Pathological Changes Of Liver Tissues Research Articles

Effects of TUG1 on hepatic fibrosis and its mechanism

Objective: To investigate the effects and mechanisms of taurine up-regulated gene 1 (TUG1) in hepatic fibrosis. Methods: According to the literature, the classic hepatic fibrosis model of rats induced by 1%DMN(1ml/kg/d) was established. The rats with hepatic fibrosis and activated hepatic stellate cells (HSC) were divided into model control group, negative control group (transfected with siRNA negative control), siRNA interference group (transfected with TUG1). At the end of the experiment, hematoxylin eosin (HE) staining was used to detect the pathological changes of liver tissue; reverse transcription polymerase chain reaction (RT-PCR) and Western blot were used to determine the expression levels of α-smooth muscle actin (α-SMA), TUG1, collagen I, matrix metalloproteinase-2 (MMP-2), tissue inhibitor of metalloproteinase-1 (TIMP-1), Smad2 and Smad3 in rat liver tissue and activated hepatic stellate cells. Results: Compared with the model control group, the protein and gene levels of TUG1 and α-SMA in the negative control group were increased significantly(P＜0.05). The protein and gene levels of TUG1, α-SMA, collagen I, MMP-2, TIMP-1, Smad2 and Smad3 in the liver tissue and activated hepatic stellate cells in the siRNA interference group were decreased (P＜0.05) while compared with the blank control group and the negative control group. There were no significant differences in the levels of TUG1, α-SMA, collagen I, MMP-2, TIMP-1, Smad2 and Smad3 in the liver tissue and activated hepatic stellate cells between the control group and the negative control group (P＞0.05). Conclusion: TUG1 level is elevated in hepatic fibrosis tissue and activated hepatic stellate cells. Silencing TUG1 may improve the pathological damage of hepatic fibrosis induced by 1% DMN by inhibiting the transforming growth factor(TGF-β1)/ Smad signaling pathway.

The Intervention and Mechanism of Action for Aloin against Subchronic Aflatoxin B1 Induced Hepatic Injury in Rats.

As a class of difurancoumarin compounds with similar structures, aflatoxins (AF) are commonly found in the environment, soil, and food crops. AF pose a serious threat to the health of humans, poultry, and livestock. This study aimed to investigate the neuroprotective effect and detailed mechanism of aloin on hepatic injury induced by subchronic AFB1 in rats. The result showed that aloin could significantly inhibit the decrease in food intake, body weight growth, immune organ index, and serum albumin content caused by long-term AFB1 exposure. Meanwhile, aloin reduced the level of serum liver function and improved renal swelling and pathological changes of liver tissue. Aloin could also inhibit liver lipid peroxidation and improve liver antioxidant capacity. Further investigation revealed that aloin inhibited the activity and expression of hepatic CYP1A2 and CYP3A4 and down-regulated IL-1β expression in subchronic AFB1-induced liver injury rats. The above study demonstrated that aloin played an important role in blocking or delaying the development process of subchronic AFB1-induced hepatotoxicity. Therefore, aloin is considered to have a potential role as a protective agent against AFB1.

Lupeol reduces M1 macrophage polarization to attenuate immunologic dissonance and fatty acid deposition in rats with diet-induced metabolic syndrome.

BackgroundThis study aimed to investigate whether lupeol could inhibit the inflammatory mediators associated with the regulation of macrophage phenotypes and functions in rats with diet-induced metabolic syndrome (MS).MethodsForty specific-pathogen-free Sprague Dawley rats were fed a high-fat diet (HFD) for 10 weeks to establish an MS model. Lupeol was prepared and administered to the rats intraperitoneally at 20, 50, or 100 mg/kg (the lupeol 20 mg/kg, lupeol 50 mg/kg, and lupeol 100 mg/kg groups respectively). After 28 days of continuous intraperitoneal administration, rats were anesthesia and euthanasia. The obesity index, blood glucose and lipid metabolism indexes of rats in each group were measured. The levels of insulin and inflammatory factors in each group were detected by enzyme-linked immunosorbent assay (ELISA) kits. The pathological changes of liver tissue in rats were observed by hematoxylin and eosin (HE) and oil red O staining. The polarization levels of M1 and M2 macrophages in peripheral blood mononuclear cells (PBMCs) were analyzed by flow cytometry. The transcription levels of M1 and M2 macrophages markers were detected by qRT-PCR. The expressions of inducible nitric oxide synthase (iNOS) and arginase-1 (Arg-1) proteins in heart tissues of rats in each group were analyzed by Western blotting.ResultsLupeol significantly recovered fasting blood glucose and serum insulin levels, and reduced the production of proinflammatory cytokines, including interleukin (IL)-6, tumor necrosis factor alpha (TNF-α), and monocyte chemoattractant protein-1 (MCP-1), in the liver. It also elevated the expression of anti-inflammatory cytokines, including IL-4 and IL-10, in the MS model. Further, after treatment with lupeol, the levels of total cholesterol, triacylglycerol and low-density lipoprotein (LDL) were decreased, and high-density lipoprotein (HDL) were increased. Importantly, in the MS model group, lupeol remarkably inhibited M1 macrophages polarization (F4/80+iNOS+) while elevating M2 macrophages polarization (F4/80+CD206+) remarkably. At the same time, the levels of M1 markers, including inducible nitric oxide synthase, IL-1β, IL-6, and TNF-α, were markedly inhibited, while those of M2 markers, such as arginase-1, IL-10, CD206, and TGF-β, were markedly elevated in the MS model rats.ConclusionsLupeol might promote M2 polarization of macrophages to relieve damage caused by MS.

Effect of peroxisome proliferator-activated receptor-γ coactivator-1α on liver injury induced by periodontitis in rats.

To investigate the effect of peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) on liver injury induced by periodontitis in rats. Twenty-four male Wistar rats were randomly divided into two groups: control group and periodontitis group, twelve per group. In periodontitis group, the periodontitis models were established for the maxillary first molars in rats by means of "wire ligation+vaccinationwith Porphyromonas gingivalis", the control group was inoculated with the equal volume of 2% sodium carboxymethyl cellulose in the same position, for 6 weeks. The probing depth, tooth mobility and sulcus bleeding index were detected. Hematoxylin-eosin (HE) staining was used to observe the pathological changes of liver tissues in rats. The quantitative real-time polymerase chain reaction (qRT-PCR) and immunohistochemistry (IHC) were used to detect the gene and protein expression levels of PGC-1α, nuclear factor erythroid 2-related factor 2 (Nrf2) and mitochondrial transcription factor A (TFAM) in liver tissues of rats. The probing depth, tooth mobility and sulcus bleeding index in periodontitis group were significantly higher than that in control group. HE staining showed in periodontitis group, hepatic cords ranged disorderly and there were vacuoles in cells and inflammatory cells infiltrated in liver tissues of rats, and there was no obvious abnormality in control group. The qRT-PCR results showed that the mRNA expression levels of Pgc-1α, Nrf2 and Tfam in liver tissues of rats in periodontitis group were lower obviously than that in control group. IHC results showed that the protein expression level of PGC-1α in liver tissues of rats in periodontitis group was decreased significantly than that in control group. PGC-1α may be involved in the process of periodontitis-induced liver injury in rats.

Interplay Between Non-Canonical NF-κB Signaling and Hepatitis B Virus Infection.

The non-canonical nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) signaling pathway is an important component of NF-κB transcription complex. Activation of this pathway mediates the development and function of host immune system involved in inflammation and viral infection. During hepatitis B virus (HBV) infection, there is a complex interaction between infected hepatocytes and the immune cells, which can hinder antiviral immune responses and is associated with pathological changes in liver tissue. Consistently, the host immune system is closely related to the severity of liver damage and the level of viral replication. Previous studies indicated that the non-canonical NF-κB signaling pathway was affected by HBV and might play an important regulatory role in the antiviral immunity. Therefore, systematically elucidating the interplay between HBV and non-canonical NF-κB signaling will contribute the discovery of more potential therapeutic targets and novel drugs to treat HBV infection.

Baicalein pre-treatment alleviates hepatic ischemia/reperfusion injury in mice by regulating the Nrf2/ARE pathway.

Hepatic ischemia-reperfusion injury (HIRI) is caused by blood flow recovery following ischemia. Baicalein (BAI), a natural antioxidant used in traditional Chinese medicine, eliminates excessive free radicals and protects the structure of the cell membrane. However, its protective mechanism against HIRI is still unclear. The present study investigated underlying mechanism using a mouse HIRI model. Liver injury was evaluated using serum levels of alanine aminotransferase and aspartate aminotransferase, and hematoxylin-eosin staining was performed to evaluate the pathological changes in liver tissue. Apoptosis of hepatocytes was detected by TUNEL staining. The expression levels of reactive oxygen species (ROS), malondialdehyde (MDA) and superoxide dismutase (SOD) in the liver were detected to evaluate oxidative stress. Western blotting was performed to assess expression levels of nuclear factor E2-related factor 2 (Nrf2)/antioxidant response elements (ARE) pathway proteins in liver tissue. BAI pre-treatment significantly decreased elevation of serum aminotransferase levels induced by IR and alleviated histological damage to the liver. BAI decreased production of ROS and MDA in liver tissue induced by IR and increased the activity of SOD. At the same time, BAI inhibited apoptosis of liver cells induced by oxidative stress. Furthermore, BAI promoted the translocation of Nrf2 into the nucleus and increased the expression of total heme oxygenase-1 and NAD(P)H dehydrogenase quinone-1. The Nrf2 inhibitor ML385 reversed the protective effect of BAI on HIRI. These results indicated that BAI served a protective effect in HIRI by regulating the Nrf2/ARE pathway.

Aristolochic acid induces mitochondrial apoptosis through oxidative stress in rats, leading to liver damage

Aristolochic acid (AA) are persistent soil pollutants in the agricultural fields of the Balkan Peninsula. Preparations containing aristolochic acid are widely used for anti-inflammatory, diuretic, etc. To study the hepatotoxicity of aristolochic acid, 80 healthy SD rats were selected and divided into 20 mg/kg- AA group, 4 mg/kg-AA group, and 2 mg/kg-AA group and blank group, 20 rats per group. Mainly tested the body weight, liver function, liver tissue oxidative stress and pathological changes of liver tissue in rats. The ALT and AST activities in the serum of the rats in the administration groups were increased compared with the blank group. The activity of MDA in the administration groups was higher than that in the blank group; the activities of SOD, T-AOC and GSH-PX were significantly lower than those in the blank group. HE tissue sections also found that the administration groups showed varying degrees of hepatocyte boundary blur, nuclear fragmentation, and fibrosis tendency. Transmission electron microscopy showed that the mitochondria of the rat liver became more and more severely damaged with the increase of dose. Compared with the blank group, the mRNA expression of Bax, Caspase-9 and Caspase-3 in the administration groups were determined, while the mRNA expression of the Bcl-2 was increased. And compared with the blank control group, the expression levels of apoptotic proteins caspase-9 and caspase-3 increased significantly in the 20 mg/kg-AA group. Aristolochic acid can induce liver injury in rats through oxidative stress pathway and mitochondrial apoptosis pathway.

Analysis of liver pathological characteristics and exploration of noninvasive markers of liver fibrosis in children with chronic hepatitis B

Objective: To analyze and summarize the characteristics of liver pathology and their relation to clinical markers and further explore noninvasive markers of liver fibrosis in children with chronic hepatitis B. Methods: Data of 80 hospitalized children with chronic hepatitis B who underwent liver biopsy without antiviral treatment from 2011 to 2020 were retrospectively analyzed. Inflammation and liver fibrosis characteristics were analyzed in children of different ages and genders. Variables with good correlation with liver fibrosis stage were selected to establish a non-invasive diagnostic score of liver fibrosis in children. Measurement data was used to compare the t-test or rank sum test. Mantel-Haenszel χ (2) test was used for bidirectional ordered grouping data. Spearman's rank correlation test was used for rank correlation analysis. Receiver operating characteristic curve (ROC) was used to evaluate the diagnostic value of the newly established diagnostic score in children with liver fibrosis. Results: The median age of the children was 6.4 years. HBV DNA level was high (P50 = 7.6 log(10) IU/ml), and serum alanine aminotransferase (ALT) in P50 was 171 U/L (< ULN: 5 cases, ULN-2ULN: 10 cases, > 2 ULN: 65 cases). Pathological analysis showed that the incidence of liver tissue inflammation was 97.5%, and the proportion of patients with G≥2 was 42.5%, while S≥2 was 36.3%. The incidence rate of liver fibrosis and liver cirrhosis was 81.3%, and 1.3%, respectively. The changes in liver tissue inflammation and fibrosis were gradually aggravated with the increase of age, and the proportion of high-grade inflammation and liver fibrosis in male children was higher than that in female children. Serum levels of glutamyl transpeptidase (GGT), γ-glutamyltransferase/platelet ratio (GPR) and HBeAg had a good correlation with fibrosis stage (r(s) = 0.397, 0.389, and - 0.311) in children with chronic hepatitis B. The combination of GGT, GPR and HBeAg can establish a non-invasive diagnostic score for evaluating liver fibrosis in children. When the score is less than 1.5, it can be diagnosed as S0, and 1.5 ≤ score < 3.5, it can be diagnosed as S1; 3.5 ≤ score < 5.5, the diagnosis of fibrosis is S2; score≥ 5.5, the diagnosis of fibrosis is S≥3. The sensitivity and specificity were 80%, 83%, 86%, and 53%, 55%, 67%, respectively. Conclusion: The incidence of liver tissue inflammation in children with chronic hepatitis B with elevated and fluctuating transaminase levels is high, and the pathological changes of liver tissue aggravate with the age of the children. GGT, GPR and HBeAg have a good correlation with liver fibrosis in children with chronic hepatitis B. Therefore, combining the above-mentioned markers to establish a new noninvasive diagnostic score has certain diagnostic value for liver fibrosis stage S0-S3 in children with chronic hepatitis B.

Effects of evodiamine on carbon tetrachloride-induced liver fibrosis mice based on modulating gut microbiota

Objective: To access the effects of evodiamine on carbon tetrachloride (CCl(4)) -induced liver fibrosis mice and study the mechanism based on modulating gut microbiota. Methods: From August 2019, 30 SPF male C57BL/6 mice were randomly divided into normal, model and evodiamine groups. Mice in control group received intraperitoneal injection of olive oil (2 ml/kg, twice per week) for 6 weeks. Mice in model and evodiamine groups received intraperitoneal injection of 20% CCl(4) (2 ml/kg, twice per week) for 6 weeks to induce liver fibrosis mice. Then, mice in evodiamine group received orally of evodiamine (18 mg/kg) for 4 weeks. The levels of serum alanine aminotransferase (ALT) , aspartate aminotransferase (AST) , albumin (ALB) and total protein (TP) were detected. The pathological changes of liver tissue were observed. The effects of evodiamine on the abundance and diversity of intestinal microflora in liver fibrosis mice were determined. The mRNA and protein expression levels of inflammatory factors[interleukin-6 (IL-6) , interleukin-1β (IL-1β) , and tumor necrosis factor α (TNF-α) ] in liver tissue were measured. Results: Compared with the normal group, the body weight, serum ALB and TP levels of the model group were decreased, the liver index, ALT and AST levels were increased, and the intestinal flora Shannon and Simpson indexes were decreased (P<0.01) . Compared with the normal group, the abundance of Lactobacillus, Akkermansia and Bacteroides in the feces of the model group decreased, while the abundance of Enterococcus and Lachnoclostridium increased (P<0.01) . Compared with the normal group, the mRNA and protein expressions levels of IL-6, IL-1β, and TNF-α in the liver tissue of the model group were significantly increased (P<0.01) . Compared with the model group, evodiamine could reduce liver index and serum ALT and AST levels, increase ALB and TP levels (P<0.05) , improve inflammatory cell infiltration and fibrosis degree in liver tissue, and up regulate intestinal flora Shannon and Simpson indexes in liver fibrosis mice (P<0.05) . Compared with the model group, evodiamine could increase the abundance of Lactobacillus, Akkermansia, Bacteroides, and reduce the abundance of Enterococcus and Lachnoclostridiun (P<0.05) . Compared with the model group, evodiamine could reduce the mRNA and protein levels of IL-6, IL-1β and TNF-α in liver tissue of liver fibrosis mice (P<0.05) . Conclusion: Evodiamine can ameliorate CCl(4)-induced liver fibrosis through modulating gut microbiota and inhibiting the inflammatory response in liver.

Schisantherin A ameliorates liver fibrosis through TGF-β1mediated activation of TAK1/MAPK and NF-κB pathways in vitro and in vivo

BackgroudSchisandra chinensis, a traditional Chinese medicine for liver protection, can significantly improve liver fibrosis. However, it is still unclear which active components in Schisandra chinensis play an anti-fibrosis role. PurposeThe purpose of present study was to observe the anti-fibrosis effect of schisantherin A (SCA) on liver fibrosis and explore its underlying mechanism. MethodsThe liver fibrosis model of mice was constructed by the progressive intraperitoneal injection of thioacetamide (TAA), and SCA (1, 2, and 4 mg/kg) was administered by gavage for 5 weeks. The biochemical indicators and inflammatory cytokines were measured, changes in the pathology of the mice liver were observed by hematoxylin & eosin (H&E) and Masson stainings for studying the anti-fibrosis effect of SCA. A hepatic stellate cell (HSCs) activation model induced by transforming growth factor-β1 (TGF-β1) was established, and the effect of SCA on the HSCs proliferation was observed by MTT assay. The expressions of target proteins related to transforming growth factor-β-activated kinase 1 (TAK1)/mitogen-activated protein kinase (MAPK) and nuclear factor-κB (NF-κB) pathways were evaluated by western blotting, immunohistochemistry or immunofluorescence analysis, to explore the potential mechanism of SCA. ResultsSCA could significantly ameliorate the pathological changes of liver tissue induced by TAA, and reduce the serum transaminase level, the hydroxyproline level and the expression of α-smooth muscle actin (α-SMA) and collagen 1A1 (COL1A1) proteins in the liver tissue. SCA could significantly lower the levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6) in the serum and liver tissue, and down-regulate the expression of target proteins related to TAK1/MAPK and NF-κB pathways in the liver tissue. The in vitro studies demonstrated that SCA significantly inhibited the proliferation and activation of HCS-T6 cells induced by TGF-β1, decreased TNF-α and IL-6 levels, and inhibited the TAK1 activation induced by TGF-β1 and then the expression of MAPK and NF-κB signaling pathway-related proteins. ConclusionTogether, SCA can ameliorate the liver fibrosis induced by TAA and the HSC-T6 cell activation induced by TGF-β1 in mice, and its mechanism may be to inhibit the HSCs activation and inflammatory response by inhibiting TGF-β1 mediated TAK1/MAPK and signal pathways.

Monofloral Triadica Cochinchinensis Honey Polyphenols Improve Alcohol-Induced Liver Disease by Regulating the Gut Microbiota of Mice.

Honey produced from medicinal plants holds great promise for human health. Increasing evidence suggests that the gut microbiota plays an important role in liver pathology after alcohol intake. The aim of this study was to identify the polyphenol composition of triadica cochinchinensis honey (TCH), and to study the potential effect of honey polyphenols on the regulation of gut microbes in mice with alcohol-induced liver injury and the improvement of alcohol-induced liver disease. For these purposes, a total of 190 compounds were identified and 27 of them were quantified by ultraperformance liquid chromatography coupled with quadrupole/time-of-flight mass spectrometry (UPLC-Q/TOF-MS) and we successfully established a mouse model of alcohol-induced liver injury. The results show that TCH polyphenols can significantly restore the levels of ALT and AST, and TCH intervention can significantly improve the pathological changes of liver tissue in alcohol-exposed mice. Additionally, a significant decrease was observed in Firmicutes/Bacteroidetes after TCH treatment. Moreover, KEGG pathways of ATP-binding cassette (ABC) transporters, two-component system and biosynthesis of amino acids enriched the most differentially expressed genes after TCH intervention for 8 weeks. Our results may have important implications for the use of TCH as a functional food component with potential therapeutic utility against alcohol-induced liver disease.

Platycodon grandiflorus Polysaccharide with Anti-Apoptosis, Anti-Oxidant and Anti-Inflammatory Activity Against LPS/D-GalN Induced Acute Liver Injury in Mice

This study aimed to explore the mechanism of hepatoprotective effect of Platycodon grandiflorus polysaccharides (PGPSt) on LPS/D-galactose (D-GalN)-induced acute liver injury in mice. The pathological changes of liver tissue were observed by H&E stain. The results showed that the structure of hepatocytes in the model group was destroyed obviously, and that in 200 mg/kg PGPSt group was improved, which indicated that PGPSt could reduce the damage of hepatocytes. Then, indicators of oxidative stress, inflammatory cytokines and apoptosis were detected. The results showed that PGPSt significantly reduced the activities of AST, ALT and SOD. PGPSt increased the expression of GSH, and decreased the level of MDA, IL-6, IL-1β and TNF-α. PGPSt can inhibit hepatocyte apoptosis, which might be related to down-regulation of cleaved-caspase 3 and Bax, and up-regulation of Bcl-2. In addition, Western blot analyses revealed that PGPSt suppressed the protein expression of TRL4, p-P38 and NF-κB p65. PGPSt exhibited protective effects on LPS/D-GalN-induced acute liver injury in mice. Conclusion, the results demonstrated that PGPSt is a potential therapeutic drug by alleviating oxidative stress, reducing inflammatory cytokines and enhancing the expression of anti-apoptotic proteins that could be used for the treatment of some acute liver injury in the future.

Effect of total saponins from Panax japonicus on non-alcoholic steatohepatitis by regulating autophagy

Non-alcoholic steatohepatitis(NASH) was induced by high-sugar and high-fat diet in mice to investigate the intervention effect of total saponins from Panax japonicus(TSPJ) and explore its possible mechanism. Mice were fed with high-sugar and high-fat diet to establish NASH model, and intervened with different doses of TSPJ(15, 45 mg·kg~(-1)). The animals were fed for 26 weeks. The histomorphology and pathological changes of liver tissues were observed by HE staining. The transcriptional expression levels of miR-199 a-5 p, autophagy related gene 5(ATG5) and inflammatory cytokines interleukin-6(IL-6), interleukin-1β(IL-1β) and tumor necrosis factor α(TNF-α) in mouse liver were measured by quantitative Real-time polymerase chain reaction(qRT-PCR). Western blot was used to detect the expression of autophagy-related proteins ATG5, P62/SQSTM1(P62), and microtubule-associated protein light chain 3(LC3)-I/Ⅱ proteins in mouse liver. The expression of P62 protein was detected by immunofluorescence staining. In order to verify the targeting regulation relationship between miR-199 a-5 p and ATG5, miR mimic/inhibitor NC and miR-199 a-5 p mimic/inhibitor were transfected into Hepa 1-6 cells, and the expression of ATG5 mRNA and protein was detected. pMIR-reportor ATG5-3'UTR luciferase reporter gene plasmid was constructed and co-transfected with miR mimic/inhibitor NC and miR-199 a-5 p mimic/inhibitor into Hepa 1-6 cells to detect luciferase activity. In vivo, HE staining in the model group showed typical fatty degeneration and inflammatory infiltration, with increased expression of miR-199 a-5 p and decreased expression of ATG5 mRNA and protein. The expression of autophagy-associated protein P62 increased significantly, the ratio of LC3Ⅱ/Ⅰ decreased, and the transcriptional expression of inflammatory factors increased significantly. After the intervention by TSPJ, the pathological performance of liver tissue was significantly improved, the expression of miR-199 a-5 p decreased and the expression of ATG5 mRNA and protein increased, the expression of autophagy-associated protein P62 decreased significantly, the ratio of LC3Ⅱ/Ⅰ increased, and the transcriptional expression of inflammatory cytokines IL-6, IL-1β and TNF-α decreased significantly. In vitro, it was found that the expression of ATG5 mRNA and protein and luciferase activity decreased significantly in miR-199 a-5 p overexpression cells, while after inhibition of miR-199 a-5 p expression, the expression level of ATG5 mRNA and protein and luciferase activity increased. The results showed that TSPJ can improve NASH in mice fed with high-sugar and high-fat diet, and its mechanism may be related to the regulation of miR-199 a-5 p/ATG5 signal pathway, the regulation of autophagy activity and the improvement of inflammatory response of NASH.

Light optical and Ultrastructural Characteristics of the Maral Liver with Chronic Dicrocoeliasis

BACKGROUND: Dicrocoeliasis is caused by trematode Dicrocoelium lanceatum from the family Dicrocoeliidae, a parasite in the bile ducts of the liver of domestic and wild animals. Dicrocoeliasis mainly affects sheep, cattle, camels, zebu, deer, fallow deer, argali, less often – horses, donkeys, dogs, rabbits, hares and bears, as well as humans. Dicrocoeliasis of ruminants is widespread across the whole Kazakhstan. Invasive diseases represent a significant obstacle in the development of domestic maral breeding, among which trematodoses, and particularly dicrocoeliasis of maral play a major role. AIM: The aim of the research was to study the influence of dicrocoelia on the ultrastructural organization of the liver of maral. MATERIALS AND METHODS: For examination under electron microscope, biopsy pieces of liver tissue of maral were fixed in 2.5% solution of glutaraldehyde with post-fixation in 1% solution of osmium tetroxide, conducted according to a conventional method, and enclosed in epon. Semi-thin and ultra-thin sections were prepared on the ultra-microtome Leica. Semi-thin sections were stained with methylene blue, azure 2, and studied at a high-resolution light optical level. The ultra-thin sections were contrasted with uranyl acetate and lead citrate according Reynolds method and examined under electron microscope Libra 120 (C. Zeiss). RESULTS: The light optical examination of half-thin sections revealed that the morphological pattern of pathological changes in liver tissue was polymorphic, even within a single hepatic lobe. CONCLUSIONS: In the liver of maral infected with chronic dicrocoeliasis, dystrophic and destructive pathological changes developed in all the cellular structures of the hepatic lobules: In the form of plethora and vast enlargement of sinusoids, vacuolar and lipodegeneration of hepatocytes, destruction of the hepatic tissue with edema, hemorrhages, in the appearance of cells associated with inflammation, and the deposition of hematin crystals.

Effects of Danhong injection on dyslipidemia and cholesterol metabolism in high-fat diets fed rats

Ethnopharmacological relevanceDanhong injection (DHI) is a Chinese medical injection applied to the clinical treatment of cardiovascular diseases that has anti-inflammatory, antiplatelet aggregation and antithrombotic effects. This study aimed to explore the effects of DHI on dyslipidemia and cholesterol metabolism in high-fat diet-fed rats. MethodsSprague Dawley (SD) rats were randomly divided into six groups: normal group (Normal); hyperlipidemia model group (Model); DHI-treated groups at doses of 1.0 mL/kg, 2.0 mL/kg, 4.0 mL/kg; and simvastatin positive control group (2.0 mg/kg). The hypolipidemic effects of DHI were evaluated by measuring serum lipid levels, hepatic function and oxidative stress, respectively. And pathological changes in liver tissues were determined using hematoxylin-eosin (H&E) and oil red O staining. Moreover, the mRNA and protein expression levels of cholesterol metabolism related genes were detected by real-time PCR (RT-PCR) and Western blot. ResultsCompared with the Model group, DHI treatment markedly decreased the liver index and improved the pathological morphology of liver tissues. DHI treatment dose-dependently decreased the levels of total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), malondialdehyde (MDA), and free fatty acids (FFA) in serum or liver tissues (P < 0.01 or P < 0.05), and increased the high-density lipoprotein cholesterol (HDL-C) and tripeptide glutathione (GSH) (P < 0.01 or P < 0.05). The activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX) were increased in the DHI-treated groups (P < 0.01 or P < 0.05), while the alanine transaminase (ALT) and aspartate transaminase (AST) were decreased (P < 0.01 or P < 0.05). Furthermore, the expression levels of LDL receptor (LDLR), cholesterol 7-α-hydroxylase (CYP7A1), liver X receptor α (LXRα), and peroxisome proliferator-activated receptor α (PPARα) were dose-dependently upregulated in the DHI-treated groups, whereas the expression of sterol regulatory element-binding protein-2 (SREBP-2) was downregulated. ConclusionsOur study demonstrated that DHI markedly ameliorated hyperlipidemia rats by regulating serum lipid levels, inhibiting hepatic lipid accumulation and steatosis, improving hepatic dysfunction, and reducing oxidative stress. The potential mechanism was also tentatively investigated and may be related to the promotion of bile acid synthesis via activation of the PPARα-LXRα-CYP7A1 pathway. Therefore, DHI could be regarded as a potential hypolipidemic drug for the treatment of hyperlipidemia.

SCARF1 promotes M2 polarization of Kupffer cells via calcium-dependent PI3K-AKT-STAT3 signalling to improve liver transplantation.

ObjectivesThis study aimed to investigate the protective effect of SCARF1 on acute rejection (AR), phagocytic clearance of Kupffer cells (KCs), M2 polarization and the exact mechanism underlying these processes.MethodsAAV was transfected into the portal vein of rats, and AR and immune tolerance (IT) models of liver transplantation were established. Liver tissue and blood samples were collected. The level of SCARF1 was detected via WB and immunohistochemical staining. Pathological changes in liver tissue were detected using HE staining. Apoptotic cells were detected using TUNEL staining. KC polarization was assessed via immunohistochemical staining. Primary KCs were isolated and co‐cultured with apoptotic T lymphocytes. Phagocytosis of apoptotic cells and polarization of KCs were both detected using immunofluorescence. Calcium concentration was determined using immunofluorescence and a fluorescence microplate reader. The levels of PI3K, p‐AKT and P‐STAT3 were assessed via WB and immunofluorescence.ResultsCompared to the IT group, the level of SCARF1 was significantly decreased in the AR group. Overexpression of SCARF1 in KCs improved AR and liver function markers. Enhanced phagocytosis mediated by SCARF1 is beneficial for improving the apoptotic clearance of AR and promoting M2 polarization of KCs. SCARF1‐mediated enhancement of phagocytosis promotes increased calcium concentration in KCs, thus further activating the PI3K‐AKT‐STAT3 signalling pathway.ConclusionsSCARF1 promotes the M2 polarization of KCs by promoting phagocytosis through the calcium‐dependent PI3K‐AKT‐STAT3 signalling pathway.

Alprostadil alleviates liver injury in septic rats via TLR4/NF-κB pathway.

The aim of this study was to explore the role of alprostadil (Alp) in cecal ligation and puncture (CLP)-induced septic injury in rats and its possible mechanism of action. Wistar rats were randomly assigned into three groups, including: Sham group (no CLP was performed), CLP group (CLP was conducted) and Alp group (Alp was injected after CLP). Serum liver function markers, pathological changes in liver tissues, alterations in the level of oxidative stress, activity of the Toll-like receptor 4 (TLR4)/nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) pathway, and release of inflammatory factor tumor necrosis factor alpha (TNF-α) in the liver tissue homogenate were detected in each group. Compared with Sham group, the rats in CLP group had substantially elevated content of serum liver function markers, increased apoptotic liver cells, upregulated levels of oxidative stress, enhanced activity of the TLR4/NF-κB pathway, and increased release of TNF-α (p<0.05). Meanwhile, there were evident pathological changes under microscopic examination in CLP group compared with Sham group (p<0.05). In comparison with CLP group, Alp group exhibited significantly decreased concentrations of liver function markers, microscopic findings, such as decreased inflammatory cell infiltration in the interstitum, notably lowered proportion of apoptotic cells, decreased level of oxidative stress, weakened activity of the TLR4/NF-κB pathway and restrained release of TNF-α (p<0.05). Furthermore, normal morphology of liver cells was observed in Alp group compared with CLP group (p<0.05). Alp alleviates liver injury in septic rats by inhibiting the TLR4/NF-κB pathway.

C-Myc affects hedgehog pathway via KCNQ1OT1/RAC1: A new mechanism for regulating HSC proliferation and epithelial-mesenchymal transition

BackgroundThis study aimed to probe into the potential mechanism of KCNQ1OT1 in liver fibrosis. MethodsThe pathological changes in liver tissues were observed by Masson and hematoxylin-eosin (HE) staining. The proliferation or cell cycle of hepatic stellate cells (HSCs) was analyzed by MTT or flow cytometry. The expressions of epithelial markers E-cadherin, interstitial markers Snail and Vimentin, and hedgehog signaling pathway-related molecules Hhip, Shh, and Gli2 were detected by Western blot. The interaction or binding of c-Myc with the KCNQ1OT1 promoter was analyzed by dual-luciferase reporter gene or Chromatin immunoprecipitation (ChIP)-qPCR, and the interaction between KCNQ1OT1 and RAC1 was assessed by RNA immunoprecipitation and RNA pull-down. Moreover, the stability of RAC1 protein was detected by cycloheximide-chase and ubiquitination. Resultsc-Myc and KCNQ1OT1 were up-regulated in liver fibrosis tissues and cells. After the interference with c-Myc in primary-1-Day HSCs, the down-regulated KCNQ1OT1 restrained HSC proliferation and EMT by down-regulating RAC1 expression and restraining the hedgehog pathway. ConclusionOur results indicated that the interference with c-Myc down-regulated RAC1 expression and restrained the hedgehog pathway by down-regulating KCNQ1OT1, thus restraining HSC proliferation and EMT in liver fibrosis.

A Study on the Therapeutic Efficacy of San Zi Yang Qin Decoction for Non-Alcoholic Fatty Liver Disease and the Underlying Mechanism Based on Network Pharmacology.

Objective This study aims to explore the therapeutic efficacy of San Zi Yang Qin Decoction (SZ) and its potential mechanism in the treatment of non-alcoholic fatty liver disease (NAFLD) based on network pharmacology and in vivo experiments. Methods Effective chemicals and targets of SZ were searched in online databases, according to the drug-likeness of compounds and the binomial distribution of targets. A disease-target-chemical network was established using NAFLD-associated genes screened through GeneCards database, Gene Ontology (GO) terms, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. Furthermore, animal experiments were conducted to verify the efficacy and mechanism of SZ predicted by network pharmacology. The NAFLD mouse model was established with C57BL/6J mice fed with a high-fat diet for 22 weeks. The mice in the control group were fed with a chow diet. From the 23rd week, the NAFLD mice were treated with intragastric SZ or normal saline for 8 weeks. After the glucose tolerance was measured, the mice were sacrificed, followed by the collection of serum and liver tissues. Pathological changes in liver tissues were examined by H&E staining. Additionally, alanine aminotransferase (ALT), aspartate aminotransferase (AST), serum fast blood glucose, and insulin levels were detected. Expression levels of TNF-α of serum and liver tissues were determined by ELISA and qRT-PCR, respectively. Western blot was used to detect the activation of AKT in liver tissues. Results A total of 27 effective compounds and 20 targets of SZ were screened. GO analysis uncovered a significant correlation between the targets of SZ and those of NAFLD. KEGG analysis presented the signaling pathways enriched in SZ and NAFLD, including NAFLD, TNF-α, and apoptosis pathways. The area under the curve of major GO and KEGG pathways indicated the potential role of SZ in improving NAFLD. In vivo experiments demonstrated that SZ significantly alleviated hepatosteatosis and inflammatory cell infiltration in liver tissues, reduced serum transaminases, and improved insulin resistance and glucose tolerance of NAFLD mice. The protein level of phospho-AKT was upregulated by SZ. Additionally, SZ treatment obviously impaired the TNF-α level in the serum and liver tissue of NAFLD mice. Conclusions According to the network pharmacology analysis and in vivo experiments, SZ could have therapeutic efficacy for NALFD. The mechanism mainly involves pathways relative to insulin resistance, TNF-α, and apoptosis. Our results provide a scientific basis for SZ in the clinical treatment of NAFLD.

TLR4-MyD88-NF-κB signaling pathway contributes to the progression of secondary hepatic injury and fibrosis in hepatolithiasis

This paper focused on evaluating the effect of TLR4-MyD88-NF-κB signaling pathway in the progression of secondary hepatic injury and fibrosis in hepatolithiasis. The levels of inflammatory factors (IL-1β, IL-6, TNF-α) and serum biochemical values (ALT, AST, Tbil, Dbil, ALP, GGT) were detected by ELISA. IHC was used to detected the expression level of TLR4 in liver tissues of hepatolithiasis patients and mice. The pathological changes of liver tissue were observed by HE staining. The levels of MyD88, NF-κB, IκB, Laminin (LN), and chitosan enzyme 3-like protein 1 (CHI3L1) were detected by western blotting. In hepatolithiasis patients, the levels of TNF-α, IL-1β, and IL-6 were distinctly raised and proteins associated with TLR4-MyD88-NF-κB signaling pathway (such as TLR4, MyD88, NF-κB, and IκB) in liver tissues were significantly up-regulated. In Bile duct ligation (BDL) model of mice, the results showed that in addition to the significant increase of inflammatory factors, liver function indexes, and fibrosis indexes in BDL mice were also significantly up-regulated. Additionally, TLR4-MyD88-NF-κB signaling pathway was activated in BDL mice. After TLR4 knockdown in BDL mice, inflammatory factors, liver function indexes, and fibrosis indexes were significantly down-regulated. TLR4-MyD88-NF-κB signaling pathway proteins were restrained. TLR4-MyD88-NF-κB signaling pathway took part in the progression of secondary hepatic injury and fibrosis in hepatolithiasis. Inhibition of TLR4-MyD88-NF-κB signaling pathway can reduce the progression of secondary hepatic injury and fibrosis in hepatolithiasis.