Hepatic Fibrosis Tissues Research Articles

Mitochondria-Targetable Near-Infrared Fluorescent Probe for Visualization of Hydrogen Peroxide in Lung Injury, Liver Injury, and Tumor Models.

Hydrogen peroxide (H2O2) overexpressed in mitochondria has been regarded as a key biomarker in the pathological processes of various diseases. However, there is currently a lack of suitable mitochondria-targetable near-infrared (NIR) probes for the visualization of H2O2 in multiple diseases, such as PM2.5 exposure-induced lung injury, hepatic ischemia-reperfusion injury (HIRI), nonalcoholic fatty liver (NAFL), hepatic fibrosis (HF), and malignant tumor tissues containing clinical cancer patient samples. Herein, we conceived a novel NIR fluorescent probe (HCy-H2O2) by introducing pentafluorobenzenesulfonyl as a H2O2 sensing unit into the NIR hemicyanine platform. HCy-H2O2 exhibits good sensitivity and selectivity toward H2O2, accompanied by a remarkable "turn-on" fluorescence signal at 720 nm. Meanwhile, HCy-H2O2 has stable mitochondria-targetable ability and permits monitoring of the up-generated H2O2 level during mitophagy. Furthermore, using HCy-H2O2, we have successfully observed an overproduced mitochondrial H2O2 in ambient PM2.5 exposure-induced lung injury, HIRI, NAFL, and HF models through NIR fluorescence imaging. Significantly, the visualization of H2O2 has been achieved in both tumor-bear mice as well as surgical specimens of cancer patients, making HCy-H2O2 a promising tool for cancer diagnosis and imaging-guided surgery.

Effect of curcumin on lipid profile, fibrosis, and apoptosis in liver tissue in abemaciclib-administered rats

Abemaciclib (ABEM) is an important antitumor agent for breast cancer treatment. However, the side-effects of ABEM are unclear in the liver. This study investigated the protective effect of curcumin (CURC) on liver damage caused by ABEM. The rats were divided into five groups with eight animals in each group; Control, DMSO (150 µL for per rats), CURC, 30 mg/kg/day), ABE (26 mg/kg/day), and ABE + CURC (26 mg/kg/day ABE, 30 mg/kg/day) groups. Injections were administered daily for 28 days. The levels of AST, LDH, HDL, LDL, triglyceride, and total cholesterol in serum, and hepatic tissue fibrosis, caspase-3, Bax, and TNF-α expression were higher in the ABE group compared to the control group (p < 0.05). Also, these parameters in the ABEM + CURC group were lower than in the ABE group (p < 0.05). The results showed that ABE administration could cause liver damage and increase fibrosis in the liver. In addition, it was shown that co-administration of CURC with ABE could suppress the levels of AST, LDH, HDL, LDL, triglyceride, and total cholesterol in serum, and fibrosis, caspase-3, Bax, and TNF-α expressions in the liver. These data are the first in the literature. Therefore, the administration of CURC following ABE may be a therapeutic agent in preventing liver damage.

The Combination of Mesenchymal Stem Cells and Bovine Colostrum in Reducing α-SMA Expression and NLR Levels in Wistar Rats After 50% Fibrotic Liver Resection

Background: Liver fibrogenesis will produce α-smooth muscle actin (α-SMA) expression and a continuous inflammatory process, seen through the neutrophil lymphocyte ratio (NLR). The combination of mesenchymal stem cells and bovine colostrum is a novel strategy for repairing hepatic fibrosis tissue. To assess the combination of mesenchymal stem cells and bovine colostrum to reduce α-SMA expression and NLR levels in Wistar rats after 50% fibrotic liver resection.&#x0D; Methods: Thirty-six Wistar male rats were randomly divided into 6 groups (sham, control, colostrum, MSCs, and colostrum and MSCs combination). Rats were injected with CCl4 for 8 weeks to induce liver fibrosis then underwent liver resection. NLR levels was determined using Hematology Analyzer, α-SMA expression of myofibroblast was analyzed by immunofluorescence staining.&#x0D; Results: A significant reduction in NLR levels on day 3 in the treatment group I (1.10), treatment II (0.83), treatment III (0.93) compared to the control group. A significant reduction in NLR levels on day 10 in the treatment group I (0.76), treatment II (0.64), treatment III (0.54) compared to the control group. A significant decrease in α-SMA in treatment group I (0.134), treatment II (0.68), treatment III (0.42) compared to the control group.&#x0D; Conclusion: In this study, it was found that α-SMA expression, NLR levels on the 3rd and 10th day of administration were reduced in group receiving combination of mesenchymal stem cells and bovine colostrum in the liver of post-resection Wistar rats by 50%.

The anti-inflammatory effect of Pien Tze Huang in non-alcoholic fatty liver disease

Nonalcoholic fatty liver disease (NAFLD) is the most common liver disease that may progress to nonalcoholic steatohepatitis (NASH), hepatic tissue fibrosis, liver cirrhosis, and hepatocellular carcinoma. In this study, we investigated the effects of Pien Tze Huang (PTH), a well-known traditional Chinese herbal formula with liver protective effect, in methionine-choline deficient diet (MCD)- and high-fat diet (HFD)-induced NASH mouse models. Our results showed that PTH could exert hepatoprotective effects by improving liver weight and steatosis and reducing the fibrosis and serum levels of alanine transaminase (ALT) and aspartate transaminase (AST) in both animal models. The effects of PTH was accompanied with the reduction of infiltrated macrophages, the inhibition of the expression of cytokines, and the induction of adiponectin expression. Mechanistically, we found that PTH could inhibit the activation of proinflammatory transcription factor nuclear factor-κB (NF-κB) by preventing the degradation of inhibitor of κBα (IκBα). These results demonstrate that PTH can improve NAFLD largely due to its suppression of the NF-κB inflammatory pathway.

EVALUATION OF THE HEPATOTOXIC EFFECT OF LONG-TERM USE OF STATINS BASED ON THE RESULTS OF SHEAR WAVE ELASTOMETRY

Statins have been used in clinical practice for more than a quarter of a century. Over this period, their positions in various clinical guidelines have significantly strengthened. First, this group of drugs was used exclusively for the treatment of patients with a pronounced increase in the levels of atherogenic lipoproteins, but over time, the indications for their prescription have expanded considerably as well as the tactics of their use. The aim of our work is to determine the effect of statin therapy on hepatic tissue fibrosis in patients with steatohepatosis. We examined 87 patients (43 women and 44 men) with non-alcoholic steatohepatosis and liver fibrosis. The average age of the patient was 46.2 ± 4.6 years. The studies were carried out at the scientific and practical centre of the Department of Family Medicine and Therapy, Poltava State Medical University, and the Rheumatology Centre, Poltava Region Clinical Hospital. The severity of liver fibrosis was established based on 2D shear wave elastometry by transcutaneous shear wave approach in SWE mode using a convex transducer tuned to 3.5 MHz on an Ultima PA Expert ultrasound scanner (Radmir, Ukraine). The level of "stiffness" of the liver parenchyma by SWE was measured in kPa. The study included patients with liver fibrosis F2-F3 according to METAVIR. All patients received statin therapy during the study: 22 patients received rosuvastatin at a daily dose of 20 mg, 21 patients received atorvastatin at a daily dose of 20 mg, 22 patients received simvastatin at a daily dose of 20 mg, and 22 patients received lovastatin in a daily dose of 20 mg. In addition to a decrease in the level of low density lipoproteins, total cholesterol, triglycerides, systemic and long-term use of statins under the adequate treatment of steatohepatosis has led to a decrease in the severity of fibrotization of the hepatic parenchyma that is confirmed by both shear wave elastometry data and the scale data. At the same time, the difference between the indicators after the therapy in the groups of patients receiving rosuvastatin, simvastatin, atorvastatin and lovastatin showed no signs of reliability that may indicate a positive effect of inhibition of hydroxymethylglutaryl coenzyme A reductase on the progression of fibrotic changes. The use of statins in the integrated therapy of patients with steatohepatitis, in addition to normalizing the plasma lipid profile, can reduce the fibrotic changes in the liver parenchyma. It should be noted that there is no advantage to any of the medicines we investigated. Moreover, the statins used did not produce hepatotoxic effects.

Ruxolitinib suppresses liver fibrosis progression and accelerates fibrosis reversal via selectively targeting Janus kinase 1/2

BackgroundJAK1 and JAK2 have been implicated in fibrosis and cancer as a fibroblast-related marker; however, their role in liver fibrosis has not been elucidated. Here, we aim to determine the effect and underlying mechanism of JAK1/2 inhibition on liver fibrosis and hepatic stellate cells (HSCs) and further explore the therapeutic efficacy of Ruxolitinib, a JAK1/2 selective inhibitor, on preventing and reversing liver fibrosis in mice.MethodsImmunohistochemistry staining of JAK1 and JAK2 were performed on liver tissue in mice with hepatic fibrosis and human liver tissue microarray of liver cirrhosis and liver cancer. LX-2 cells treated with specific siRNA of JAK1 and JAK2 were used to analysis activation, proliferation and migration of HSCs regulated by JAK1/2. The effects of Ruxolitinib (JAK1/2 inhibitor) on liver fibrosis were studied in LX-2 cells and two progressive and reversible fibrosis animal models (carbon tetrachloride (CCl4), Thioacetamide (TAA)).ResultsWe found that JAK1/2 expression was positively correlated with the progression of HCC in humans and the levels of liver fibrosis in mice. Silencing of JAK1/2 down-regulated their downstream signaling and inhibited proliferation, migration, and activation of HSCs in vitro, while Ruxolitinib had similar effects on HSCs. Importantly, Ruxolitinib significantly attenuated fibrosis progression, improved cell damage, and accelerated fibrosis reversal in the liver of mice treated with CCl4 or TAA.ConclusionsJAK1/2 regulates the function of HSCs and plays an essential role in liver fibrosis and HCC development. Its inhibitor, Ruxolitinib, may be an effective drug for preventing and treating liver fibrosis.

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.

Ginsenoside Re Improves Inflammation and Fibrosis in Hepatic Tissue by Upregulating PPARγ Expression and Inhibiting Oxidative Stress in db/db Mice.

Ginsenoside Re (Re) is the main component of “Zhenyuan Capsule” (ZYC), which was wildly used in clinic in China for adjunctive treatment of coronary heart disease (CHD) and type II diabetes (T2DM). Nonalcoholic fatty liver disease (NAFLD) is one of the most important complications of T2DM, as well as an important risk factor of CHD. The aim of the present study was to investigate the effects of Re on NAFLD in db/db mice, one of the most recognized gene deficient animal models on T2DM. Sixteen db/db mice and sixteen wild-type mice were divided into four groups and orally administered Re or placebo in equal volume. According to the results, Re showed no obvious effect on blood glucose, lipids, or body weight of db/db mice. Histology pictures of hepatic tissue showed that Re did not improve steatosis, too. However, some evidence suggested that hepatic injury in db/db mice was attenuated by Re administering. Collagen deposition and aminotransferase elevation were significantly downregulated in the DB + Re group compared to those in the DB Group. The mechanisms of the protect effects of Re represented in db/db mice with NAFLD might be inhibiting oxidative stress and the reupregulation of peroxisome proliferator-activated receptor γ (pparγ) expression. The results of this study indicated that ZYC might be able to help T2DM patients with NAFLD to control the progress of NAFLD as an alternation of thiazolidinediones, synthetic agonists of PPARγ, whose side effects and adverse events should not be ignored.

Elevated miR-129-5p attenuates hepatic fibrosis through the NF-κB signaling pathway via PEG3 in a carbon CCl4 rat model.

Hepatic fibrosis is a reversible scaring response to chronic liver injury. MicroRNA (miR)-129-5p might regulate fibrosis-related gene expression. This study is performed to decipher, potential of miR-129-5p to influence the progression of hepatic fibrosis in a carbon tetrachloride (CCl4) rat model. Rat hepatic fibrosis was successfully established by subcutaneous injection of 50% CCl4. RT-qPCR revealed that miR-129-5p was poorly expressed and PEG3 was highly expressed in hepatic fibrosis tissues. As reflected by dual-luciferase reporter gene assay, miR-129-5p targeted and reduced the expression of PEG3. Thereafter, miR-129-5p antagomir or short hairpin RNA against paternally expressed gene 3 (PEG3) was adopted for gain- and loss-of-function assay to determine the molecular regulatory mechanism of miR-129-5p. Moreover, we detected the expression of nuclear factor kappa B (NF-κB) signaling pathway-related proteins and apoptosis-related factors, and made a serological analysis of the rat serum samples. Results showed that upregulated miR-129-5p or downregulated PEG3 led to reduction of the histological changes of liver cirrhosis and lowered the apoptosis rate, via downstream effects on the NF-κB signaling pathway. Thus, the hepatic fibrosis induced by CCl4 can be rescued by upregulated miR-129-5p or downregulated PEG3 expression.

Hypoxia aggravates non-alcoholic fatty liver disease in presence of high fat choline deficient diet: A pilot study

AimNAFLD is a chronic and progressive disease for which there are no FDA-approved drugs available in the market. Drug discovery is a time-consuming procedure and requires screening of hundreds of small molecules to find new chemical entities (NECs) for a particular disease. Current preclinical NAFLD animal models take a longer time, which enhances the duration and expenses of the screening procedure. Hence to shorten the duration, we have proposed a preclinical animal model for rapid induction of non-alcoholic steatohepatitis (NASH), an advanced stage of NAFLD in rats. MethodologyThe animals were divided into three groups; control, high fat choline deficient (HFCD) and high fat choline deficient diet with sodium nitrite (40 mg/kg b.w. i.p. per day) (HFCD + NaNO2) respectively. Four weeks later physical and serum biochemical parameters were assessed, intraperitoneal glucose tolerance test was performed, and histopathology and gene expression were analysed. Key findingsHypoxic stress aggravates the lipid accumulation, ballooning, lobular inflammation and fibrosis in hepatic tissue in presence of HFCD diet. SignificanceThis novel rodent model could be a useful NAFLD model to screen small molecules rapidly for treatment of NASH.

LEFTY2 alleviates hepatic stellate cell activation and liver fibrosis by regulating the TGF-β1/Smad3 pathway

Activated hepatic stellate cells (HSCs) are the major cell type involved in the deposition of extracellular matrix (ECM) during the development of hepatic fibrosis. In this study, we revealed that left-right determination factor 2 (LEFTY2), one of the proteins belonging to the transforming growth factor-β (TGF-β) protein superfamily, was remarkedly decreased in human hepatic fibrosis tissues and in a carbon tetrachloride (CCl4)-induced liver fibrosis mouse model. In addition, TGF-β1 treatment markedly reduced the level of LEFTY2 in HSCs. Importantly, overexpression of LEFTY2 suppressed the activation and proliferation of HSCs. LEFTY2 inhibited the expression of TGF-β1-induced fibrosis-associated genes (α-SMA and COL1a1) in human (LX-2) and rat (HSC-T6) HSC cell lines in vitro. Mechanistically, we demonstrated, for the first time, the role of LEFTY2 in inhibiting TGF-β1/Smad3 signaling, suggesting that there is a mutual antagonism between LEFTY2 and TGF-β1/Smad3 signaling during liver fibrosis. Similarly, we observed that LEFTY2 has a negative effect on its downstream genes, including c-MYC, CDK4, and cyclin D1, in liver fibrosis. Collectively, our data strongly indicated that LEFTY2 plays an important role in controlling the proliferation and activation of HSCs in the progression of liver fibrosis and this could be a potential therapeutic target for its treatment.

Mitochondrial-Targeted and Near-Infrared Fluorescence Probe for Bioimaging and Evaluating Monoamine Oxidase A Activity in Hepatic Fibrosis.

Monoamine oxidase A (MAO-A) is a promising diagnostic marker for cancer, depression, Parkinson's disease, and liver disease. The fluorescence detection of MAO-A in living animals is of extreme importance for the early diagnosis of related diseases. However, the development of specific and mitochondrial-targeted and near-infrared (NIR) fluorescence MAO-A probes is still inadequate. Here, we designed and synthesized four NIR fluorescence probes containing a dihydroxanthene (DH) skeleton to detect MAO-A in complex biological systems. The specificity of our representative probe DHMP2 displays a 31-fold fluorescence turn-on in vitro, and it can effectively accumulate in the mitochondria and specifically detect the endogenous MAO-A concentrations in PC-3 and SH-SY5Y cell lines. Furthermore, the probe DHMP2 can be used to visualize the endogenous MAO-A activity in zebrafish and tumor-bearing mice. More importantly, it is the first time that the MAO-A activity of hepatic fibrosis tissues is detected through the probe DHMP2. The present study shows that the synthesized DHMP2 might serve as a potential tool for monitoring MAO-A activity in vivo and diagnosing related diseases.

CAPN2 acts as an indicator of hepatitis B virus to induce hepatic fibrosis.

This study is aimed to investigate whether calpain 2 (CAPN2) serves as an indicator of the hepatitis B virus (HBV) to induce hepatic fibrosis. Differentially-expressed genes (DEGs) in HBV-induced hepatic fibrosis and normal liver tissues were analyzed, and signal pathway which was analyzed by the Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis using DEGs. Next, the gene-related network map was constructed using the Search Tool for the Retrieval of Interacting Genes. Moreover, CAPN2 protein expression, level of hepatic fibrosis, CAPN2 messenger RNA level, and protein levels of CAPN2, a-SAM, COL3A1, COL1A1, and MAPK1 were determined using Immunohistochemistry (IHC), hematoxylin and eosin, RT-qPCR, and western blot (WB), respectively. There were 420 DEGs screened in HBV-induced hepatic fibrosis and normal liver tissues, among which, 373 were significantly upregulated and 47 were obviously downregulated. KEGG analysis showed that the upregulated DEGs were mainly concentrated in extracellular matrix-receptor interaction, protein digestion, and absorption signaling pathways. The network diagram analysis showed that the DEGs, such as CAPN2, ITGAV, and CCR2, play the key role in the DEG network map, and CAPN2 related to hepatic fibrosis via MAPK1. The increased CAPN2 expression and obvious hepatic fibrosis was displayed in the HBV-induced hepatic fibrosis tissues. In addition, HBV could induce CAPN2 expression, and the interference of CAPN2 could inhibit the expression of hepatic fibrosis markers, including a-SAM, COL3A1, COL1A1, and MAPK1. CAPN2 is regarded as a biomarker of hepatic fibrosis induced by HBV.

Human bone marrow mesenchymal stem cells-derived exosomes alleviate liver fibrosis through the Wnt/\u03b2-catenin pathway

BackgroundMesenchymal stem cells (MSCs) are increasingly being applied as a therapy for liver fibrosis. Exosomes possess similar functions to their parent cells; however, they are safe and effective cell-free reagents with controllable and predictable outcomes. In this study, we investigated the therapeutic potential and underlying molecular mechanism for human bone mesenchymal stem cells-derived exosomes (hBM-MSCs-Ex) in the treatment of liver fibrosis.MethodsWe established an 8-week CCl4-induced rat liver fibrosis model, after which, we administered hBM-MSCs-Ex in vivo for 4 weeks. The resulting histopathology, liver function, and inflammatory cytokines were analyzed. In addition, we investigated the anti-fibrotic mechanism of hBM-MSCs-Ex in hepatic stellate cells (HSCs) and liver fibrosis tissue, by western blotting for the expression of Wnt/β-catenin signaling pathway-related genes.ResultsIn vivo administration of hBM-MSCs-Ex effectively alleviated liver fibrosis, including a reduction in collagen accumulation, enhanced liver functionality, inhibition of inflammation, and increased hepatocyte regeneration. Moreover, based on measurement of the collagen area, Ishak fibrosis score, MDA levels, IL-1, and IL-6, the therapeutic effect of hBM-MSCs-Ex against liver fibrosis was significantly greater than that of hBM-MSCs. In addition, we found that hBM-MSCs-Ex inhibited the expression of Wnt/β-catenin pathway components (PPARγ, Wnt3a, Wnt10b, β-catenin, WISP1, Cyclin D1), α-SMA, and Collagen I, in both HSCs and liver fibrosis tissue.ConclusionsThese results suggest that hBM-MSCs-Ex treatment could ameliorate CCl4-induced liver fibrosis via inhibition of HSC activation through the Wnt/β-catenin pathway.

LncRNA GAS5 restrains CCl4-induced hepatic fibrosis by targeting miR-23a through the PTEN/PI3K/Akt signaling pathway.

Hepatic fibrosis is chronic liver damage with many causes that has a relatively high death rate. The current study showed that long noncoding RNA (lncRNA) growth arrest-specific transcript 5 (GAS5), microRNA-23a (miR-23a), and phosphatase and tensin homolog (PTEN) play important roles in the pathological process of hepatic fibrosis but have a relatively unclear regulatory mechanism. This study aimed to investigate the roles of lncRNA GAS5, miR-23a, and PTEN in the pathological process of hepatic fibrosis and hepatic stellate cell (HSC) activation. We used carbon tetrachloride (CCl4) intraperitoneal injections to establish a rat hepatic fibrosis model and exogenous transforming growth factor-β1 to establish an HSC activation model. Quantitative RT-PCR, Western blot, dual-luciferase reporter system, and RNA pull-down assays were used to investigate which microRNAs and lncRNAs participate in the process of hepatic fibrosis and HSC activation. miR-23a expression increased significantly in hepatic fibrosis tissues and activated HSCs. miR-23a interaction with and degradation of PTEN further influenced the downstream signaling pathway phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin/Snail (PI3K/Akt/mTOR/Snail), causing E-cadherin expression levels to decrease and α-smooth muscle actin and collagen I expression levels to increase. lncRNA GAS5 can be used as a sponge platform for miR-23a to decrease miR-23a expression levels competitively. We revealed the role of the lncRNA GAS5/miR-23a/PTEN/PI3K/Akt/mTOR/Snail signaling pathway in hepatic fibrosis, providing molecular targets for the treatment of hepatic fibrosis. NEW & NOTEWORTHY This is the first study revealing that microRNA-23a (miR-23a) promotes hepatic fibrosis through the phosphatase and tensin homolog/phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin/Snail signaling pathway, and long noncoding RNA (lncRNA) growth arrest-specific transcript 5 (GAS5) can act as a sponge platform for miR-23a. Therefore, lncRNA GAS5/miR-23a may bring molecular targets for hepatic fibrosis therapy.

The pathogenetic role of large intestine dysbiosis in the pathogenesis of the mutually burdened non-alcoholic fatty liver disease and chronic kidney disease

Objective — to establish the probable influence of the microbial state of the colon cavity (MSCC) content on the degree of steatosis and liver fibrosis in patients with non-alcoholic steatohepatitis (NASH) with obesity, depending on the presence of comorbid chronic kidney disease (CKD) and its stages.Materials and methods. The study involved 168 patients with NASH, aged 42 to 55 years examined. All patients were allocated to the following groups: Group 1 consisted of 68 patients with NASH with concomitant obesity of 1st degree, Group 2 consisted of 100 patients with NASH with obesity of 1st degree and a comorbid CKD I—III stages (chronic pyelonephritis). The control groups consisted of 30 practically healthy persons (PHPs), which by age and sex were not statistically significantly different from the main group and the comparison group. Microbiocenosis of MSCC was investigated with microbiological method by sowing ten-fold dilutions of feces on differential-diagnostic nutrient media in accordance with the methodological recommendations «Microbiological diagnosis of dysbiosis» of the Ministry of Health of the USSR (1986). The main markers allowed to make conclusions about dysbiotic changes were: the type of belonging of aerobes and anaerobes, quantitative characteristic (concentration) and the frequency of growth of sown colonies. Verification of the severity of dysbiosis was carried out on the basis of the classification of I.B. Kuvaevo, K.S. Ladodo (1991).Results and discussion. The study revealed changes in the microbial state of the colon cavity (MSCC) content during the comorbid course of NASH with obesity and CKD I—III stages, characterized by the development of deep dysbiosis (II—III stages). With the appearance and prevalence of pathogenic microflora, an increase in the number of opportunistic bacteria and yeast fungi of the genus Candida, a probable deficiency of representatives of normal microbiota: lactobacilli, bifidobacteria, bacteroids. The degree of dysbiosis of MSCC correlates with the content of increases with the growth of the CKD stage, the activity of cytolysis of hepatocytes, endotoxicosis, oxidative and nitrosative stress, degree of hepatocyte steatosis and hepatic tissue fibrosis.Conclusions. An important component of the pathogenesis of non-alcoholic steatohepatitis in obese patients and CKD is metabolic intoxication, which arises as a result of a significant violation of the quantitative and qualitative composition of the microflora of MSCC with the development of deep dysbiosis (II—III stages). The degree of dysbiosis of the colon in patients with a combined course of NASH, obesity and CKD increases with the growth of the CKD stage, the degree of obesity, the activity of cytolysis of hepatocytes, endotoxicosis, oxidative and nitrosative stress, interconnected with the degree of hepatocyte steatosis and hepatic tissue fibrosis.

Knockdown of Fstl1 attenuates hepatic stellate cell activation through the TGF‑β1/Smad3 signaling pathway.

Follistatin‑like 1 (Fstl1) is a secreted glycoprotein that belongs to the follistatin and SPARC (secreted protein, acidic and rich in cysteine) families and was identified to serve a critical role in lung fibrosis. However, the role of Fstl1 in liver fibrosis remains undefined. Therefore, the aim of the present study was to investigate the role of Fstl1 in liver fibrosis. The results indicated that Fstl1 was highly expressed in human hepatic fibrosis tissues and activated hepatic stellate cells (HSCs). Furthermore, knockdown of Fstl1effectively suppressed HSC proliferation and the protein expression levels of α‑SMA and collagen I in transforming growth factor (TGF)‑β1‑treated HSCs. Mechanistically, knockdown of Fstl1 remarkably decreased the phosphorylation level of Smad3 in TGF‑β1‑induced HSCs. Taken together, the present study demonstrated that Fstl1serves an important role in liver fibrosis and target deletion of Fstl1 attenuated HSCs activation through suppressing TGF‑β1/Smad3 signaling pathway. Therefore, Fstl1 may be a potential therapeutic target for the treatment of liver fibrosis.

ЗНАЧЕНИЕ ПОКАЗАТЕЛЕЙ ИММУНОСУПРЕССИИ И ЭЛАСТОМЕТРИИ ПЕЧЕНИ В ТАКТИКЕ ЛЕЧЕНИЯ КО-ИНФЕКЦИИ ВИЧ/HCV

Objective: Correction of the main treatment regimen for HIV/HCV co-infection, taking into account the degree of immunosuppression based on indices of CD4+ cells in 1 μl of blood and the severity of hepatic fibrosis tissue according to the elastometry of the liver Methods: A complex examination of 21 patients with HIV/HCV co-infection was carried out, according to anamnestic data who are the users of long-term injecting drugs. The diagnosis of HIV infection was confirmed by the positive result of the rapid test, ELISA, (enzyme-linked immunosorbent assay), immunoblotting. Hepatitis C virus is diagnosed by detection by ELISA of serum anti-HCV. To determine the degree of liver fibrosis, the method of elastometry on the FibroScan apparatus (France) was used on the basis of the Institute of Gastroenterology. The results were evaluated on a scale of the degree of fibrosis of liver METAVIR from F0 (healthy liver) to F4 (hepatic cirrhosis). The number of CD4+ lymphocytes in 1 μl of blood was determined by flow cytometry. Results: With a targeted comprehensive examination of 21 patients with HIV/HCV co-infection, pronounced oscillations in the results of elastometry of the hepatic tissue (from F0 to F4), regardless of the number of CD4+ cells (24 to 809 cells/μl) were found. Conclusion: The results obtained indicate that there is no direct correlation between the degree of immunosuppression in CD4+ indices and the process of formation of hepatic fibrosis in patients with co-infection with HIV/HCV, which is a significant additional diagnostic criterion in choosing the priority of therapy. Keywords: Elastometry, immunosuppression, co-infection, HIV, HCV.

MiR-10a improves hepatic fibrosis by regulating the TGFβl/Smads signal transduction pathway.

The aim of the present study was to examine the expression variation of the mouse hepatic fibrosis tissue transforming growth factor (TGF)-βl/Smads signal transduction pathway and its correlation with progression of hepatic fibrosis. The promotion effect of microRNA (miR)-10a on hepatic fibrosis and its possible mechanism was also assessed. Forty healthy female 8-week-old C57BL6/J mice were randomly divided into the control group (intraperitoneal injection of 5 µl/g normal saline, twice per week for 8 weeks) and the hepatic fibrosis group (intraperitoneal injection of 5 µl/g 10% CCI4 olive oil, twice per week for 8 weeks), with 20 mice per group. RT-PCR was used to test miR-10a expression in cells in the control and hepatic fibrosis groups. Cell culture and transfection of miR-10a mimics were conducted in the two groups and a Cell Counting Kit-8 was used to test the expression of TGF-β1 and Smad7 in hepatic fibroblasts. It was found that in comparison with the control group, miR-10a expression was significantly increased in the hepatic fibrosis group compared with the control group (P<0.05). The expression quantity of miR-10a was significantly increased in the transfection group compared with the control group (P<0.05). A high expression of miR-10a significantly improved TGF-β1 expression and reduced Smad7 expression in the hepatic fibrosis group (P<0.05). In conclusion, miR-10a expression was high in mouse hepatic tissues, transfection of miR-10a mimics significantly promoted the cell proliferation of hepatic fibrosis, and miR-10a improved hepatic fibrosis by regulating the TGF-βl/Smads signal transduction pathway.

Pathological analysis of liver tissue in patients with chronic hepatitis B complicated with pulmonary tuberculosis

Objective To investigate the differences of hepatic pathology between the chronic hepatitis B (CHB) with pulmonary tuberculosis patients and CHB patients. Methods Seventy-nine treatment-naive patients with CHB complicated with pulmonary tuberculosis (co-infection group) were collected from January 2009 to December 2014, and 79 CHB patients were selected randomly during the same period as CHB group. Hepatic tissue inflammation and fibrosis between the two groups were compared according to Ishak scoring system. Comparison between two groups were conducted by t test when the variance was equal and Mann -Whitney U test when the variance was unequal. Categorical data were compared by χ2 test. Results A total of 59 (74.7%) patients in co-infection group had inflammation≥ G2, compared to 59.5% in the CHB group. The difference between the two groups was statistically significant (χ2=4.128, P=0.042). Forty-one (51.9%) patients in co-infection group had fibrosis≥S2, compared with 44.3% in the CHB group. The difference was not statistically significant (χ2=0.913, P=0.339). Ishak scoring system showed that piecemeal necrosis, portal area inflammation score and total score in co-infection group were all significantly higher than those in CHB group (2.45±1.19 vs 2.05±1.28, 2.70±1.22 vs 2.32±1.08, 13.16±6.51 vs 11.22±5.72, respectively) , with all the differences statistically significant (t=2.055, 2.068 and 1.984, respectively; P=0.042, 0.040 and 0.049, respectively). However, the confluent necrosis in co-infection group was 2.60±1.91 compared to 2.13±1.68 in CHB group (Z=1.137, P=0.257), focal (dot) soluble necrosis was 2.35±1.06 versus 2.16±0.86 (Z=-1.148, P=0.251), and fibrosis was 3.03±1.63 versus 2.45±1.53 (Z=1.541, P=0.125). Conclusion The liver damage in co-infection patients is more severe compared with CHB patients. Key words: Hepatitis B, chronic; Tuberculosis, pulmonary; Hepatitis B virus; Mycobacterium tuberculosis; Liver biopsy; Pathology