Liver Histopathological Features Research Articles

Multi-target regulatory effects of rhaponticin in a rat model of hepatic fibrosis revealed by non-targeted metabolomics

IntroductionHepatic fibrosis (HF), a progressive chronic liver disease, is a serious threat to global public health. The lack of preventive and therapeutic strategies has created an urgent need for effective anti-fibrosis agents. There is growing evidence that natural products might provide safe and effective interventions for HF. Among them, rhaponticin (RHA), a stilbenoid glucoside natural product isolated from medicinal plants of Rheum L. of Polygonaceae Juss. has many pharmacological activities such as anti-inflammatory, antioxidant, antiproliferative, and antithrombotic properties. However, its effects on HF remain unclear.MethodsHerein, we investigated the effects of RHA against HF on the carbon tetrachloride (CCl4)-induced hepatic fibrosis and the underlying mechanism in rats. Functional, histopathological, and protein-level indicators of liver insult were evaluated. Moreover, serum metabolites were assessed by non-targeted metabolomics.Results and discussionThe results showed that RHA improved liver functions and histopathological features in the liver of CCl4-treated rats, and alleviated the expression of α-SMA and type I collagen. Meanwhile, RHA also modulated endogenous metabolite levels in rats with HF, targeting glycerophospholipid metabolism signaling and other pathways. These findings confirmed the protective effects of RHA against hepatic fibrosis in rats by exerting multi-target effects via multiple signaling and metabolic pathways. Which may be of use in developing more effective RHA-based therapeutic strategies for hepatic fibrosis.

Desloratadine mitigates hepatocellular carcinoma in rats: Possible contribution of TLR4/MYD88/NF-κB pathway.

Chemotherapeutic medication-induced systemic toxicity makes cancer treatment less effective. Thus, the need for drug repurposing, which aids in the development of safe and efficient cancer therapies, is urgent. The primary goal of this research was to assess desloratadine hepatoprotective abilities and its capacity to attenuate TLR4/MyD88/NF-κB inflammatory pathway in hepatocellular carcinoma (HCC) induced by thioacetamide (TAA). Male Sprague Dawely rats received TAA injections (200mg/kg, i.p., 2 times/week) for 16weeks. To confirm the development of HCC, liver function biomarkers and histopathological analysis were evaluated. Desloratadine (5mg/kg, p.o.) was administered to rats in 2 treatment groups; HCC+DES 1 group received desloratadine with TAA for 1month from week 13-16, HCC+DES 2 group received desloratadine with TAA for 2months from week 9-16. Chronic TAA administration resulted in considerable overexpression of the profibrogenic cytokine TGF-β and elevation in protein expression of NF-κB besides levels of TLR4, MyD88, TRAF6, TAK1 and IL-1β. Desloratadine administration showed a significant improvement in liver function tests, as well as an increase in tissue antioxidant enzymes and an improvement in the liver's histopathological features. Collectively, desloratadine through modulating TLR4/MyD88/TRAF6/TAK1/NF-κB and acting as an antioxidant, is a promising treatment for HCC induced by TAA.

Kadukkai maathirai (Indian herbal drug) prevents hepatocellular cancer progression by enhancing GSTM1 expression and modulating β catenin transcription: in-silico and in-vivo study

Background Hepatocellular carcinoma (HCC) is an aggressive malignancy with poor clinical outcomes. Hence cost-effective drugs with fewer side effects as a standard supportive therapy might yield substantial advantages in efficacy and safety. Kadukkai maathirai (KM) is being used as a supplement in hepatocellular carcinoma. We evaluated whether KM has any preventive action on cancer progression in diethyl nitrosamine (DEN) - induced HCC in rats. Methods DEN was injected to produce HCC in rats, which was confirmed after 16 weeks. All the rats were orally administered KM for 4 weeks. Hepatoprotective potential (serum AST, ALT, ALP, Bilirubin) and anticancer efficacy (body weight, nodule count, tumor progression by histopathology, expression of GSTM1 by Liquid chromatography-mass spectrometry (LC-MS), and In-silico analysis of phytoconstituents against β catenin and LRP analysis were evaluated. Results KM prevented cancer progression against DEN-induced HCC by an increase in GSTM1, a phase II detoxifying enzyme. It significantly reversed altered nodule count, relative liver weight, body weight, and histopathological features of HCC. In silico analysis of phytoconstituents of KM showed that they modulate the intracellular transcription process by inhibiting the armadillo repeat region of β catenin. Conclusions Our results elucidate the potential of KM as a supplement in HCC by reducing nodule count, protecting the liver from further damage, GSTM1 expression, and inhibiting armadillo repeat region of β catenin.

Kadukkai maathirai (Indian herbal drug) prevents hepatocellular cancer progression by enhancing GSTM1 expression and modulating β catenin transcription: in-silico and in-vivo study

Background Hepatocellular carcinoma (HCC) is an aggressive malignancy with poor clinical outcomes. Hence cost-effective drugs with fewer side effects as a standard supportive therapy might yield substantial advantages in efficacy and safety. Kadukkai maathirai (KM) is being used as a supplement in hepatocellular carcinoma. We evaluated whether KM has any preventive action on cancer progression in diethyl nitrosamine (DEN) - induced HCC in rats. Methods DEN was injected to produce HCC in rats, which was confirmed after 16 weeks. All the rats were orally administered KM for 4 weeks. Hepatoprotective potential (serum AST, ALT, ALP, Bilirubin) and anticancer efficacy (body weight, nodule count, tumor progression by histopathology, expression of GSTM1 by Liquid chromatography-mass spectrometry (LC-MS), and In-silico analysis of phytoconstituents against β catenin and LRP analysis were evaluated. Results KM prevented cancer progression against DEN-induced HCC by an increase in GSTM1, a phase II detoxifying enzyme. It significantly reversed altered nodule count, relative liver weight, body weight, and histopathological features of HCC. In silico analysis of phytoconstituents of KM showed that they modulate the intracellular transcription process by inhibiting the armadillo repeat region of β catenin. Conclusions Our results elucidate the potential of KM as a supplement in HCC by reducing nodule count, protecting the liver from further damage, GSTM1 expression, and inhibiting armadillo repeat region of β catenin.

Protective effect of olive leaf (Olea europaea L.) extract against chronic exposure of liver and kidney tissues of Wistar rats to aluminum chloride

Introduction: The liver and kidney are the main sites of aluminum (Al) accumulation. Lifetime exposure to significant amounts of Al is inevitable, hence its toxicity on the liver and kidney should be a health concern. Natural antioxidants have been proven to alleviate pathologies in various liver and kidney injuries. However, the effect of olive leaf extract (OLE) on Al-exposed animals is yet to be confirmed. This study aimed to investigate the OLE effect against AlCl3 chronic exposure in rats’ liver and kidneys. Methods:Thirty-two male Wistar rats were divided into four groups (n=8), including the control group, the AlCl3 group treated with 128 mg/kg AlCl3 solution, as well as AlCl3 +OLE50, and AlCl3 +OLE100 groups (Other than AlCl3 they received 50 and 100 mg/kg of OLE, respectively, 2 hours after AlCl3 administration). All treatments were given orally for 12 weeks. All groups were evaluated for liver and kidney histopathological features, then scoring was performed. Results: AlCl3 administrations produced histopathological lesions in the liver and kidney, indicated by increased liver necro-inflammatory grades, ballooning scores, and renal inflammatory cell infiltration (P<0.05). OLE100 mg/kg significantly reduced liver necroinflammatory grade, ballooning score, and kidney inflammatory cell infiltrations. The dose of 50 mg/kg also reduced these parameters (P<0.05), except for the liver necro-inflammatory grade. There was a significant correlation between OLE dose and liver necro-inflammatory grade and ballooning score amelioration. Conclusion: OLE ameliorates liver and kidney histopathological features induced by oral Al chronic exposure in a dose-dependent manner.

The Effect of Giving Salak (Salacca Zalacca) Bark Extract on Liver Function and Histopathological Features of the Liver of Male Wistar White Rats Obesity Model

Nonalcoholic fatty liver disease (NAFLD) Lifestyle factors contribute to fatty liver disease. Physical activity, weight loss, and antioxidant-rich components like Salak herb are used to minimize liver disease. The study examined how Salak bark extract affected liver function in white male Wistar rats, an obesity model, and histopathology. An actual experiment or laboratory experimental design is used in this quantitative research. Four experimental groups of 24 mice were employed. Research data was analyzed with SPSS 25.0. Normality test findings reveal 2-tailed significance of 0.991 > 0.05. The One-Way ANOVA test at a 95% confidence level for the SGOT liver function analysis data showed the same variance, 0.233 (p>0.05) and 0.718 (p>0.05) for the SGPT liver function analysis. Histopathological studies showed snake fruit extract at 300mg/BW scored 3 (Hydropic degeneration), 500mg/BW scored 2 (Parenchymatous degeneration), and 700mg/BW scored 1 (Normal). The phytochemical screening of alkaloids, flavonoids, saponins, tannins, and glycosides demonstrated that Salak bark extract's secondary metabolite concentration contains antioxidant-effective active components. The study demonstrated no inflammation, cell improvement, necrosis, or fat at 700 mg/BW.

Effects of Moringa oleifera Leaf Extract on Liver Histopathology: A Systematic Review.

Moringa leaves (Moringa oleifera), which are members of the Moringaceae family, are one of the herbal plants that are widely known in Indonesia. Phytochemical contents of moringa leaf, such as flavonoid, quercetin, and phenolic acid, are believed to have an effect on improvement of NAFLD. Therefore, moringa leaf is considered as one the herbal plants that can be used as supplementation in the form of adjuvant therapy to NAFLD. The study objective of our research is to review the effect of giving moringa leaf to the liver, especially the histopathologic features. This study will be conducted on literature review research design, more specifically in the form of a systematic review. Research Method. Five major electronic web databases, including PubMed, Cochrane Library, Google Scholar, Scopus, and ScienceDirect, were used in identifying literature from 2014 to 2023. From a comprehensive analysis of 13 relevant literature sources, we elucidate the impact of Moringa oleifera leaf extract on liver histopathology, glucose, and lipid metabolism. Furthermore, we provide insights into its safety profile concerning human health. The phytochemical content of Moringa oleifera leaf extract had shown a significant benefit in plant medicinal sector. From the research that had been done, Moringa oleifera leaf extract contributes to give significant improvement on liver histopathological features, glucose, and lipid metabolism on animal sample model.

Noninvasive Tests to Assess Liver Stiffness in Patients with Chronic Hepatitis B: APRI, FIB‐4, and FIB‐5 Scores

Background. Invasive percutaneous liver parenchymal biopsy is the best test used to evaluate liver stiffness and fibrosis in the follow‐up and treatment of chronic hepatitis B (CHB) patients. In this study, we aimed to indirectly evaluate the severity of liver parenchymal fibrosis with tests used in the laboratory. Methods. This retrospective study was conducted with 201 patients diagnosed with CHB who underwent liver biopsy between 2021 and 2022. Preprocedural examination information, laboratory tests, and histopathological data of the patients were taken from the hospital database and examined. “Aspartate aminotransferase (AST)‐platelet ratio index” (APRI), “4 factor‐based fibrosis index” (FIB‐4) score, and “5 factor‐based fibrosis index” (FIB‐5) score were calculated and compared with liver histopathological features. Results. Of the 201 CHB patients, 76 were females and 125 were males. The average age of the patients was 38.05 ± 12.63 years. A weak, statistically significant correlation was observed between FIB‐4 and APRI scores. The patients’ significant fibrosis scores were 31.3% and 33.8%, respectively (r = 0.313; r = 0.338; p = 0.001; p < 0.01). The very weak negative correlation of 17.4% between the patients’ FIB‐5 score and fibrosis score was statistically significant (r = −0.174; p = 0.014; p < 0.05). Conclusions. According to the data we obtained in our study, while the APRI score and FIB‐4 score can be used safely, more comprehensive studies are needed for the reliability of the FIB‐5 score.

High Fischer ratio oligopeptides from hard-shelled mussel: Preparation and hepatoprotective effect against acetaminophen-induced liver injury in mice

High Fischer ratio oligopeptides (HFOPs) have a variety of physiological activities, but there are few literatures reporting the hepatoprotective effects of HFOPs against acetaminophen (APAP)-induced liver injury. The objectives of this study were to establish the preparation process of HFOPs from hard-shelled mussel (Mytilus coruscus) (HFOPs-HM) and investigate the hepatoprotective effects and potential mechanisms of HFOPs-HM against APAP-induced liver injury in mice. Therefore, trypsin and papain were selected as the endo and exogenous proteases for preparing HFOPs-HM according to the degree of hydrolysis (DH) of hard-shelled mussel hydrolysates, and activated carbon of XHJ-200 displayed the best effect to remove aromatic amino acids according to the Fischer ratio (OD220/OD280) and its parameters were optimized as adsorption time of 3.0 h and solid-liquid ratio of 1:10. The HFOPs-HM's Fischer ratio and average molecular weight were 26.03 (>20) and 878.54 Da, respectively. Furthermore, a mice model of liver injury induced by APAP was established. HFOPs-HM could significantly improve liver index and histopathological features, decrease the serum alanine aminotransferase (ALT) and aspartate transaminase (AST) activities, increase the glutathione content and superoxide dismutase (SOD) and catalase (CAT) activities. Further research demonstrated that HFOPs-HM could alleviate liver oxidative stress in liver tissue via activating the Nrf2 pathway to initiate the intracellular antioxidant enzyme system. In addition, HFOPs-HM also suppressed liver inflammation by downregulating the secretion of TNF-α, IL-1β, and IL-6. Thus, HFOPs-HM could serve as an auxiliary functional dietary molecule applied in function products to alleviate drug-induced liver injury.

Applicability of International Autoimmune Hepatitis Group (IAIHG) Scoring System for Autoimmune Hepatitis in Pediatrics.

Many hepatologic pathologies mimic autoimmune hepatitis (AIH). Researchers developed the International Autoimmune Hepatitis Group (IAIHG) scoring system to compensate for the lack of specific diagnostic tests for AIH. The scoring system was not designed with pediatric patients in mind, so there are limits to its pediatric use. Additionally, there is limited information on the value of a liver biopsy in conjunction with its use. In this retrospective study, we evaluated the effect of liver biopsy scores on the IAIHG scoring system in patients that were 0-18 years old with suspected AIH. We also analyzed demographic data and laboratory values associated with a final AIH diagnosis. We found that interface hepatitis and predominant plasma cells found during the biopsy were significantly associated with a final AIH diagnosis. We also found that abnormal laboratory values were associated with an AIH diagnosis. We found that IAIHG scores calculated post-liver biopsy showed a greater area under the receiver operating characteristic curve (AUROC) of 0.95, which was compared to 0.88 for the scores calculated before a liver biopsy. Including biopsy metrics lowered the optimized cutoff score and test specificity. Incorporating liver histopathological features improved the performance of the IAIHG scoring system. Further studies to identify other potential elements in liver histology may improve the performance metrics of the IAIHG test in the pediatric population.

Sea Cucumber Saponins Derivatives Alleviate Hepatic Lipid Accumulation Effectively in Fatty Acids-Induced HepG2 Cells and Orotic Acid-Induced Rats.

The sulfated echinoside A (EA) and holothurin A (HA) are two prominent saponins in sea cucumber with high hemolytic activity but also superior lipid-lowering activity. Deglycosylated derivatives EA2 and HA2 exhibit low hemolysis compared to EA and HA, but their efficacies on lipid metabolism regulation remains unknown. In this study, fatty acids-treated HepG2 cells and orotic acid-treated rats were used to investigate the lipid-lowering effects of sea cucumber saponin derivatives. Both the saponin and derivatives could effectively alleviate lipid accumulation in HepG2 model, especially EA and EA2. Moreover, though the lipid-lowering effect of EA2 was not equal with EA at the same dosage of 0.05% in diet, 0.15% dosage of EA2 significantly reduced hepatic steatosis rate, liver TC and TG contents by 76%, 41.5%, and 63.7%, respectively, compared to control and reversed liver histopathological features to normal degree according to H&E stained sections. Possible mechanisms mainly included enhancement of fatty acids β-oxidation and cholesterol catabolism through bile acids synthesis and excretion, suppression of lipogenesis and cholesterol uptake. It revealed that the efficacy of EA2 on lipid metabolism regulation was dose-dependent, and 0.15% dosage of EA2 possessed better efficacy with lower toxicity compared to 0.05% dosage of EA.

Effect of Garlic Extract on Liver Histopathology of BALB/c Mice with Nicotine Exposure

Nicotine affects liver in the presence of free radicals. One of the antioxidants is allicin which is the most abundant organosulfur in garlic. The objective of this study is to determine the effect of garlic extract as an antioxidant on the liver histopathological features of BALB/c mice exposed to nicotine. This is an experimental study with with post-test only control group design. The research subject were 24 male BALB/c mice. The K- group was the healthy control. The K+ group was exposed to inhaled nicotine of 10 mg/kgBW. Groups P1 and P2 were exposed to 10 mg/kgBW inhaled nicotine and were given garlic extract 300 mg/kgBW and 500 mg/kgBW. Treatment was given for 14 days. Data were analyzed by Kruskal-Wallis test and considered significant if the p value 0.05 with 95% confidence interval. Liver degeneration was grouped into normal, parenchymatous degeneration, hydropic degeneration, and necrosis. The results in the following groups: K: 80%, 20%, 0%, 0%; K(-) :0%,0%,87%,13% ; P1 : 0%,60%,40%,0%; P2 : 0%,100%,0%,0%. There was a significant difference in the degree of degeneration between groups (p=0.001). Garlic extract can reduce the degree of microscopic damage to the liver of BALB/c mice exposed to nicotine inhalation.

Grape (Vitis vinifera L.) skin extract reduced levels of SGPT and SGOT and improved the liver tissue structure of Wistar rats (Rattus novergicus) fed a high-cholesterol diet

Background: This study determined the effect of adding grape skin extract (Vitis vinifera L.) to the diet of Wistar rats fed with high cholesterol on levels of SGPT and SGOT and the structure of liver tissue. Methods: This study used 5 groups, namely control (standard diet), P1 (high cholesterol diet), P2 (high cholesterol diet and extract of grape skin as much as 100 mg/200 gBW/day), P3 (high cholesterol diet and extract of grape skin as much as 250 mg/200 gBW/day) and P4 (high cholesterol diet and extract of grape skin 500 mg/200 gBW/day). The study began with the production of extracts, the extracts dosage determination, high cholesterol diet preparation, sample (animals) preparation, extract addition to animals, SGPT and SGOT levels measurement, rat liver histopathology preparation and histopathological features observation, then ended with data analysis. Results: The results showed that administration of grape skin extract in Wistar rats fed a high-cholesterol diet affected the levels of SGPT and SGOT, as well as parenchymatous degeneration of hepatocytes. Administration of grape skin extract as much as 500 mg/200 g BW/day had the lowest mean levels of SGPT and SGOT and the lowest parenchymatous degeneration of hepatocytes. Conclusion: Thus, the extract of grape skin (Vitis vinifera L.) has the potential of hepatoprotector because it contains anthocyanin, which functions as an antioxidant and protects against liver damage.

Effects of garlic and its major bioactive components on non-alcoholic fatty liver disease: A systematic review and meta-analysis of animal studies

• Garlic had beneficial effects on the liver's histopathological features, fat content, enzymes, and weight. • Types and duration of interventions, disease and study models, and risk of bias did not affect the pooled results. • Fermented and raw garlic had similar effects on liver enzymes and weight. • The certainty of the evidence was very low to low according to GRADE. This systematic review and meta-analysis study was conducted to summarize the effects of garlic or its major components on hepatic triglyceride and cholesterol content, serum alanine transaminase (ALT) and aspartate transaminase (AST) levels, and liver weight. We searched PubMed, Embase, Scopus, and Web of Knowledge databases. Data were pooled, and standardized mean difference (SMD) and 95% confidence intervals (CI) were estimated using the random-effect model. Out of 958 reports, 28 articles were eligible, of which 22 studies were included in the meta-analysis. Most of the included studies demonstrated the beneficial effects of garlic on hepatic histopathological features. The pooled results showed that garlic significantly decreased hepatic triglyceride and cholesterol, ALT, AST, and liver weight. The certainty of the estimates was very low to low according to GRADE. In conclusion, our study demonstrated amelioration of hepatic histopathological characteristics, hepatic lipid content, serum liver enzymes, and liver weight following garlic administration.

Effect of High Fat Diet on Histopathological Appearance of Pregnant Wistar Rat’s Liver

Indonesia is facing increased consumption of calorie-dense foods, such as fast food, among its society. Changes in diet and physical activities towards unhealthy ones can result in an imbalanced lifestyle, leading to various consequences, including hyperlipidemia. Pregnant women experience significant increases in plasma concentrations of lipids, lipoproteins, and apolipoproteins. Excessive cholesterol levels in pregnant women will result in the formation of fat in the liver. The purpose of this study was to determine the effect of a high-fat diet to pregnant Wistar rats (Rattus norvegicus) on the histopathological features of the liver. A true experimental with post test only control group design was applied in this study, which was conducted from August 2020 to November 2021. Fourteen (14) pregnant female rats were divided into two groups. The first group of rats was fed with a high-fat diet, while the second group, which was the control group, was given a standard diet. The high-fat diet used was 3 mL/rat/day beef brain. After the rats delivered their litters, the rats’ livers were retrieved, then examined histopathologically under a microscope. This research was conducted from No fatty liver was identified in the control group or the treatment group. Hence, a high-fat diet does not affect the liver histopathological features in pregnant Wistar rats.

Effect of Phylantus ninuri leaf extract on kidney and liver histopathological features in formaldehyde-exposed rats

Formaldehyde (FA) metabolism in the body reduces glutathione levels, resulting in liver and kidney damage. Phylantus ninuri (meniran) has been demontrated to have hepatoprotective properties. However, the effect of meniran administration on the histological features of the liver and kidney in in formaldehyde-exposed rats has not been adequately studied. The study was aimed to determine the effect of meniran on the histopathological feature of liver and kidney cell. This was a posttest randomized control group design. A total of 24 male Wistar rats were divided into 4 groups of 6 rats each. Group 1 was a control group that received only oral formaldehyde. Group 2, Group 3, Group 4 was administered oral formaldehyde together with meniran extract at dose of 100 mg/kg BW, 200 mg/kg BW, and 300 mg /kg BW respectively for 14 days. The histopathological changes of the kidneys and liver were assesed using hematoxylin and eosin (HE) staining. The mean of kidney and liver damage in the meniran group at various doses was lower than that of control group. There was a significant difference in the mean of kidney damage between the control group and the meniran group at various doses (p<0.05). The mean histopathological score of liver damage showed a significant difference between the control group and various treatments (p<0.05). Ethanolic extract of meniran leaves ameliorates liver and kidney damage in formaldehyde-exposed rats.

Histological growth patterns of colorectal cancer liver metastases: a strong prognostic marker associated with invasive patterns of the primary tumor and p53 alteration

The histological growth pattern of liver metastases (desmoplastic, pushing, and replacement patterns) at the tumor-liver parenchymal interface is a prognostic factor in patients with colorectal cancer. However, data regarding its association with the primary tumor characteristics and molecular alterations are limited. This study evaluated the histological growth pattern in 136 cases of colorectal cancer liver metastases without preoperative treatment, comparing it with the clinicopathological features of the primary tumor. Liver metastasis exhibiting predominantly non-desmoplastic pattern (<50%), observed in 74 cases (54%), was associated with hepatic vein invasion (P=0.025), worse recurrence-free survival (P<0.001) and overall survival (P=0.008). In multivariate analyses, multiple tumors (P<0.001) and non-desmoplastic patterns (P=0.009) were associated with worse recurrence-free survival, and tumor size (P=0.025) and non-desmoplastic pattern (P=0.025) were associated with worse overall survival. In 88 patients with available primary tumor tissue slides, non-desmoplastic pattern in the liver metastasis was associated with high-grade tumor budding (P=0.002), high-grade poorly differentiated cluster (P=0.021), absence of mucinous histology (P=0.016), and aberrant p53 expression (complete loss or overexpression; P0.001) of the primary colorectal cancer. In conclusion, the histological growth pattern in liver metastasis was a strong and independent prognostic factor for colorectal cancer. Our observations highlight the significant associations between histological growth patterns in liver metastases and histopathological features of the primary tumor, especially invasive front morphology and p53 aberration.

Ursodeoxycholic Acid Treatment Restores Gut Microbiota and Alleviates Liver Inflammation in Non-Alcoholic Steatohepatitic Mouse Model.

Gut microbiota dysbiosis plays an important role in the progression of non-alcoholic fatty liver disease (NAFLD), and no approved drugs are available for NAFLD treatment. In this study, we aimed to explore the dynamic changes of gut microbiota at the different stages of NAFLD and determine whether ursodeoxycholic acid (UDCA) could improve liver histopathological features of non-alcoholic steatohepatitis (NASH) mice induced by a high-fat high-cholesterol (HFHC) diet and its impact on gut microbiota. 6-week-old male C57BL/6 mice were fed with a HFHC or normal diet for 12, 18, and 24 weeks, respectively, to simulate the different stages of NAFLD. 16s ribosomal RNA genes from mice fecal samples at the different time points were sequenced to evaluate the dynamic changes of the gut microbiota. Then, C57BL/6 mice were fed with a HFHC diet for 24 weeks to establish the NASH model. Different doses of UDCA were administered intragastrically for additional 4 weeks. Normal diet–fed mice were taken as control. Serum samples, liver, and intestine tissues were harvested for biochemical tests and histopathological examinations. 16s ribosomal RNA genes from mice fecal samples were sequenced to assess the structural changes of gut microbiota. HFHC diet–fed mice developed simple steatosis, steatohepatitis, and fibrosis at 12, 18, and 24 weeks, respectively. The profile of gut microbiota dynamically changed with the different stages of NAFLD. NASH mice had significantly higher abundance of Fecalibaculum, Coriobacteriaceae_UCG-002, and Enterorhabdus, and lower abundance of norank_f_Muribaculaceae, Bacteroides, and Alistipes, which were partially restored by UDCA treatment. UDCA treatment significantly attenuated hepatic inflammation of NASH mice as indicated by the sum of ballooning and lobular inflammation of the NALFD activity score (3.2 ± 0.8 vs 1.8 ± 0.8, p = 0.029), and partially restored gut microbiota dysbiosis, and increased the expression of Claudin-1 and ZO-1 in the intestine, but did not activate the suppressed Farnesoid X receptor signal pathway. Conclusions: The gut microbiota dynamically changes with the different stages of NAFLD. UDCA treatment (120 mg/kg) could partially restore gut microbiota, repair gut barrier integrity, and attenuate hepatic inflammation in the NASH mouse model.

A novel minimally invasive OFM technique with orthotopic transplantation of hUC-MSCs and in vivo monitoring of liver metabolic microenvironment in liver fibrosis treatment

BackgroundMesenchymal stromal cells (MSCs) transplantation showed promising therapeutic results in liver fibrosis. However, efficient cell delivery method is urgently needed and the therapeutic mechanism remains unclear. This study focused on developing a minimally invasive open-flow microperfusion (OFM) technique, which combined orthotopic transplantation of human umbilical cord-derived (hUC)-MSCs to liver and in vivo monitoring of liver microenvironment in mice with CCl4-induced liver fibrosis.MethodsThe therapeutic potential of OFM route was evaluated by comparing OFM with intravenous (IV) injection route in terms of hUC-MSCs engraftment at the fibrosis liver, liver histopathological features, liver function and fibrotic markers expression after hUC-MSCs administration. OFM was also applied to sample liver interstitial fluid in vivo, and subsequent metabolomic analysis was performed to investigate metabolic changes in liver microenvironment.ResultsCompared with IV route, OFM route caused more hUC-MSCs accumulation in the liver and was more effective in improving the remodeling of liver structure and reducing collagen deposition in fibrotic liver. OFM transplantation of hUC-MSCs reduced blood ALT, AST, ALP and TBIL levels and increased ALB levels, to a greater extent than IV route. And OFM route appeared to have a more pronounced effect on ameliorating the CCl4-induced up-regulation of the fibrotic markers, such as α-SMA, collagen I and TGF-β. In vivo monitoring of liver microenvironment demonstrated the metabolic perturbations induced by pathological condition and treatment intervention. Two metabolites and eight metabolic pathways, which were most likely to be associated with the liver fibrosis progression, were regulated by hUC-MSCs administration.ConclusionThe results demonstrated that the novel OFM technique would be useful for hUC-MSCs transplantation in liver fibrosis treatment and for monitoring of the liver metabolic microenvironment to explore the underlying therapeutic mechanisms.

Supplementation of Choline-Deficient Diet With Pterostilbene Attenuates Cancer Development and Epigenetic Dysregulation of Gene Expression in Rat Livers

Abstract Objectives Nearly 40% of humans have polymorphisms in genes involved in choline metabolism which makes them prone to developing choline deficiency and increased risk for liver damage and liver cancer. Choline is a source of methyl groups needed for many steps in metabolism and epigenetic regulation of gene expression. Although epigenetic aberrations are known to be induced by choline deficiency, it remains unknown how to reverse the changes and attenuate symptoms. Interestingly, certain dietary compounds such as polyphenols have been demonstrated to reverse aberrant epigenetic patterns and exert anti-cancer action. In the present study, we investigate the effects of pterostilbene (PTS) on liver cancer development in rats fed choline-deficient diet and explore mechanisms underlying these effects. Methods Fischer 344 rats were fed a choline-sufficient (CSAA, healthy control group), a choline-deficient (CDAA, cancer group) L-amino acid-defined diet or a CDAA diet supplemented with PTS (134 mg/kg BW/day) (n = 6 per group). At the end of 52 weeks, analyses of liver nodules and histopathological features were performed followed by genome-wide investigation of gene expression in livers using RNA sequencing. DNA methylation was assessed by pyrosequencing. Results A total of 708 genes were significantly differentially expressed in CDAA + PTS group as compared with CDAA group. Among 351 upregulated genes were Bhmt (4.5-fold), G6pc (3.1-fold), and Aldh1l1 (2.6-fold). These metabolism-related genes were significantly downregulated in CDAA vs. CSAA group and their suppression was associated with liver cancer in previous reports. Among 357 genes found to be significantly downregulated by PTS were strong oncogenes such as Mmp12 (2-fold), Myc (1.9-fold) and Mmp27 (1.8-fold). We found PTS-mediated downregulation of Mmp12, that was a top gene upregulated in CDAA vs. CSAA, coincided with 43% hypermethylation of Mmp12 promoter. Conclusions Our findings demonstrate that PTS-mediated changes in gene expression could correspond to changes in DNA methylation of gene regulatory regions and could at least partially explain the observed attenuation of cancer development due to choline deficiency. Funding Sources UBC VP Academic Award, CFI John. R. Evans Leaders Fund, and BC Knowledge Development Fund granted to BS.