohol‐induced spleen tyrosine kinase (SYK) activation, an early event in both TLR4 and TREM‐1 signaling. The TREM‐1 inhibitors significantly inhibited macrophage (epidermal growth factor‐like module‐containing mucin‐like hormone receptor‐like 1 [F4/80], clusters of differentiation [CD]68) and neutrophil (lymphocyte antigen 6 complex, locus G [Ly6G] and myeloperoxidase [MPO]) markers and proinflammatory cytokines (monocyte chemoattractant protein 1 [MCP‐1], tumor necrosis factor α [TNF‐α], interleukin‐1β [IL‐1β], macrophage inflammatory protein 1α [MIP‐1α]) at the mRNA level compared to the HDL vehicle. Administration of TREM‐1 inhibitors ameliorated liver steatosis and early fibrosis markers (α‐smooth muscle actin [αSMA] and procollagen1α [Pro‐Col1α]) at the mRNA level in alcohol‐fed mice. However, the HDL vehicle also reduced serum ALT and some cytokine protein levels in alcohol‐fed mice, indicating HDL‐related effects. Conclusion: HDL‐delivered novel TREM‐1 peptide inhibitors ameliorate early phases of inflammation and neutrophil and macrophage recruitment and activation in the liver and attenuate hepatocyte damage and liver steatosis. TREM‐1 inhibition represents a promising therapeutic approach for further investigations in ALD.

Observations on the Activities of Certain Serum Enzymes and Levels of Some Biochemical Parameters in Spent Horses with Liver Damage

Clinical biochemistry tests are one of the tests designed to assess the functional status of the liver in domestic animals and man. This study evaluated the activities of some serum enzymes and the level of some biochemical parameters in horses with acute and chronic liver damage. This study was carried out in Nigerian horses presented for sale at the Obollo Afor horse depot, Udenu Local Government Area, Enugu State Nigeria. A total of 100 adult Nigerian horses of either sex were studied once a week for a period of 24 weeks. Serum biochemistry parameters indicated for the assessment of liver function were evaluated following standard procedures. These parameters include: Alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), total proteins, albumin, globulin, total cholesterol, total bilirubin and urea. Based on the outcome of the serum biochemistry evaluation, the horses were assigned into 3 groups namely: apparently healthy horses (Group 1), horses with biochemical markers of acute liver damage (Group 2) and horses with biochemical markers of chronic liver damage (Group 3). The mean serum ALT, AST, ALP and total bilirubin values of group 2 horses were significantly (p &lt; 0.05) higher than those of groups 1 and 3 horses. The mean serum albumin value of group 2 horses was significantly (p &lt; 0.05) lower than that of groups 1 and 3 horses. There was no significant (p &gt; 0.05) difference in the mean serum urea value of group 2 and 3. A significantly (p &lt; 0.05) lower serum creatinine value was observed in group 2 horses. There were no significant (p &gt; 0.05) differences in the mean serum total protein, globulin and cholesterol values across the groups. In the spent horses, acute liver damage was characterized by increased serum ALT, AST and ALP activities, and decrease in creatinine level, hyperbilirubinemia, slight hypoalbuminemia, normal total protein, globulin and cholesterol levels while chronic liver damage was characterized by decreased serum ALT activity, normal serum AST and ALP activities, decreased serum urea level, normal total protein, albumin, globulin, total bilirubin and cholesterol levels.

Dietary Green Tea Extract Prior to Spinal Cord Injury Prevents Hepatic Iron Overload but Does Not Improve Chronic Hepatic and Spinal Cord Pathology in Rats.

Spinal cord injury (SCI) disrupts autonomic regulation of visceral organs. As a result, a leading cause of mortality in the SCI population is metabolic dysfunction, and an organ central to metabolic control is the liver. Our recent work showed that rodent SCI promotes Kupffer cell (hepatic macrophage) activation, pro-inflammatory cytokine expression, and liver steatosis. These are symptoms of nonalcoholic steatohepatitis (NASH), the hepatic manifestation of metabolic syndrome, and these pre-clinical data replicate aspects of post-SCI human metabolic dysfunction. Because metabolic profile is highly dependent on lifestyle, including diet, it is likely that lifestyle choices prior to injury influence metabolic and hepatic outcomes after SCI. Therefore, in this study we tested if a diet rich in green tea extract (GTE), a known hepatoprotective agent, that began 3 weeks before SCI and was maintained after injury, reduced indices of liver pathology or metabolic dysfunction. GTE treatment significantly reduced post-SCI hepatic iron accumulation and blunted circulating glucose elevation compared with control-diet rats. However, GTE pre-treatment did not prevent Kupffer cell activation, hepatic lipid accumulation, increased serum alanine transaminase, or circulating non-esterified fatty acids, which were all significantly increased 6 weeks post-injury. Spinal cord pathology also was unchanged by GTE. Thus, dietary GTE prior to and after SCI had only a minor hepatoprotective effect. In general, for optimal health of SCI individuals, it will be important for future studies to evaluate how other lifestyle choices made before or after SCI positively or negatively impact systemic and intraspinal outcomes and the overall metabolic health of SCI individuals.

Chemical characteristics, antioxidant capacities and hepatoprotection of polysaccharides from pomegranate peel

This proposed work aimed to investigate the chemical characteristic, antioxidant capacities and hepatoprotection effect of pomegranate peel polysaccharides (PPP) on CCl4-induced oxidative damage in mice. PPP was identified as the acidic heteropolysaccharides by HPLC methods. In vitro test showed that PPP had excellent reducing power and scavenging effects against free radicals. Administration of PPP (50, 100 and 200 mg/kg·bw) in mice before the injection of CCl4 could observably antagonize the increased serum alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase and hepatic malonaldehyde level in CCl4-induced mice, especially administrated with 200 mg/kg·bw of PPP. Hepatic enzymatic activities of total superoxide dismutase, glutathione peroxidase, catalase and non-enzymatic activity of glutathione were markedly increased at high dosage of PPP, respectively. In addition, histopathological observation of liver further proved these biochemical characteristics. Therefore, it can be concluded that PPP exhibits strong protective effects against CCl4-induced liver injury in mice.

INFLUENCE OF SELENIUM ON MONO SODIUM GLUTAMATE AS FOOD ADDITIVE INDUCED HEPATOTOXICITY AND TESTICULAR TOXICITY IN ADULT ALBINO RATS

Background: Mono sodium glutamate (MSG) is commonly used as a flavor enhancer. Many studies showed toxic effects of MSG on different organs. Selenium (Se) is reported to possess a strong antioxidant property. The aim of this work was to evaluate the role of selenium on hepatotoxicity, testicular toxicity induced by MSG in adult albino rats. Material and Methods: study included 55 male albino rats for 8 weeks. Rats were divided into four groups, group I(control group) consisted of 22 rats equally and randomly subdivided into 2 subgroups: Ia (-ve control group), Ib (+ve control group) received distilled water. Group II (Se group) consisted of 11 rats received sodium biselenite at a dose of (0.5ml/kg/day/orally) dissolved in distilled water. Group III (MSG treated group) consisted of 11rats received MSG at a dose of (830mg/kg/day/ orally) dissolved in distilled water. Group IV (MSG and Se treated group) consisted of 11 rats treated in the previous doses. After 8 weeks, the rats were submitted to estimate the serum levels of alanine transaminase (ALT) & gamma glutamyle transferase (GGT) levels and testosterone hormone level. Then the anesthetized rats were sacrificed & specimens from the liver and testes were taken for determination of histopathological study, immunohistochemical staining for caspase 3, oxidative stress markers [malondialdehyde (MDA) & glutathione peroxidase (GPx)] and sperm count. Results showed in MSG group significantly increased serum ALT, GGT & significantly decreased in serum testosterone, sperm count with marked histopathological changes in the liver and testis and caspase 3 activities was significantly increased in testicular and liver tissue. Also, it significantly increased MDA level and decreased GPx activity in (liver and testicular tissues). Administration of Se along with MSG produced partial improvement of hepatic and testicular morphological changes, reduction in caspase 3 expression with beneficial effect on liver and testicular parameters, In addition, it decreased MDA level and increased GPx activity in (liver and testicular tissues). In conclusion, the results confirmed the hepatic and testicular toxicity of MSG through oxidative stress. Selenium has partial protective effects against MSG induced hepatic, testicular toxicity through its anti oxidant and its anti apoptotic effects.

The hydroethanolic Litchi chinensis leaf extract alleviate hepatic injury induced by carbon tetrachloride (CCl4) through inhibition of hepatic inflammation

Various medicinal plants are traditionally used in a hepatoprotective manner, like, for example, the Litchi chinensis leaf infusion that is employed in Chinese medicine as liver tonics to strengthen hepatic functioning. In this context, the present study was designed to evaluate the hepatoprotective and acute toxicological effects of hydroethanolic L. chinensis leaf extract in HepG2 cells and mice. Specifically, the cytotoxicity and hepatoprotective activities of L. chinensis leaf extract were evaluated in HepG2 cells and in vivo against carbon tetrachloride (CCl4)-induced acute liver injury. The administration of CCl4 in mice provokes cell swelling, loss of sinusoid capillary spaces and structural disarrangement of the hepatic lobe, apoptosis and leukocyte infiltration. Further, CCl4 evokes an increase in serum alanine aminotransferase (ALT), tumor necrosis factor (TNF) and interleukin-6 (IL-6) levels in hepatic tissue. However, Silymarin, the positive control, and the L. chinensis extract were able to restore the viability of cells treated with CCl4 at all concentrations evaluated, reduced the inflammatory parameters, TNF and IL-6, reestablished hepatic tissue morphology and did not induce acute toxicological alterations. The data obtained underscore that the extract from L. chinensis leaves features hepatoprotective activity, corroborating with ethnopharmacological use, and does not lead to acute toxicological effects.

Palmitate-induced lipotoxicity is crucial for the pathogenesis of nonalcoholic fatty liver disease in cooperation with gut-derived endotoxin

Although previous studies have indicated important roles of palmitate, a saturated fatty acid, in the pathogenesis of nonalcoholic fatty liver disease (NAFLD), it remains unclear how palmitate contributes to inflammation and fibrosis in the liver. Administration of palmitate in high fat diet (HFD)-fed but not basal diet (BD)-fed mice resulted in an increase in serum alanine aminotransferase (ALT) levels. Surprisingly, combined administration of very low dose lipopolysaccharide in palmitate-treated mice led to a marked increase in serum ALT levels despite BD-fed conditions. Administration of palmitate alone in BD-fed mice caused inflammatory cell infiltration and liver fibrosis mediated by the toll-like receptor 4 pathway without ALT elevation. In addition, a significant correlation between serum free fatty acid levels and liver fibrosis stage was observed in patients with NAFLD. These results indicate that palmitate may play crucial roles in the pathogenesis of NAFLD in the presence of gut-derived endotoxin.