Pathological Changes In Liver Research Articles

Ammonia exposure causes the disruption of the solute carrier family gene network in pigs

Ammonia is the main harmful gas in livestock houses. However, the toxic mechanism of ammonia is still unclear. Therefore, we examined the effects of ammonia exposure on different tissues of fattening pigs by histological analysis and transcriptome techniques in this study. The results showed that there were varying degrees of pathological changes in liver, kidney, hypothalamus, jejunum, lungs, spleen, heart and trachea of fattening pigs under ammonia exposure. Notably, the extent of damage in liver, kidney, jejunum, lungs, hypothalamus and trachea was more severe than that in heart and spleen. Transcriptome results showed that ammonia exposure caused changes in 349, 335, 340, 229, 120, 578, 407 and 115 differentially expressed genes in liver, kidney, spleen, lung, trachea, hypothalamus, jejunum and heart, respectively. Interestingly, the changes in solute vector (SLC) family genes were found in all 8 tissues, and the verified gene results (SLC11A1, SLC17A7, SLC17A6, SLC6A4, SLC22A7, SLC25A3, SLC28A3, SLC7A2, SLC6A6, SLC38A5, SLC22A12, SLC34A1, SLC26A1, SLC26A6, SLC27A5, SLC22A8 and SLC44A4) were consistent with qRT-PCR results. In conclusion, ammonia exposure can cause pathological changes in many tissues and organs of fattening pigs and changes in the SCL family gene network. Importantly, the SCL family is involved in the toxic mechanism of ammonia. Our findings will provide a new insight for better assessing the mechanism of ammonia toxicity.

Chrysanthemi Flos extract alleviated acetaminophen-induced rat liver injury via inhibiting oxidative stress and apoptosis based on network pharmacology analysis

Context Acetaminophen (APAP) overdose is the leading cause of drug-induced liver injury. Bianliang ziyu, a variety of Chrysanthemum morifolium Ramat. (Asteraceae), has potential hepatoprotective effect. However, the mechanism is not clear yet. Objective To investigate the hepatoprotective activity and mechanism of Bianliang ziyu flower ethanol extract (BZE) on APAP-induced rats based on network pharmacology. Materials and methods Potential pathways of BZE were predicted by network pharmacology. Male Sprague-Dawley rats were pre-treated with BZE (110, 220 and 440 mg/kg, i.g.) for eight days, and then APAP (800 mg/kg, i.g.) was used to induce liver injury. After 24 h, serum and liver were collected for biochemical detection and western blot measurement. Results Network pharmacology indicated that liver-protective effect of BZE was associated with its antioxidant and anti-apoptotic efficacy. APAP-induced liver pathological change was alleviated, and elevated serum AST and ALT were reduced by BZE (440 mg/kg) (from 66.45 to 22.64 U/L and from 59.59 to 17.49 U/L, respectively). BZE (440 mg/kg) reduced the ROS to 65.50%, and upregulated SOD and GSH by 212.92% and 175.38%, respectively. In addition, BZE (440 mg/kg) increased levels of p-AMPK, p-GSK3β, HO-1 and NQO1, ranging from 1.66- to 10.29-fold compared to APAP group, and promoted nuclear translocation of Nrf2. BZE also inhibited apoptosis induced by APAP through the PI3K–Akt pathway and restored the ability of mitochondrial biogenesis. Discussion and conclusions Our study demonstrated that BZE protected rats from APAP-induced liver injury through antioxidant and anti-apoptotic pathways, suggesting BZE could be further developed as a potential liver-protecting agent.

5-Aminolevulinic acid combined with sodium ferrous citrate (5-ALA/SFC) ameliorated liver injury in a murine acute graft-versus-host disease model by reducing inflammation responses through PGC1-α activation.

Acute graft-versus-host disease (aGvHD) remains lethal as a life-threatening complication after allogeneic hematopoietic stem cell transplantation (HSCT). Inflammatory responses play an important role in aGvHD. 5-Aminolevulinic acid combined with sodium ferrous citrate (5-ALA/SFC) has been widely reported to have a major effect on the anti-inflammatory response; however, these effects in aGvHD models have never been reported. In this study, a murine aGvHD model was developed by transferring spleen cells from donor B6/N (H-2kb) mice into recipient B6D2F1 (H-2kb/d) mice. In addition to evaluating manifestations in aGvHD mice, we analyzed the serum ALT/AST levels, liver pathological changes, infiltrating cells and mRNA expression of inflammation-related cytokines and chemokines. 5-ALA/SFC treatment significantly ameliorated liver injury due to aGvHD and decreased the population of liver-infiltrating T cells, resulting in a reduced expression of pro-inflammatory cytokines and chemokines. Furthermore, the mRNA expression proliferator-activated receptor-γcoactivator (PGC-1α) was enhanced, which might explain why 5-ALA/SFC treatment downregulates inflammatory signaling pathways. Our results indicated that 5-ALA/SFC can ameliorate liver injury induced by aGvHD through the activation of PGC-1α and modulation of the liver mRNA expression of inflammatory-related cytokines and chemokines. This may be a novel strategy for treating this disease.

Safety research on treatment of cough reflex disorder after cerebral hemorrhage with crossing nape electroacupuncture in guinea pig

To observe the effect of crossing nape electroacupuncture(EA) on cough count (CCnt), cough incubation period and blood routine, liver function, renal function, myocardial enzymology, and heart, liver and kidney pathological structure in guinea pig with cough reflex disorder after cerebral hemorrhage, so as to evaluate the efficacy and safety of crossing nape EA in the treatment of cough reflex disorder after cerebral hemorrhage. A total of 160 male pure white guinea pigs were randomly divided into normal group, model group, acupuncture group, crossing nape EA group and nape EA group (n=32 in each group). These 5 groups were further divided into 1, 3, 7 and 14 d subgroups(n=8 in each subgroup). The model of cough reflex disorder after cerebral hemorrhage was established by injection of the guinea pigs' autoblood (50 μL) into the basal ganglia region confirmed by Longa's neurologic examination grading system(0-4 points). Acupuncture was applied to bilateral "Fengchi" (GB20) and "Yifeng" (SJ17) in the acupuncture group. In the crossing nape EA group, the positive pole of the needle is connected to SJ17, while the negative pole is connected to the opposite side of GB20(2 Hz, 0.5 mA). In the nape EA group, the positive pole of the needle is connected to SJ17, while the negative pole is connected to the same side of GB20(2 Hz, 0.5 mA). All treatments were applied 20 min each time, once a day. The changes of CCnt and cough incubation period were recorded by Buxco device. The changes of blood routine, liver function, kidney function, myocardial enzyme and other laboratory indexes were monitored by automatic analyzer instrument. The pathological changes of heart, liver and kidney of guinea pigs were observed by transmission electron microscopy. Compared with the normal group, the guinea pigs' neurological deficit scores in the model group were significantly increased at each time point (P<0.01). Compared with the model group, the neurological deficit scores of the acupuncture group, crossing nape EA group and nape EA group at 3 d, 7 d and 14 d all decreased (P<0.01, P<0.05). Following modeling, the CCnt at each time point in the model group was significantly reduced (P<0.01), and the cough latency was significantly prolonged (P<0.01). After interventions, the CCnts were increased and the incubation periods of cough were shortened (P<0.05, P<0.01) at 7 d and 14 d in the 3 intervention groups, and were superior in the crossing nape EA group than those of acupuncture group and nape EA group (P<0.05, P<0.01). The improvement degrees of these 2 indexes in the nape EA group at 14 d were better than those of the acupuncture group (P<0.05). There were no significant differences among the 3 groups in red blood cells counting, hemoglobin, myocardial enzymes, liver and kidney function (P>0.05). Compared with the normal group, the WBC counts in all the time points (except 1 d after operation) were significantly increased(P<0.01) in the model group. In comparison with the model group, the WBC counts at 3 d in crossing nape EA group, and at 7 d and 14 d in the 3 intervention groups decreased (P<0.01, P<0.05). Meanwhile, the WBC counts were lower in the crossing nape EA group at 7 d and 14 d than those of acupuncture group and nape EA group (P<0.05). The results of transmission electron microscopy showed that the histopathological structure of heart, liver and kidney of guinea pigs in each group had no significant changes. Crossing nape EA is safe and effective in the treatment of cough reflex disorder after cerebral hemorrhage in guinea pigs. In addition, guinea pigs will have an inflammatory response after cerebral hemorrhage, and crossing nape EA may have a certain anti-inflammatory effect.

Effect of basic fibroblast growth factor treatment on efficacy of adipose-derived mesenchymal stem cells in liver cirrhosis

Objective: To observe the effect of basic fibroblast growth factor (bFGF) treatment on efficacy of adipose-derived mesenchymal stem cells (ADSCs) in cirrhotic rats. Methods: A rat model of liver cirrhosis was established via intraperitoneal injection of carbon tetrachloride (CCl(4)) for 10 weeks. Thirty SD rats were randomly divided into 3 groups (n = 10): control group served as group A, and 0.5ml of phosphate-buffered saline (PBS) was transfused into the tail vein; ADSCs single-dose transplantation group served as group B, and 1×10(6) ADSCs were transplanted into the tail vein; bFGF-treated ADSCs treatment group served as group C, and 1×10(6) bFGF-treated ADSCs were transplanted through tail vein. Liver function, pathological and cytokine changes, and the in vivo survival transformation condition of the transplanted cells were measured at one week after transplantation. F test and an independent sample t test were used. Results: bFGF treatment had significantly promoted the proliferation, differentiation and overexpression of hepatocyte growth factor (HGF) in ADSCs [ADSCs single: (2 137.16 ± 261.52) pg/ml vs. ADSCs (bFGF): (4 776.23 ± 532.44) pg/ml, P < 0.05]. The bFGF-treated ADSCs treatment group had statistically significant differences in promoting the recovery of liver function [alanine aminotransferase (ALT): ADSCs single (190.8 ± 34.98) vs. ADSCs (bFGF): (117.8 ± 35.81) pg/ml; aspartate aminotransferase (AST): ADSCs single (295.2 ± 33.71) U/L vs. ADSCs (bFGF): (183.8 ± 41.29) U/L, P ​​< 0.05). There was no statistically significant difference in serum albumin levels between the control group, ADSCs single group, and ADSCs (bFGF) group. Compared with the control group, the serum albumin level of ADSCs (bFGF) group was increased significantly (P < 0.05), and the difference between the control group and the ADSCs single transplantation group in improving liver regeneration and reducing liver damage was statistically significant (P < 0.05). Masson trichrome staining showed that the percentage of the liver fibrosis area in the bFGF-treated ADSCs treatment group was 6.78% ± 0.56%, which was significantly higher than that of the control and ADSCs single dose transplantation group (7.96% ± 0.64%) (P < 0.05). Western blot analysis showed that the expression of HGF protein in the bFGF-treated ADSCs treatment group was significantly up-regulated, while the expression of α-smooth muscle actin was significantly down-regulated, and the difference was significant in the ADSCs single transplantation group. A double fluorescent staining method showed that the numbers of stem cells implanted in the liver tissue of the bFGF-treated ADSCs treatment group were higher than that of the ADSCs single transplantation group. In-vitro cell experiments confirmed that bFGF had significantly promoted the overexpression of HGF in ADSCs. Conclusion: bFGF-treated ADSCs transplantation can significantly improve liver function and fibrosis as compared with ADSCs single-dose transplantation in cirrhotic rats.

PM2.5 disrupts thyroid hormone homeostasis through activation of the hypothalamic-pituitary-thyroid (HPT) axis and induction of hepatic transthyretin in female rats 2.5

Fine particulate matter (PM2.5), a ubiquitous environmental pollutant, has been indicated to affect thyroid hormone (TH) homeostasis in women, but the detailed mechanism behind this effect remains unclear. The objective of this study was to evaluate the roles of the hypothalamic-pituitary-thyroid (HPT) axis and hepatic transthyretin in the thyroid-disrupting effects of PM2.5. Sprague Dawley rats were treated with PM2.5 (0, 15 and 30 mg/kg) by passive pulmonary inhalation for 49 days; and recovery experimental group rats were dosed with PM2.5 (30 mg/kg) for 35 days, and no treatment was done during the subsequent 14 days. PM2.5 was handled twice a day by passive pulmonary inhalation throughout the study. After treatment, pathological changes were analyzed by performing haemotoxylin and eosin staining, measuring levels of THs and urine iodine (UI) in serum, plasma, and urine samples using enzyme-linked immunoabsorbent assay, and expression of proteins in the hypothalamus, pituitary, thyroid, and liver tissues of rats were analyzed by immunohistochemistry and Western blotting. The levels of oxidative stress factors, such as reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (Gpx), and nuclear factor-kappa B (NF-κB) in female rats’ plasma were also evaluated by ELISA. The results of these analyses revealed that PM2.5 treatment induced pathologic changes in rat thyroid and liver characterized by increased follicular cavity size and decreased amounts of follicular epithelial cells and fat vacuoles, respectively. Serum levels of triiodothyronine, thyroxine, and thyroid stimulating hormone were significantly decreased, plasma NF-κB level was increased and plasma redox state was unbalanced (enhanced ROS, MDA and Gpx levels; reduced SOD activities) in female rats treated with PM2.5 (P < 0.05). PM2.5 treatment suppressed the biosynthesis and biotransformation of THs by increasing sodium iodide symporter, thyroid transcription factor 1, thyroid transcription factor 2, and paired box 8 protein expression levels (P < 0.05). Additionally, thyroid stimulating hormone receptor and thyroid peroxidase levels were significantly decreased (P < 0.05). Both thyrotropin releasing hormone receptor and thyroid stimulating hormone beta levels were enhanced (P < 0.05). Moreover, transport of THs was inhibited due to reduced protein expression of hepatic transthyretin upon treatment with PM2.5. In summary, PM2.5 treatment could perturb TH homeostasis by affecting TH biosynthesis, biotransformation, and transport, affecting TH receptor levels, and inducing oxidative stress and inflammatory responses. Activation of the HPT axis and altered hepatic transthyretin levels therefore appear to play a crucial role in PM2.5-induced thyroid dysfunction.

Naringenin alleviates nonalcoholic steatohepatitis in middle-aged Apoe−/−mice: role of SIRT1

BackgroundNaringenin is naturally isolated from citrus fruits possessing many pharmacological activities. However, little is known about the effect of naringenin on nonalcoholic steatohepatitis (NASH) in the model of metabolic syndrome. PurposeThe present study is aimed to investigate the effect of naringenin on NASH in 12-mo-old male ApoE−/− mice and its possible underlying mechanism. MethodsIn vivo, 12-mo-old male ApoE−/− mice were administrated with naringenin by intragastric gavage for 12 weeks. At the end of experiment, the blood samples and liver tissues were collected. Metabolic parameters in serum, levels of triglyceride, cholesterol and hydroxyproline, activities of antioxidant enzymes, and content of inflammatory cytokines (TNF-α and IL-6) in liver were examined by corresponding assay kits. Pathological changes in liver were observed by hematoxylin-eosin, oil red O, masson's trichrome, picro-sirius red and senescence β-galactosidase staining. Dihydroethidium was used for detection of reactive oxygen species (ROS). In vitro, AML-12 cells were treated with oleic acid in the presence or absence of naringenin for 24 h. Transfection of SIRT1 siRNA was also conducted in vitro. Lipid accumulation, cellular ROS generation, malondialdehyde content, antioxidant enzyme activities and secretion levels of TNF-α and IL-6 were examined. Both in vivo and in vitro, gene expressions were detected by real-time PCR or western blot. ResultsNaringenin administration improved metabolic parameters, suppressed hepatic steatosis, regulated expression of genes involved in lipid metabolism (FASN, SCD1, PPARα and CPT1α), reduced hepatic fibrosis and cell senescence, inhibited hepatic inflammation as evidenced by the decreased macrophage recruitment and content of TNF-α and IL-6, and reduced hepatic oxidative stress by suppressing ROS generation and normalizing activities of antioxidant enzymes. Notably, naringenin administration increased hepatic SIRT1 protein expression and activity along with the increased deacetylation of liver kinase B1 (LKB1), PGC1α and NF-κB. In vitro study, the benefits of naringenin on lipid accumulation, oxidative stress and inflammation were diminished by SIRT1 siRNA transfection. ConclusionsThese results indicate that naringenin administration may be a potential curative therapy for NASH treatment and the activation of hepatic SIRT1-mediated signaling cascades is involved in its beneficial effects.

Effects of pine pollen wall on gut microbiota and biomarkers in mice with dyslipidemia.

Pinus yunnanensis pollen is rich in various physiological functions. However, whether the pine pollen wall (PW) plays a beneficial role in the body has not been studied. In this work, we have analyzed its effects on the metabolism and gut microbiota of mouse models of dyslipidemia. We found that the intake of pine PW prevents the liver pathologic changes and reduce the concentrations of TNF-α, IL-6, TC, and high-density lipoprotein cholesterol. Moreover, it can regulate bile acid and fat metabolism, SCFAs content, and the structure of the gut microbiota. According to the change of carbohydrate metabolites, we speculated that cellulose should be the main component to play the above beneficial role, and sporopollenin cannot be utilized in the intestine. Therefore, we consider this study of great significance as it gives a description of biological effects of the pine PW and paves the road to its use in health products.

Corallodiscus flabellata B. L. Burtt extract alleviates lipopolysaccharide/D-galactosamine-induced acute liver failure and brain injury by inhibiting oxidative stress, apoptosis, and inflammation.

Objective(s): Corallodiscus flabellata B. L. Burtt (CF) is distributed along liver meridian, with a possible beneficial effect in the progression of acute liver failure. Therefore, the present study investigates the effect of CF extract on rats with acute liver failure.Materials and Methods:Rats were divided into four experimental groups: Control, Lipopolysaccharide (LPS)/D-Galactosamine (D-GalN) (L/D), Wu Ling Powder + L/D (WLP+L/D) and CF + L/D. Animals were gavage for 7 days, after which all animals except the control group were injected intraperitoneally with LPS and D-GalN to induce acute liver failure. Subsequently, the urine was collected for the next 8 hr, and the liver pathological changes were observed. The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), inflammatory factor and oxidative stress-related indicators were measured. The levels of reactive oxygen species (ROS), apoptosis marker in the liver, water content and aquaporin (AQPs) in the brain were detected. The concentration of ions and osmolality of urine and serum were determined.Results:The results show that CF significantly improved the damage of liver and brain tissue, and reversed the changes of serum ALT, AST, inflammatory factor and Cl-. It modulated oxidative stress-related indicators, reduced the content of ROS, apoptosis markers, water content, the level of Cl- ions and osmolality in the urine and the expression of AQP1, and AQP4 in the brain, and increased the urine output. Conclusion:It was found that the CF extract could alleviate the L/D induced acute liver failure by regulating the hepatocyte apoptosis and AQPs expression in the brain.

Correlation of malondialdehyd and glutathione levels with pathological changes of sheep liver infected with hydatid cyst

This study was conducted to resolve the hypothesis that oxidation stress has been accompanied the hydatid cyst infection. For this purpose, 50 hepatic samples of sheep origin have been inspected and results revealed for that hypothesis via the elevation of MDA and reduction in GSH levels of the germinal layers of that hydatid cyst. Furthermore, the results elucidate that oxidative stress status will appear both of the liver infested only with hydatid cyst or accompanied with other pathological lesions namely, coagulative and liquifactive necrosis, fatty changes and liver fibrosis and cirrhosis.

Anti-obesity effect of Lactobacillus plantarum CQPC01 by modulating lipid metabolism in high-fat diet-induced C57BL/6 mice.

Lactic acid bacteria (LAB) are an important group of microorganisms in the food industry. LAB with health benefits are considered natural elements promoting consumer health. The study investigated the anti-obesity effects of Lactobacillus plantarum CQPC01 (LP-CQPC01) on high-fat diet-induced mice. Liver morphology, liver function indexes, lipid indexes, and inflammatory factors in the serum and liver were determined. Expressions of lipid metabolism-related gene were also detected by qPCR. LP-CQPC01 slowed the HFD-induced increase in body weight, decreased the organ indexes, alleviated hepatic lipid accumulation, and inhibited the increased adipose cell volume. LP-CQPC01 decreased lipid levels of serum and liver, and the contents of pro-inflammatory factors, and increased the IL-4 and IL-10 contents. LP-CQPC01 downregulated the expressions of the C/EBP-α and PPARγ mRNA and upregulated CYP7A1, CPT1, LPL, CAT, SOD1, and SOD2 mRNA. Our results indicated that LP-CQPC01 is a potential probiotic for preventing or alleviating high-energy intake-related lipid conditions. PRACTICAL APPLICATIONS: Obesity is a factor of a variety of cardiovascular diseases. Therefore, it is necessary to suppress the occurrence of fat accumulation in time. This study investigated the effect of LP-CQPC01 on lipid regulation in mice fed a high-fat diet (HFD) and clarified the mechanism of strain to alleviate obesity by enhancing the decomposition of cholesterol and detoxification of fat. LP-CQPC01 reduced fat accumulation without oxidative damage, and was confirmed by the attenuated pathological changes of liver. This research can serve as a significant reference for future research, prevention, and treatment of high-energy intake-related lipid conditions, and the development of functional foods with anti-obesity activity.

Liver Pathologic Changes After Direct-Acting Antiviral Agent Therapy and Sustained Virologic Response in the Setting of Chronic Hepatitis C Virus Infection.

Treatment of chronic viral hepatitis C (HCV) infection with direct-acting antiviral agents (DAAs) results in cure, or sustained viral response (SVR), in more than 90% of patients. However, there are subsets of patients who have persistent liver inflammation and fibrosis and develop hepatocellular carcinoma (HCC) despite achieving SVR. A possible reason for these phenomena may be the presence of virus particles in liver tissue but not blood, otherwise defined as occult infection. To describe liver histologic findings following successful DAA therapy, test HCV RNA by (liver) tissue polymerase chain reaction in treated cases, and identify predictive markers for HCC development in treated cases. A total of 96 identified patients were divided into 4 groups, each differentiated by the presence or absence of SVR and HCC. Groups were compared for several clinicopathologic variables, including degree of inflammation and fibrosis, and the 'directionality' of fibrosis in cirrhotic livers using the novel progressive-indeterminate-regressive scoring system. Overall, we found a significant decrease in inflammation in SVR patients. None of the patients showed regression of their cirrhosis following treatment. No evidence of occult HCV infection was seen in 40 livers tested, including 21 with HCC. The number of patients who developed HCC was similar in the SVR and non-SVR groups, and increased inflammation and fibrosis were associated with HCC development. Following DAA-SVR there appears to be an overall decrease in inflammation, but the fibrosis tends to persist, at least in the short term (median follow-up of 20.2 months).

Yttrium-90 Portal Vein Radioembolization in Sprague-Dawley Rats: Dose-Dependent Imaging and Pathological Changes in Normal Liver.

Portal vein embolization (PVE) is an established neoadjuvant method to induce future liver remnant hypertrophy prior to surgical resection of hepatic tumors. The purpose of our study was to examine the feasibility of PVE with glass 90Y microspheres (Y90 PVE) in Sprague-Dawley rats. We tested the hypothesis that increased doses of Y90 PVE would increase target lobe fibrosis and atrophy. Twenty-two rats were assigned to four groups for Y90 PVE to the right median lobe: very high- (273.8MBq; n = 2), high- (99.9MBq; n = 10), medium- (48.1MBq; n = 5), and low-dose (14.8MBq; n = 5). An untreated control group included seven rats. 90Y PET/CT of 90Y distributions confirmed lobar targeting. MRI volumes were measured at baseline, 2-, 4-, 8- and 12-weeks. Explanted hepatic lobes were weighed, sectioned, and stained for H&E and immunohistochemistry. Digitized slides allowed quantitative measurements of fibrosis (20 foci/slide). Ex vivo measurements confirmed 91-97% activity was localized to the target lobe (n = 4). The percent growth of the target lobe relative to baseline was -5.0% (95% CI -17.0-6.9%) for high-, medium dose rats compared to + 18.6% (95% CI + 7.6-29.7%) in the low-dose group at 12-weeks (p = 0.0043). Radiation fibrosis increased in a dose-dependent fashion. Fibrotic area/microsphere was 22,893.5, 14,946.2 ± 2253.3, 15,304.5 ± 4716.6, and 5268.8 ± 2297.2μm2 for very high- (n = 1), high- (n = 4), medium- (n = 3), and low-dose groups (n = 5), respectively. Y90 PVE was feasible in the rat model, resulted in target lobe atrophy, and dose-dependent increases in hepatic fibrosis at 12weeks. The onset of imaging-based volumetric changes was 8-12weeks.

Hepatic dysfunction induced by intestinal dysbacteriosis mainly manifests as immunologic abnormity in mice.

Currently, the potential role of the alterations occurring in the liver immune system and intestinal flora in liver injury remains unknown. Our study aimed to explore the impacts of intestinal microbial barrier damage induced by ceftriaxone on liver immunity. We developed the BALB/c mice model by administering ceftriaxone. The intestinal microbial barrier damage was observed by 16S rRNA, and the pathological changes of intestines and livers were detected by H&E or transmission electron microscope. The activation of immunocytes were tested by Flow Cytometry; the expression of LPS, ALT, AST, IL-6 and TNF-α were detected by Limulus Test or ELISA. Compared to the control, the intestinal microbes significantly decreased in ceftriaxone group. Additionally, the weight of cecum contents increased, the intestinal wall became thinner and the villus in the small intestine and cecum were damaged. The expression of LPS and the ratio of liver lymphocytes were significantly increased. H&E results indicated the structures of liver arose the pathologic changes. Meanwhile, the content of serum ALT, AST, IL-6 and TNF-α increased. Collectively, our study indicates that the damages of gut microbial barrier induced by ceftriaxone prompted activation of immunocytes and release of inflammatory cytokines, which may lead to chronic inflammation in liver.

Study on growth characteristics of Candida auris under different conditions in vitro and its in vivo toxicity

To investigate the characteristics of growth and metabolism and the in vivo toxicity of Candida auris under different conditions. We observed the growth of Candida auris and Candida albicans under routine culture conditions and in different pH and salt concentrations, and compared their activities of sugar fermentation using microbiochemical reaction tubes. Four-week-old nude mice were randomized into Candida auris infection group (n=5), Candida albicans infection group (n=5) and control group (n=5) for intragastric administration of 0.3 mL suspension the two Candida species (5×109 cfu/mL) or 0.3 mL normal saline. Samples of the liver, kidney, intestine, feces and blood were taken for analysis of the in vivo distribution and toxicity of Candida albicans by fungal culture and histopathological examination. Candida auris exhibited logarithmic growth at 8-24 h after inoculation and showed stable growth after 24 h. Candida auris showed optimal growth within the pH value range of 5-7 with a growth pattern identical to that of Candida albicans. Candida auris grew better than Candida albicans in media containing 5% and 10% NaCl, and could ferment glucose, sucrose, trehalose and sorbitol. Candida auris could be isolated from the feces, blood, liver and kidney of infected nude mice, and the liver had the highest fungal load (5.7 log10 cfu/g). Candida auris could cause pathological changes in the liver and intestine of the mice, but with a lesser severity as compared with Candida albicans. Candida auris exhibits optimal growth in mildly acidic or neutral conditions with a high salt tolerance, and can potentially penetrate the intestinal barrier into blood and lead to tissue injuries in hosts with immunosuppression.

Effects of moxibustion or moxa smoke on serum lipids and carotid plaque stability in atherosclerotic mice

Abstract Objective To observe the effects of moxibustion or moxa smoke on serum lipids, aorta and liver pathology, and carotid plaque stability in atherosclerosis. Methods Fifty-four 8-week-old ApoE−/- mice were randomly divided into three groups (untreated, moxibustion, and moxa smoke) and received a high-fat diet. Eighteen wild-type C57BL/6 mice of the same age were used as controls. The intervention (none, moxibustion between the nipples, or 10–15 mg/m3 moxa smoke) was applied to restrained mice 20 min per day, six days per week, for 12 weeks. At the end of the experimental period, we measured serum lipids and apolipoprotein, stained thoracic aortas and livers to observe pathological changes, and used immunohistochemical staining to assess the levels of α-smooth muscle actin, CD68, tumor necrosis factor-α, nuclear transcription factor-κB, and P38 mitogen-activated protein kinase. We also measured the levels of matrix metalloproteinase-2 and 9 and tissue metalloproteinase inhibitor-1. Results After 12 weeks, lipid metabolism disorder and atherosclerotic plaques were observed in the ApoE−/- mice. Moxibustion or moxa smoke reduced the levels of serum total cholesterol, triglycerides, low density lipoprotein, and very low density lipoprotein but did not affect the levels of high density lipoprotein, apolipoprotein A1, or oxidized low density lipoprotein. Moxibustion or moxa smoke suppressed pathological changes in thoracic aortas and livers, increased fiber cap thickness, the fiber cap thickness/intimal medial thickness ratio, and collagen area percentage, and reduced extracellular lipids. Treatment with moxibustion or moxa smoke increased α-smooth muscle actin and reduced CD68 and the vulnerability index, suppressed tumor necrosis factor-α and nuclear transcription factor-κB expression, and did not affect P38 mitogen-activated protein kinase expression. Treatment lowered the levels of matrix metalloproteinase-2 and 9 and increased those of tissue metalloproteinase inhibitor-1. Conclusion Moxibustion or moxa smoke exert protective effects in serum lipid profiles and carotid plaque stability in atherosclerotic mice by regulating plaque stability, inflammatory factors, and matrix metalloproteinases.

Study of the effect of CHOP signaling molecule in PPARα activation and inhibition with response to inflammation in mice with acute liver failure

Objective: To investigate the mechanism of action of peroxisome proliferator-activated receptor α (PPARα)-mediated CCAAT/enhancer binding protein homologous protein (CHOP) signaling molecule with response to inflammation in mice with acute liver failure. Methods: C57BL/6 mice were used as the research subjects, and D-galactose (D-GalN) combined with lipopolysaccharide (LPS) was injected intraperitoneally to establish a mouse model of acute liver failure. PPARα was activated by Wy-14643. CHOP expression was promoted by plasmids. Liver pathological changes and serum transaminases (ALT and AST) were detected in mice to evaluate liver function. The mRNA expression level of inflammatory factors in liver tissue was detected by real-time fluorescence quantitative PCR. LPS-stimulated macrophage was used to establish an inflammation model. PPARα and CHOP expression was inhibited by siRNA. The mRNA expression level of inflammatory factors in the cells was detected by real-time fluorescence quantitative PCR. Results: Promoted PPARα activation had inhibited liver hemorrhage and inflammation in mice with acute liver failure induced by D-GalN/LPS. In addition, the serum level of transaminases and genetic level of inflammatory factors in liver tissues were reduced (P < 0.01). CHOP accelerated expression had reversed the hepatoprotective effect of PPARα activation, aggravated liver injury, and increased inflammatory factors expression (P < 0.01). At the cellular level, the inhibition of PPARα activation had accelerated the increase of inflammatory factors (P < 0.01), while the inhibition of CHOP activation had all over again decreased the inflammatory factors (P < 0.01). Conclusion: PPARα and CHOP are important signaling molecules to regulate the inflammatory response in acute liver failure and liver injury. PPARα acceleration can down-regulate CHOP to inhibit inflammatory factors, which might play a protective role in mice with acute liver failure.

Ligustilide alleviates the insulin resistance, lipid accumulation, and pathological injury with elevated phosphorylated AMPK level in rats with diabetes mellitus

Background Diabetes mellitus (DM) is one of the major risk factors of disability and death worldwide. Despite of the protective role of ligustilide (LIG) in many cell types, we aimed to investigate whether LIG could be a potential to treat DM. Methods Sprague Dawley rats were randomly assigned to five groups. Rats except control were raised on a high-fat diet (HFD). Streptozotocin was intraperitoneally injected into HFD-fed rats to construct DM model. Rats in the LIG intervention groups received intraperitoneal injection of LIG (10, 20, and 40 mg/kg) post-induction of DM. Blood glucose, plasma insulin (p-insulin), adiponectin, HbA1C%, obesity index, HOMA-IR, and biochemical parameters were estimated. Histopathological analysis and apoptosis in liver and kidney, along with proliferation and apoptosis of islet β-cells, were analyzed. Expression of CPT-1 and ACC, and phosphorylation of Nrf2 and AMPKα1, were finally assessed. Results DM-induced alterations were all relived by LIG intervention. In brief, obesity index, glucose level, P-insulin content, HbA1C, and HOMA-IR were lowered while adiponectin level was elevated. Meanwhile, levels of TC, TG, ALT, and AST were decreased in the LIG intervention groups, along with up-regulated CPT-1 level and down-regulated ACC level. Pathological changes in liver and kidney tissues were alleviated, and apoptotic cells were reduced by LIG treatment. For islet β-cells, LIG up-regulated Ki67 and c-Myc expression, and mitigated ratios of Bax/Bcl-2 and cleaved cas3(9)/cas3(9). Finally, LIG could promote phosphorylation of Nrf2 and AMPKα1. Conclusions LIG alleviated the insulin resistance, lipid accumulation, and pathological injury with the activation of AMPK pathway in DM rats.

MTMR14 protects against hepatic ischemia-reperfusion injury through interacting with AKT signaling in vivo and in vitro

Hepatic ischemia-reperfusion (IR) injury is characterized by severe inflammation and cell death. However, very few effective therapies are presently available for hepatic IR injury treatment. Here, we reported a protective function and the underlying mechanism of myotubularin-related protein 14 (MTMR14) during hepatic IR injury. Hepatocyte-specific MTMR14 knockout (HKO) and transgenic (TG) mice were subjected to hepatic IR operation to explore MTMR14 function in vivo. Primary hepatocytes isolated from MTMR14-HKO and MTMR14-TG mice were subjected to hypoxia/reoxygenation (HR) insult in vitro. We found that MTMR14 expression in liver tissues from individuals with hepatic IR was markedly decreased, and similar results were detected in mice with hepatic IR surgery. MTMR14-TG mice following hepatic IR operation had obviously ameliorated liver pathological changes, along with improved hepatic dysfunction, which was proved by the decreased serum alanine amino transferase (ALT) and aspartate amino transferase (AST) levels. MTMR14-HKO and MTMR14-TG animal models indicated that MTMR14 alleviated cell death and inflammatory response. In addition, MTMR14 inhibited nuclear transcription factor κB (NF-κB) signaling. Of note, promoting MTMR14 expression improved phosphatidylinositol 3-kinase/protein kinase-B (PI3K/AKT) pathway through a physical interaction with AKT, subsequently reducing cell death and inflammation. Therefore, MTMR14 is a protective factor during hepatic IR injury, and the MTMR14/AKT signaling is involved the pathogenesis hepatic IR injury. Improvement of this axis might be a novel therapeutic strategy for the prevention of this pathological process.

Cantharidin-induced acute hepatotoxicity: the role of TNF-α, IKK-α, Bcl-2, Bax and caspase3.

Cantharidin is of high medicinal value but has strong toxicity. Nowadays, multiple research has focused on the mechanism of its antitumor activity while research on toxicological profiles associated with cantharidin poisoning is still limited. Its hepatotoxicity has attracted attention recently for the crucial role of the liver in detoxification. Here, we aim to find a potential mechanism for cantharidin-induced acute hepatotoxicity with a view to assisting subsequent research or clinical use or detoxification. Twenty-one male Sprague-Dawley rats were randomly divided into control, low-dose (1.34 mg/kg) and high-dose (2.67 mg/kg) cantharidin exposure groups. We used hematoxylin-eosin to observe pathological changes and used immunofluorescent staining, western blotting and real-time quantitative polymerase chain reaction to detect the expression of the markers. The main pathological changes in livers of cantharidin-treated rats were necrosis, inflammatory infiltration and hemorrhage. We found coexpression of tumor necrosis factor alpha (TNF-α), IkappaB kinase-alpha (IKK-α) and caspase3 by immunofluorescent staining in livers of cantharidin-treated rats. Compared with the control, the levels of TNF-α, IKK-α and caspase3 increased significantly in the experimental groups (P < .05). The ratio of B-cell lymphoma-2 (Bcl-2)/Bax increased in the low-dose group but decreased in the high-dose group (P < .05). Cantharidin exposure raised IKK-α mRNA and caspase3 mRNA levels (P < .05). In conclusion, the participation of TNF-α, IKK-α, Bcl-2, Bax and caspase3 uncovered a novel mechanism underlying cantharidin-induced acute hepatotoxicity, and the mechanism needs to be studied further.