Histological Changes In Liver Research Articles

HISTOPATHOLOGICAL EFFECT OF CHLORPYRIFOS PESTICIDE ON LIVER AND KIDNEY OF FRESH WATER FISH CATLA CATLA

All the natural water bodies are continuously being contaminated with toxic chemicals from industrial, agricultural and domestic activities. Chlorpyrifos is one of the most widely used organophosphate pesticides. Excessive application of pesticides from agricultural fields contaminate aquatic medium, resulting in serious damage to non-target species, including fish. The fish as a bioindicator of aquatic medium plays an important role in the monitoring of water pollution. Histopathology is an important tool for evaluating the action of any toxicant at tissue level. The focus of the present study is to measure the histological changes in liver and kidney of the fresh water fish Catla catla exposed to sub lethal concentrations of chlorpyrifos 1/15th of the 96 hour LC50 values for the period of 10 and 20 days. The fish exposed to chlorpyrifos showed mild vacuolar hepatocytes, dilation of central vein, vaculor degenerative changes, mild hyperaemia and severe hyperaemia of central vein in the liver tissue slide. The changes observed in kidney slide due to the impact of chlorpyrifos toxicity are hemorrhage, degenerative changes in between tubules, constriction of primary tubule, vacuolardegenerative changes, necrosis in epithelial lining of proximal tubules, chlorpyrifos compound induced.

Tianhuang formula ameliorates non-alcoholic fatty liver diseases in type 2 diabetic mice through CRLS1-ATF3/ChREBP pathway

ObjectiveTianhuang Formula (THF) is a hospital formula summarized by Professor Jiao Guo's 30 years of clinical experience. Some studies have shown that it can alleviate dyslipidemia in the body. The purpose of this study is to confirm whether THF can improve non-alcoholic fatty liver diseases (NAFLD) in type 2 diabetic mice induced by high-fat diet (HFD)/streptomycin (STZ) and to clarify its potential mechanism. MethodsAfter induction of diabetes, mice were administrated with THF (60 ​mg/kg or 120 ​mg/kg) once daily for 10 weeks. Blood glucose (FBG), glucose tolerance, and insulin resistance (IR) were assayed by oral glucose tolerance test (OGTT) and insulin tolerance test (ITT). Blood lipids, alanine transaminase (ALT), and aspartate transaminases (AST) were detected. Serum fasting insulin (INS) and adiponectin (APN) levels were measured using ELISA. Histological changes in liver and pancreatic islets were observed by H&E staining, followed by Oil Red O staining for liver lipid quantification and periodic acid-Schiff (PAS) staining to detect glycogen accumulation. Western blotting detected the levels of fatty cardiolipin synthase 1 (CRLS1), transcription factor activator 3 (ATF3), and carbohydrate-responsive element binding protein (ChREBP) in the liver. The mRNA transcripts of hepatic inflammatory factors, lipogenesis and lipolysis-related genes, and gluconeogenic enzyme-phosphoenolpyruvate carboxykinase (PEPCK), CRLS1, ATF3, and ChREBP mRNA levels were evaluated by RT-qPCR. ResultsTHF restored impaired glucose tolerance and insulin resistance, respectively. There was an improvement in HFD/STZ-induced liver and islet damage, high serum HDL-C and ANP levels, and a significant decrease in FBG, total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), FFA, INS, ALT, and AST, and lipid droplet counts in the T2DM mice treated with THF. CREBBP binding protein ATF3 mediated the insulin resistance signaling pathway, which regulated glucose and lipid metabolism in the liver. THF upregulated CRLS1, and ChREBP downregulated the expression of downstream ATF3 in the liver. RT-qPCR analysis also systemically indicated that THF suppressed the pathway and key regulators related to inflammation, lipid accumulation, and gluconeogenesis. ConclusionOur findings demonstrated that THF ameliorated lipid profile and attenuated liver steatosis in T2DM mice through CRLS1-ATF3/ChREBP pathway activation.

Effect of Annona muricata Plant Extract on the Multiple-organ Histology of Chloramphenicol-induced Leukemia in Wistar Albino Rats

This study was designed to show the histological changes of heart, kidney and liver of Wistar rats treated with Annona muricata plant extract upon oral administration of chloramphenicol. Thirty-five Wistar rats, divided into five groups of seven rats each was used for this study. The groups were normal control, untreated control and treated control administered 500mg/kg, 750mg/kg and 2250mg/kg body weight of Annona muricata plant extract respectively. This study lasted for four weeks. Organ specimens were obtained from each group to evaluate histological changes. This study revealed no histological changes in the heart muscle of Wistar rats given chloramphenicol. In the kidney, oral administration of chloramphenicol caused chronic pyelonephritis while the administration of this drug caused congestion in the liver. The administration of Annona muricata plant extract as treatment in 500mg/kg, 750mg/kg and 2250mg/kg doses helped reverse these damages in the kidney and liver tissues. Although rats treated with 750mg/kg body weight of Annona muricata exhibited inflammation in the heart muscle with an anaemia.

Limited role for hyperammonemia in the progression of diet-induced nonalcoholic steatohepatitis.

To determine whether hyperammonemia has a direct impact on steatohepatitis in mice fed with a high-fat diet (HFD). Male C57BL/6 mice were divided into two groups receiving either chow diet or HFD. After 12-week NASH modeling, hyperammonemia was induced by intragastric administration of ammonium chloride solution (NH4 Cl) or liver-specific carbamoyl phosphate synthetase 1 (Cps1) knockdown. In vitro experiments were performed in HepG2 cells induced by free fatty acid (FFA) and NH4 Cl. NH4 Cl administration led to increased levels of plasma and hepatic ammonia in NASH mice. NH4 Cl-induced hyperammonemia did not influence liver histological changes in mice fed with HFD; however, elevated plasma cholesterol level, and an increasing trend of liver lipid content were observed. No significant effect of hyperammonemia on hepatic inflammation and fibrosis in NASH mice was found. In vitro cell experiments showed that NH4 Cl treatment failed to increase the lipid droplet content and the expressions of de novo lipogenesis genes in HepG2 cells induced by FFA. The knockdown of Cps1 in HFD-fed mice resulted in elevated plasma ammonia levels but did not cause histological change in the liver. Our study revealed a limited role of ammonia in aggravating the progression of NASH. Further studies are needed to clarify the role and mechanism of ammonia in NASH development.

Hepato-protective potential of Choline and DHA supplements in rats exposed to tobacco particulate matter-A histological study

Introduction: Smokeless tobacco is known to induce liver damage by decreasing its detoxifying capability. Chronic exposure to tobacco particulate matter in various forms jeopardizes the normal function of vital organs including the liver. The tobacco rolled in tendu leaf; known as ‘bidi’ is an unfiltered cigarette having tobacco content that is different from those used in cigarettes and is referred to as 'bidi tobacco'. Bidi smoking or chronic exposure to bidi tobacco causes multi-organ diseases. Choline and docosahexaenoic acid (C & DHA) are dietary components known to have hepato-protective action. But the combined action of choline and DHA on tobacco particulate-induced liver damage is largely unknown. The present study was designed to assess the hepato-protective potential of choline and DHA supplements to rat dams and pups exposed to tobacco particulate matter. Liver histological changes were analyzed from groups of Wistar rat dams and their pups [Unexposed Normal controls (NC) and those exposed to different tobacco particulate matters namely, tendu leaf smoke, tobacco dust, and bidi smoke] with or without supplementation of both choline and DHA. Results: Hepatocytic morphological architecture showed non-alcoholic fatty changes in all rats exposed to tobacco particulate matter and more so visibly higher in tobacco dust exposed groups compared to the same in age-matched NC group. These changes were ameliorated in rats supplemented with choline & DHA and exposed to tobacco particulate matter. Conclusion: Chronic exposure of mothers and their offspring to tobacco particulates causes non-alcoholic fatty liver disease with microstructural changes. Dietary supplementation of choline and DHA to rats exposed to tobacco particulate matter provides hepato-protection and ameliorates the hepatocytic morphological changes.

Protective Effect of Andropraphis Paniculata Aqueous Extract (EAAp) Against Isoniazid and Rifampicin-Induced Rat Liver Damage

Isoniazid (INH) and rifampicin (RIF) are first-line antituberculosis drugs (OAT) in tuberculosis treatment that are used for at least 6 months. The use of OAT has been associated with toxic reactions in the liver and causes hepatitis. This study aimed to determine the effect of an aqueous extract of Andrographis paniculata (EAAp) on liver damage induced by INH and RIF. Method: Male Sprague-Dawley rats weighing 250–300g were divided into 5 groups, each consisting of 6 mice. Animals were given isoniazid and rifampicin at 100 mg/kg, respectively, to induce liver damage, silymarin (25 mg/kg) for the positive control group, and Ap extract at doses of 200mg and 300 mg/kg for the test group. All treatments were given orally once daily for 28 days. Measurement of serum aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), bilirubin, and liver histopathology levels was carried out to determine the effect of EAAp on liver damage by INH and RIF. Results: Rats treated with INH+RIF were hepatotoxic, as evidenced by increased serum ALT, AST, and ALP activity, total bilirubin levels, and histopathological changes in the liver. Administration of Ap extract doses of 200 mg/kg and 300 mg/kg significantly decreased liver biochemical and histological changes caused by OAT. Conclusions: EAAp has a protective effect against hepatotoxic-induced INH and RIF in animal models.

Composition and hepatoprotective effect of geopropolis of Melipona subnitida

Background: Geopropolis produced by bees of the subfamily Meliponinae (Hymenoptera, Apidae) is used in Brazilian popular medicine for the treatment of different diseases. The aim of the present work was to evaluate the hepatoprotective potential of the hydroethanolic extract from geopropolis (HPE) produced by Melipona subnitida Ducke. Additionally, the chemical composition of HPE and antioxidant activity were also evaluated. Methods: The hepatoprotective potential of HPE was evaluated by serum biochemical profile and macroscopic and histological examination of the liver of 24 Wistar rats (Rattus norvegicus), distributed in four experimental groups: G1 (negative control), G2 (HPE), G3 (HPE plus acetaminophen), G4 (acetaminophen - positive control). The HPLC-DADESI-MS/MS method was used to evaluate the chemical composition of HPE and the 2,2-diphenyl-1-picrylhydrazyl (DPPH) method was used to measure the antioxidant activity of geopropolis. Results: While HPE alone (G2) caused no liver injury, the livers of G4 exhibited color and histological alterations, extensive hemorrhagic suffusions, friable consistence, yellowish-white multifocal areas, degeneration and diffuse hepatocyte necrosis, in addition to altered levels of urea and enzymes TGO (glutamic-oxaloacetic transaminase) and TGP (glutamic pyruvic transaminase). HPE was shown to exert hepatoprotective effect in G3 animals, only mild hemorrhage and focal areas of vacuolar degeneration and hepatocyte necrosis having been observed. The geopropolis analyzed contains a high diversity of phenolic compounds, including chalcones, flavones and flavonols. high antioxidant activity was determined (IC50=48 μg/mL). Conclusions: The hydroethanolic extract of the geopropolis produced by M. subnitida has a high diversity of phenolic compounds and a high antioxidant activity. It has hepatoprotective action, as evidenced by a reduction in the levels of enzymaraes and the severity of the macroscopic and histological changes in livers of Wistar rats subjected to acetaminophen induced liver changes.

Clinical and histological features of patients with chronic hepatitis B virus infection in the grey zone.

Chronic HBV infection patients who do not conform to any of the usual immune states are regarded as 'grey zone' patients. We aimed to investigate the proportion of chronic HBV infection patients in the grey zone, and evaluate the clinical characteristics and liver pathological changes in grey zone patients. Clinical data of 1391 treatment-naive chronic HBV infection patients with liver biopsy were collected. Natural history of HBV infection was determined based on European Association for the Study of the Liver (EASL) 2017, American Association for the Study of Liver Diseases (AASLD) 2018 and Chinese 2019 guidelines for the prevention and treatment of chronic HBV infection. Significant liver histological changes and associated risk factors of normal ALT grey zone patients were analysed. According to EASL, AASLD and Chinese criteria, there were 50.0%, 28% and 37.4% chronic HBV infection patients in the grey zone. Among the 353 grey zone patients with normal ALT, 72.4% had significant liver histological changes. ALT (optimal cut-off value 25 IU/L) and HBV DNA (optimal cut-off value 18,000 IU/mL) were independent risk factors of significant liver histological abnormalities. In conclusion, a substantial proportion of grey zone patients with normal ALT have significant liver histological changes that can be predicted by levels of serum ALT and HBV DNA. These results provide guidance of antiviral treatment in grey zone patients.

Pharmacological effect of Argyrolobium roseum (Camb.) Jaub & Spach extracts against lead-induced toxicity in rats.

Argyrolobium roseum (Camb.) Jaub & Spach (Papilionaceae) is a medicinal plant, cultivated in northern areas of Pakistan. The consumption of trace minerals (lead) is very toxic to the vital organs of the body, therefore the overcome of these minerals is very necessary. In this regard, this study aimed to assess the potential pharmacological effect of aqueous and ethanolic extract of Argyrolobium roseum (Camb.) Jaub & Spach against pb-induced oxidative stress, histological changes in Pb-induced rats' liver and kidney, and anti-inflammatory effect. The metal concentrations in liver and kidney homogenates were measured through atomic absorption spectrophotometer. The antioxidant activity was measured through DPPH and FRAP assay. Pb concentrations were significantly higher in liver and kidney homogenates after injection of Pb acetate was given intraperitoneally (45.2 ± 6.8 and 58.8 ± 7.9, respectively; p < .0001). The level of Pb in liver and kidney homogenates was significantly reduced by aqueous and ethanolic extracts of Argyrolobium roseum (Camb.) Jaub & Spach. The Pb + Aq-600 mg/kg-treated rats exhibited a protective effect on hepatocytes cells against Pb-induced liver injury and restored the cells of the kidney. Pb + Aq-600 mg/kg showed higher antioxidant activity as compared to other treated groups. The highest decreased MDA level was found in liver and kidney homogenate of Pb + Aq-600 mg/kg rats (11.2 ± 1.51 nmol/mg; p < .001) and GSH and CAT levels tended to normal after treatment of Pb + Aq-600 mg/kg in rats. The ALAD, ALT, AST, and ALP level were enhanced and tended to be normal after the Aq-400 and Aq-600 mg/kg treatment in Pb-exposed rats. The result showed that 600 mg/kg Aq + Pb exhibited significant (p < .001) anti-inflammatory activity. The findings of this study concluded that treatment of the aqueous extract of Argyrolobium roseum (Camb.) Jaub & Spach reduces the renal and hepatic damage in Pb-induced rats and it also decreases oxidative stress via improving antioxidant components.

Naringin and naringenin counteract taxol-induced liver injury in Wistar rats via suppression of oxidative stress, apoptosis and inflammation

This research aimed to evaluate the preventing effects of naringin, naringenin, and their combination on liver injury induced by Taxol (paclitaxel) in Wistar rats. Male Wistar rats received 2 mg/kg Taxol intraperitoneal injections twice weekly on the second and fifth days of each week for 6 weeks. During the same period as Taxol administration, rats were given naringin, naringenin, or a combination of the two (10 mg/kg b.wt) every other day. Treatment with naringin and/or naringenin reduced the abnormally high serum levels of total bilirubin, aspartate transaminase, alanine transaminase, alkaline phosphatase, lactate dehydrogenase, and gamma-glutamyl transferase in Taxol-treated rats. It also significantly increased the level of serum albumin, indicating an improvement in the liver. The perturbed histological liver changes were markedly improved due to the naringin and/or naringenin treatment in Taxol-administered rats. Additionally, the treatments reduced high hepatic lipid peroxidation and increased liver glutathione content as well as the activities of superoxide dismutase and glutathione peroxidase. Furthermore, the treatments reduced the levels of alpha-fetoprotein and caspase-3, a pro-apoptotic mediator. The naringin and naringenin mixture appeared more effective in improving organ function and structural integrity. In conclusion, naringin and naringenin are suggested to employ their hepatoprotective benefits via boosting the body’s antioxidant defense system, reducing inflammation, and suppressing apoptosis.Graphical

Exposure of zebrafish (Danio rerio) to graphene oxide for 6 months suppressed NOD-like receptor-regulated anti-virus signaling pathways.

Environmental exposure to graphene oxide (GO) is likely to happen due to the use and disposal of these materials. Although GO-induced ecological toxicity has been evaluated before by using aquatic models such as zebrafish, previous studies typically focused on the short-term toxicity, whereas this study aimed to investigate the long-term toxicity. To this end, we exposed zebrafish to GO for 6 months, and used RNA-sequencing to reveal the changes of signaling pathways. While GO exposure showed no significant effects on locomotor activities, it induced histological changes in livers. RNA-sequencing data showed that GO altered gene expression profiles, resulting in 82 up-regulated and 275 down-regulated genes, respectively. Through the analysis of gene ontology terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, we found that GO suppressed the signaling pathways related with immune systems. We further verified that GO exposure suppressed the expression of genes involved in anti-virus responses possibly through the inhibition of genes involved in NOD-like receptor signaling pathway. Furthermore, NOD-like receptor-regulated lipid genes were also inhibited, which may consequently lead to decreased lipid staining in fish muscles. We concluded that 6 month-exposure to GO suppressed NOD-like receptor-regulated anti-virus signaling pathways in zebrafish.

The histopathological changes of liver and testis of Japanese quail chicks fed different levels of dietary L-valine

This experiment was carried out to investigate the histological changes of liver and testis of Japanese quail fed different levels of dietary valine (Val) in low protein diet. A total of 1000 one-day-old Japanese quail chicks (mixed sex) were assigned to five experimental diets including diets containing 7.5, 8.5, 9.5, 10.5 and 11.5 g digestible (dig.) Val/kg diet in a completely randomized design, with 5 replicates of 40 quail chicks per pen. Experimental diets were formulated to be isoenergetic and isonitrogenous (170 g crude protein/kg) to meet nutrients recommendation of growing quails suggested by Brazilian tables. At d 42, quail chicks were slaughtered, and tissue samples were collected and fixed to evaluate the histological indices of liver and testis. High levels of Val, increased (P < 0.05) diameter of liver cell nucleus and liver hepatocytes in both male and female. While 11.5 g Val showed mild hepatosteatosis, bile duct hyperplasia was observed in 10.5 g Val. In 7.5 and 8.5 g Val groups, there was no negative effects on the liver histology. The male quail chicks which fed on diets containing 8.5 g Val had better significant (P < 0.05) reproductive indexes [Tubular differentiation (TDI) and spermatic index (SI)]. In conclusion, the use of high levels of Val (≥ 9.5 g dig. Val/kg diet) during d 0 – 42 of age can lead to histological damage in liver and testis of quail chicks.

HEPATOPROTECTIVE EFFECT OF SEA BUCKTHORN BERRY SEED OIL IN CYCLOPHOSPHAMIDE-INDUCED HEPATIC TOXICITY IN BALB/c MICE

Background: Chemotherapy-induced hepatotoxicity can be mitigated by use of antioxidants, which may help the liver to recover its endogenous antioxidant mechanism. The objective of this study was to determine the effectiveness of sea buckthorn berry seed oil (SBO) in attenuating cyclophosphamide-induced changes in liver enzymes and liver histology in BALB/c mice. Methods: This experimental study was conducted in Physiology Department of Foundation University School of Health Sciences (FUSH), in collaboration with Pathology Department, FUSH and National Institute of Health, Islamabad from Jan 2018 to Jun 2019. Thirty healthy male BALB/c mice were divided into 3 groups of 10 each. Group-1 served as control. Group-2 received cyclophosphamide (25 mg/Kg body weight) intraperitoneally for 10 days. Group-3 was co-administered cyclophosphamide (same dose) with of sea buckthorn berry seed oil (40 mg/Kg body weight) orally for ten days. All animals were sacrificed on 11th day. Serum levels of liver enzymes as liver injury biomarkers were assayed. Liver histopathology was carried out for evidence of hepatic injury and recovery. Results: Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP) increased significantly in Group-2 (p&lt;0.05). In Group-3, the rise in hepatic injury serum markers was significantly less than Group-2 (p&lt;0.05). Upon histological examination, Group-2 was grade 5 according to Kleiner’s scoring system for steatohepatitis, and had grade 2 sinusoidal dilatation on Rubbia-Brandt grading system. These changes were significantly less in Group-3 (p&lt;0.05). Conclusion: Co-administration of SBO mitigated the CP-induced rise in hepatic injury biomarkers and sinusoidal injury. Pak J Physiol 2023;19(2):20–4

Protective role of Dodonaea viscosa extract against streptozotocin-induced hepatotoxicity and nephrotoxicity in rats

Previous investigations have shown that D. viscosa herbal extract is often used to treat a variety of diseases. Therefore, the purpose of this study was to investigate any additional potential impacts on rat liver and kidney damage induced by diabetes. Streptozotocin (STZ) (60 mg/kg/day) was given as a single dosage to cause type 1 diabetes. After then, diabetic rats received oral doses of D. viscosa for four weeks at 150 and 300 mg/kg/day. Blood, liver, and kidney tissues were collected at the end of the treatment and examined. Analysis was made of the serum lipid profile, liver, and kidney functions, as well as blood biochemistry. Moreover, the levels of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), interleukin-1 beta (IL-1β), prostaglandin E-2 (PGE-2), and nitric oxide (NO) were estimated in serum. In liver and kidney samples, thiobarbituric acid reactive substances (TBARs) and reduced glutathione (GSH), as well as the pro-inflammatory cytokines and enzymatic activities of glutathione peroxidase (GPx), glutathione reeducates (GR), glutathione-S-transferase (GST), catalase (CAT), and superoxide dismutase (SOD) were analyzed. Histological changes in liver and kidney cross-sections were also observed. Our findings demonstrated that D. viscosa dramatically decreased pro-inflammatory indicators in blood, kidney, and liver tissues as well as blood glucose, and restored insulin levels, and lipid profiles. Additionally, it significantly raises the antioxidant enzyme activity SOD, CAT, GPx, and GST, while significantly lowering TBARs levels. The above-mentioned biochemical changes that took place in tissues were further supported by histological alterations. These findings imply that D. viscosa protects against STZ-induced hyperglycemia, aberrant lipid synthesis, and oxidative stress and that these benefits may be mediated by interacting with various targets to increase the levels of antioxidant enzymes in the liver and kidneys. Its mode of action and safety for use as medicine against various metabolic problems caused by diabetes require more research.

Histological changes in liver and kidney of male mice by age after exposure to aluminum chloride.

The informal aluminum industry is increasingly widespread in low- and middle-income countries, especially in Indonesia. Aluminum exposure is a serious public health problem, especially among workers in the informal aluminum foundry sector. Research on aluminum (Al) is important to advance our understanding of its impact on physiological systems. Here, we investigated the effect of exposure to aluminum longitudinal histological changes on the liver and kidneys of male mice. Mice were separated into six groups (4/group): group 1, group 2, group 3 received vehicles, and group 4, group 5, group 6 were administered a single dose of Al at 200mg/kg b.w. by intraperitoneally every 3days for 4weeks. Post-sacrifice, kidneys and liver were isolated for examination. While Al did not impact the body weight gain of male mice across all groups, it caused liver damage including sinusoidal dilatation, enlarged central veins, vacuolar degeneration, and pyknotic nuclei in one-month-old mice. Furthermore, atrophied glomeruli, blood-filled spaces, and disintegration of renal tubular epithelium are evident at one-month-age. By contrast, sinusoidal dilatation and enlarged central veins were found in mice two- and three-months-old, including hemorrhage in mice (two-month-old) and atrophy of glomeruli. Lastly, the kidneys of three-month-old mice displayed interstitial fibrosis and increasing mesenchyme in the glomeruli. In summary, we demonstrated that Al provoked histological changes in the liver and kidneys with Al-treated 1-month mice being the most susceptible.

A polysaccharide from Codonopsis pilosula roots attenuates carbon tetrachloride-induced liver fibrosis via modulation of TLR4/NF-κB and TGF-β1/Smad3 signaling pathway.

The present work reported the extraction, purification, characterization of a polysaccharide from roots of Codonopsis pilosula (CPP-A-1) and its effect on liver fibrosis. The findings exhibited that the molecular weight of CPP-A-1 was 9424Da, and monosaccharide composition were glucose and fructose and minor contents of arabinose. Structural characterization of CPP-A-1 has a backbone consisting of→(2-β-D-Fruf-1)n→ (n ≈ 46-47). Treatment with CPP-A-1 inhibited the proliferation of transforming growth factor-beta 1 (TGF-β)-activated human hepatic stellate cell line (LX-2), and induced cell apoptosis. We used carbon tetrachloride (CCl4) to construct mice model of liver fibrosis and subsequently administered CPP-A-1 treatment. The results showed that CPP-A-1 alleviated CCl4-induced liver fibrosis as demonstrated by reversing liver histological changes, decreased serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) contents, collagen deposition, and downregulated fibrosis-related collagen I and α-smooth muscle actin (α-SMA), and inhibited the generation of excessive extracellular matrix (ECM) components by restoring the balance between matrix metalloproteinases (MMPs) and its inhibitor (TIMPs). Moreover, CPP-A-1 improved anti-oxidation effects detected by promoting liver superoxide dismutase (SOD), glutathione (GSH) and Mn-SOD levels, and inhibition of liver malondialdehyde (MDA) and iNOS levels. CPP-A-1 also ameliorated the inflammatory factor (tumor necrosis factor-alpha (TNF-α) and interleukin (IL)-6), and expression of inflammatory factor genes (TNF-α, IL-11 mRNA). In addition, our results showed that CPP-A-1 inhibited Toll-like receptor 4 (TLR4)/nuclear factor kappa-B (NF-κB) and transforming growth factor-β1 (TGF-β1)/drosophila mothers against decapentaplegic 3 (Smad3) signaling pathways. Furthermore, In vitro tests of LX-2 cells demonstrated that CPP-A-1 not only inhibited α-SMA expression with lipopolysaccharide (LPS) or TGF-β1 stimulation, but also inhibited TLR4/NF-κB and TGF-β1/Smad3 signaling, similar to corresponding small-molecule inhibitors. Therefore, CPP-A-1 might exert suppressive effects against liver fibrosis by regulating TLR4/NF-κB and TGF-β1/Smad3 signaling, our findings support a possible application of CPP-A-1 for the treatment of liver fibrosis.

Efficient prime editing in mouse brain, liver and heart with dual AAVs

Realizing the promise of prime editing for the study and treatment of genetic disorders requires efficient methods for delivering prime editors (PEs) in vivo. Here we describe the identification of bottlenecks limiting adeno-associated virus (AAV)-mediated prime editing in vivo and the development of AAV-PE vectors with increased PE expression, prime editing guide RNA stability and modulation of DNA repair. The resulting dual-AAV systems, v1em and v3em PE-AAV, enable therapeutically relevant prime editing in mouse brain (up to 42% efficiency in cortex), liver (up to 46%) and heart (up to 11%). We apply these systems to install putative protective mutations in vivo for Alzheimer’s disease in astrocytes and for coronary artery disease in hepatocytes. In vivo prime editing with v3em PE-AAV caused no detectable off-target effects or significant changes in liver enzymes or histology. Optimized PE-AAV systems support the highest unenriched levels of in vivo prime editing reported to date, facilitating the study and potential treatment of diseases with a genetic component.

Phytochemical profiling, antioxidant potential and protective effect of leaves extract of tunisian Vitis vinifera autochthonous accessions against acute CCl4-injured hepatotoxicity in mice

Vitis vinifera leaves (VVL) are agro-industrial waste. In the current study, the phytochemical profile of V. vinifera leaves extracts (VVLE) of two Tunisian autochthonous accessions was determined via LC-UV-ESI/MS, and their antioxidant and hepatoprotective properties were also assessed. Mice were pretreated orally with VVLE (7.5, 15 and 30 mg/kg) for 7 days, and then received acutely and by i.p. a solution CCl4 at 12% in sunflower oil (v/v). Serum levels of hepatic markers, oxidative stress indicators in liver tissue and histological changes were assessed. LC-UV-ESI/MS analysis revealed four phenolic compounds identified in both extracts with quercetin-3-O-glucuronide being the dominant constituent (23.32 ± 1.06 vs. 10.24 ± 0.12 mg/g DM, p < 0.05 for wild and cultivated accessions, respectively). The Antioxidant activity revealed a significant difference between the genotypes. Moreover, the VVLE of the wild "Nefza-I" ecotype was the most active based on antioxidant assays. Furthermore, the results showed that pre-treatment, especially with VVLE, of the wild ecotype "Nefza-I", attenuated CCl4-induced acute liver injury in a dose-dependent manner, as demonstrated by the decrease in the activities of hepatic serum function markers. This was also evidenced by a decrease in the levels of lipoperoxidation and histological damage in the liver, as well as a restoration of antioxidant enzyme activities (SOD and catalase) and an increase in the hepatic glutathione content. Our results demonstrate that VVLE possesses protective effects on CCl4-induced liver injury. Overall, the wild ecotype "Nefza-I" extract could serve as an effective protector against CCl4-induced hepatocellular oxidative stress.

Detection and Quantification of Some Ethanol-Producing Bacterial Strains in the Gut of Mouse Model of Non-Alcoholic Fatty Liver Disease: Role of Metformin.

Ethanol-producing dysbiotic gut microbiota could accelerate the progress of non-alcoholic fatty liver disease (NAFLD). Metformin demonstrated some benefits in NAFLD. In the present study, we tested the ability of metformin to modify ethanol-producing gut bacterial strains and, consequently, retard the progress of NAFLD. This 12-week study included forty mice divided into four groups (n = 10); normal diet, Western diet, Western diet with intraperitoneal metformin, and Western diet with oral metformin. Oral metformin has a slight advantage over intraperitoneal metformin in ameliorating the Western diet-induced changes in liver function tests and serum levels of different cytokines (IL-1β, IL-6, IL-17, and TNF-α). Changes in liver histology, fibrosis, lipid content, Ki67, and TNF-α were all corrected as well. Faecal ethanol contents were increased by the Western diet but did not improve after treatment with metformin although the numbers of ethanol-producing Klebsiella pneumoniae (K. pneumoniae) and Escherichia coli (E. coli) were decreased by oral metformin. Metformin did not affect bacterial ethanol production. It does not seem that modification of ethanol-producing K. pneumoniae and E. coli bacterial strains by metformin could have a significant impact on the therapeutic potentials of metformin in this experimental model of NAFLD.

Potential therapeutic action of tauroursodeoxycholic acid against cholestatic liver injury via hepatic Fxr/Nrf2 and CHOP-DR5-caspase-8 pathway.

Cholestasis is a pathophysiologic syndrome with limited therapeutic options. Tauroursodeoxycholic acid (TUDCA) has been employed to treat hepatobiliary disorders and is as effective as UDCA in alleviating cholestatic liver disease in clinical trials. Until now, TUDCA's mechanism of action toward cholestasis remains unclear. In the present study, cholestasis was induced with a cholic acid (CA)-supplemented diet or α-naphthyl isothiocyanate (ANIT) gavage in wild-type and Farnesoid X Receptor (FXR) deficient mice, using obeticholic acid (OCA) as control. The effects of TUDCA on liver histological changes, transaminase level, bile acid composition, hepatocyte death, expression of Fxr and nuclear factor erythroid 2-related factor 2 (Nrf2) and target genes, as well as apoptotic signaling pathways, were investigated. Treating CA-fed mice with TUDCA markedly alleviated liver injury, attenuated bile acids retention in liver and plasma, increased Fxr and Nrf2 nuclear levels and modulated the expression of targets regulating synthesis and transportation of bile acids, including BSEP, MRP2, NTCP and CYP7A1. TUDCA, but not OCA, activated Nrf2 signaling and exerted protective effects against cholestatic liver injury in Fxr-/- mice fed with CA. Furthermore, in both mice with CA- and ANIT-induced cholestasis, TUDCA decreased expression of GRP78 and CCAAT/enhancer-binding protein homologous protein (CHOP), reduced death receptor 5 (DR5) transcription, caspase-8 activation, and BID cleavage, and subsequently inhibited activation of executioner caspases and apoptosis in liver. We confirmed that TUDCA protected against cholestatic liver injury by alleviating BAs burden of dually activating hepatic Fxr and Nrf2. Moreover, inhibiting CHOP-DR5-caspase-8 pathway contributed to the anti-apoptotic effect of TUDCA in cholestasis.