Effect of oral exposure to trichloroethylene on JMJD3 expression and polarization of M1 Kupffer cells / 环境与职业医学

Background Trichloroethylene (TCE) can enter human body through biological accumulation of polluted water or air, resulting in health hazards. The most commonly involved organs are the liver. Objective To observe potential polarization of M1 Kupffer cells (KCs) in mice liver exposed to TCE orally, and to investigate the relationship between histones lysin demethylase JMJD3 and M1 KCs polarization. Methods A total of 72 SPF BALB/c mice aged 6 to 8 weeks were randomly divided into a blank control group (n=18), a vehicle control group (n=18), a 2.5 mg·mL−1 TCE group (n=18), and a 5.0 mg·mL−1 TCE group (n=18) after adaptive feed for one week. A TCE transoral exposure model was established after eight weeks of administration according to previous research of the research group. In the 2nd, 4th, and 8th weeks, the mice were sacrificed and liver tissue samples were collected. Western blotting was used to detect the expression level of JMJD3 in the liver tissue samples. Immunofluorescence was used to co-locate the macrophage marker F4/80 and the surface marker CD11c of M1 macrophages. Immunohistochemistry was used to detect the expressions of CD16/32, a marker of M1 macrophages, and TNF-α, an inflammatory factor of M1 macrophages in mouse liver. Results In the 2nd, 4th, and 8th weeks, the mice in each group were generally in good condition, and no individual died due to TCE. There was no statistically significant difference in the amount of water consumed by each group, nor in the body weight gain and the liver coefficient of mice at each time point (P>0.05). The results of Western blotting analysis showed that there was no statistically significant difference in JMJD3 protein expression level between the blank control group and the vehicle control group at each time point, the expression levels of JMJD3 protein in the 2.5 mg·mL−1 TCE group and the 5.0 mg·mL−1 TCE group were higher than that in the control group , and the expression level of JMJD3 protein in the 5.0 mg·mL−1 TCE group was higher than that in the 2.5 mg·mL−1 TCE group (P<0.05). The results of immunofluorescence co-localization showed that the expressions of F4/80 and CD11c were low in the blank control group and the vehicle control group, while the expressions of F4/80 and CD11c were increased in the 2.5 mg·mL−1 and the 5.0 mg·mL−1 TCE groups. The results of immunohistochemistry showed that the expressions of CD16/32 and TNF-α in the blank control group and the vehicle control group were low, and there were large deposits in the 2.5 mg·mL−1 TCE group and the 5.0 mg·mL−1 TCE group. Conclusion The polarization of M1 KCs and the expression of proinflammatory factors may be related to an increased expression level of JMJD3 induced by oral TCE exposure.

Role of Kupffer cell polarization in immune tolerance of liver transplantation / 器官移植

With the maturity of surgical techniques, the success rate of liver transplantation has been gradually increased. However, the establishment of long-term immune tolerance after operation still faces multiple challenges. Kupffer cells are tissue-resident macrophages, which could reside in the liver and polarize into different directions following liver transplantation, forming M1 Kupffer cells and M2 Kupffer cells. M1 Kupffer cells have pro-inflammatory function, whereas M2 Kupffer cells possess immunoregulatory function. It contributes to the establishment of immune tolerance by inhibiting the quantity and function of M1 Kupffer cells, or enhancing the quantity and function of M2 Kupffer cells. The polarization of Kupffer cells is regulated by many cytokines and signals, which provides an opportunity for therapies to establish immune tolerance of liver transplantation by interfering Kupffer polarization. In this article, the relationship between Kupffer cell polarization and immune tolerance of liver transplantation, and the mechanism of Kupffer cell polarization were reviewed, aiming to provide reference for establishing immune tolerance of liver transplantation.

Endoplasmic reticulum stress enhances tumor necrosis factor- expression in rat Kupffer cells to trigger hepatic stellate apoptosis cell through TNFR/caspase-8 pathway / 南方医科大学学报

OBJECTIVE@#To investigate the role of endoplasmic reticulum (ER)-stress of Kupffer cells (KCs) and KCs-derived tumor necrosis factor- (TNF-) in medicating apoptosis of hepatic stellate cell (HSC).@*METHODS@#Sixty male SD rats were randomized into control group, model group, ER- stress group, depletion group and KCs block group (=15). The 4 groups of rats were given intraperitoneal injections (twice a week for 8 weeks) of normal saline (2 mg/kg); 40% CCl4 solution (in peanut oil, 2 mg/kg); 40% CCl4 solution (2 mg/kg) and tunicamycin (1 mg/kg); and 40% CCl4 solution (2 mg/kg) and tunicamycin (1 mg/kg) followed by clodronate liposomes (50 mg/kg), respectively. After the treatments, samples of the liver tissue and serum were collected from the rats from the 4 groups to isolate KC cells, which were co-cultured with LX2 cells. In the depletion group, the rats were injected with anti-rat TNFR mAb (0.35 mg/kg) via the portal vein before isolating the KCs. Liver function examination, Eirius red staining, ELISA, immuno- histochemical staining, and RT-PCR were performed to assess the liver function, liver fibrosis, KC phenotypes, expression of the in fl ammatory factors, and the number of active HSC was detected. The isolated KCs were treated with tunicamycin before co-culture with LX2 cells, and ELISA, RT-PCR and Western blot were performed to examine KC phenotypes, in fl ammatory factors, LX2 cell apoptosis and TNFR/caspase8 pathway activity.@*RESULTS@#Compared with the rats in the control group, the rats in the model group had significantly increased ALT and AST levels, Sirius red staining-positive area, and Desmin-positive cells (activated HSCs) ( < 0.05) with significantly lowered number of CD16-positive KCs (M1), and TNF- protein and mRNA expression ( < 0.05). Compared with those in the model group, the rats in ER-stress group showed significantly decreased ALT and AST levels, Sirius red staining positivity and Desmin-positive cells ( < 0.05) and increased number of CD16-positive KCs and TNF- expressions ( < 0.05). In the depletion group, compared with the ER-stress group, the rats had significantly increased ALT and AST levels of, Sirius red staining positivity and Desmin-positive cells ( < 0.05) and reduced CD16- positive KCs and TNF-expressions ( < 0.05). In the cell co-culture experiment, the model group showed significantly reduced TUNEL-positive LX2 cells, CD16-positive cells, and expressions of TNFR1, cleaved caspase- 8 and cleaved caspase- 3 in the KCs ( < 0.05) with increased Desmin-positive LX2 cells ( < 0.05). Compared with the model group, the ER- stress group exhibited significantly increased TUNEL-positive LX2 cells, CD16-positive cells and expressions of TNFR, cleaved caspase-8 and cleaved caspase-3 in the KCs ( < 0.05) and decreased Desmin-positive LX2 cells ( < 0.05). In the depletion group, blocking TNFR resulted in significantly decreased expressions of cleaved caspase-8 and cleaved caspase-3 compared with those in ER- stress group ( < 0.05) although there was no significant changed in TNFR expression.@*CONCLUSIONS@#ER stress of KCs promotes the transformation of KCs towards M1 phenotype and increases the expression of TNF-, which triggers the apoptosis of HSCs through the TNFR/caspase-8 pathway.

Effect of Soothing Gan (Liver) and Invigorating Pi (Spleen) Recipes on TLR4-p38 MAPK Pathway in Kupffer Cells of Non-alcoholic Steatohepatitis Rats / 中国结合医学杂志

OBJECTIVE@#To investigate the mechanism of inflflammatory-mediated toll-like receptor 4 (TLR4)-p38 mitogen-activated protein kinase (p38 MAPK) pathway in Kupffer cells (KCs) of non-alcoholic steatohepatitis (NASH) rats and the intervention effect of soothing Gan (Liver) and invigorating Pi (Spleen) recipes on this pathway.@*METHODS@#After 1 week of acclimatization, 120 Sprague-Dawley male rats were randomly divided into 8 groups using a random number table (n=15 per group): normal group, model group, low-dose Chaihu Shugan Powder (, CHSG) group (3.2 g/kg), high-dose CHSG group (9.6 g/kg), low-dose Shenling Baizhu Powder (, SLBZ) group (10 g/kg), high-dose SLBZ (30 g/kg) group, and low- and highdose integrated recipe (L-IR, H-IR) groups. All rats in the model and treatment groups were fed with a high-fat diet (HFD). The treatments were administrated by gastrogavage once daily and lasted for 26 weeks. The liver tissues were detected with hematoxylin-eosin (HE) and oil red O staining. Levels of liver lipids, serum lipids and transaminases were measured. KCs were isolated from the livers of rats to evaluate the mRNA expressions of TLR4 and p38 MAPK by real-time flfluorescence quantitative polymerase chain reaction, and proteins expressions of TLR4, p-p38 MAPK and p38 MAPK by Western blot. Levels of inflammatory cytokines including tumor necrosis factor α (TNF-α), interleukin (IL)-1 and IL-6 in KCs were measured by enzyme-linked immunosorbent assay.@*RESULTS@#After 26 weeks of HFD feeding, HE and oil red O staining showed that the NASH model rats successfully reproduced typical pathogenesis and histopathological features. Compared with the normal group, the model group exhibited significant increases in body weight, liver weight, liver index, serum levels of total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol, and aspartate aminotransferase as well as TC and TG levels in liver tissues, and significant decrease in serum level of high-density lipoprotein cholesterol (Plt;0.05 or Plt;0.01), while those indices were significantly ameliorated in the H-IR group (Plt;0.05 or Plt;0.01). Higher levels of TNF-α, IL-1 and IL-6 in KCs were observed in the model group compared with the normal group (Plt;0.01). Significant decreases in TNF-α, IL-1 and IL-6 were observed in the H-SLBZ, H-IR and L-IR groups compared with the model group (Plt;0.05 or Plt;0.01). The mRNA expressions of TLR4 and p38 MAPK and protein expressions of TLR4, p38 MAPK and p-p38 MAPK in KCs in the model group were significantly higher than the normal group (Plt;0.01), while those expression levels in the L-IR and H-IR groups were significantly lower than the model group (Plt;0.05 or Plt;0.01).@*CONCLUSION@#Inflflammation in KCs might play an important role in the pathogenesis of NASH in rats. The data demonstrated the importance of TLR4-p38MAPK signaling pathway in KCs for the anti-inflflammatory effect of soothing Gan and invigorating Pi recipes.

The role of hepatic macrophages in nonalcoholic fatty liver disease and nonalcoholic steatohepatitis / 한국실험동물학회지

Nonalcoholic steatohepatitis (NASH) is becoming common chronic liver disease because of the increasing global prevalence of obesity and consequently Nonalcoholic fatty liver disease (NAFLD). However, the mechanism for progression of NAFLD to NASH and then cirrhosis is not completely understood, yet. The triggering of these hepatic diseases is thought from hepatocyte injury caused by over-accumulated lipid toxicity. Injured hepatocytes release damage-associated molecular patterns (DAMPs), which can stimulate the Kupffer cells (KCs), liver-resident macrophages, to release pro-inflammatory cytokines and chemokines, and recruit monocyte-derived macrophages (MDMs). The increased activation of KCs and recruitment of MDMs accelerate the progression of NAFLD to NASH and cirrhosis. Therefore, characterization for activation of hepatic macrophages, both KCs and MDMs, is a baseline to figure out the progression of hepatic diseases. The purpose of this review is to discuss the current understanding of mechanisms of NAFLD and NASH, mainly focusing on characterization and function of hepatic macrophages and suggests the regulators of hepatic macrophages as the therapeutic target in hepatic diseases.

Protective effects of agmatine on severe trauma induced acute hepatic injury / 中华危重病急救医学

Objective To observe the effect of agmatine (AGM) on inflammatory factor in Kupffer cells of liver,and to investigate the protective effects of AGM on severe trauma-induced liver injury in mice and its possible mechanism.Methods Forty-two adult male BALB/c mice were randomly divided into sham group,model group,and AGM treatment group,with 14 mice in each group.The mice model of trauma-hemorrhage was reproduced by hindlimbs fracture combined with 35％ of orbital bleeding.The mice in the sham group were only anesthetized without other treatments.The mice in AGM treatment group were given intraperitoneal injection of 200 mg/kg AGM when limited recovery was performed,and the mice in model group were given the equal amount of normal saline.Seven mice in each group were sacrificed at 12 hours and 24 hours,respectively,after modeling,and blood samples and liver tissue were harvested,and liver Kupffer cells were isolated.Serum alanine aminotransferase (ALT),aspartate transaminase (AST)and lactic dehydrogenase (LDH) were determined with automatic biochemistry analyzer.Hepatic pathological changes were observed with light microscope using hematoxylin and eosin (HE) staining.The levels of tumor necrosis factor-α(TNF-o) and interleukin-6 (IL-6) in serum,hepatic homogenate and Kupffer cell supernatant were determined with enzyme linked immunosorbent assay (ELISA).The mRNA expressions of pro-inflammatory cytokines TNF-α and IL-6 in the Kupffer cell were determined by real-time fluorescent quantitation reverse transcription-polymerase chain reaction (RT-qPCR).Results ① The normal liver tissue structure was found in sham group.At 24 hours after modeling in the model group,the changes in pathobiology were found as following:neutrophil infiltration,hepatocytes swelling,hyperemia,and necrosis,as well as the abnormality of parameters reflecting liver function.AGM could significantly improve the pathological changes in liver tissue caused by severe trauma,and ameliorate the liver function.② There were no significant differences in the levels of TNF-α and IL-6 in serum and hepatic tissue at 12 hours after modeling,and the parameters at 24 hours in model group were higher than those at 12 hours,which were significantly higher than those of the sham group [serum TNF-α (ng/L):80.8±4.7 vs.34.7±4.7,IL-6 (ng/L):104.0±9.0 vs.55.4±3.3;liver TNF-α (ng/mg):405.2± 19.6 vs.57.2±10.0,IL-6 (ng/mg):58.4±7.7 vs.14.3±2.1,all P ＜ 0.01].AGM could effectively reduce the levels of TNF-o and IL-6 in serum and hepatic tissue [serum TNF-α (ng/L):58.2 ± 3.1 vs.80.8 ± 4.7,IL-6 (ng/L):74.1 ± 6.6 vs.104.0± 9.0;liver TNF-α (ng/mg):248.7 ± 22.5 vs.405.2 ± 19.6,IL-6 (ng/mg):22.5 ± 3.1 vs.58.4 ± 7.7,all P ＜ 0.01].③ The levels of TNF-o and IL-6 in Kupffer cells supernatant were significantly higher than those of the sham group,and they were further increased after lipopolysaccharide (LPS) stimulation for 24 hours.AGM could effectively reduce the levels of TNF-α and IL-6 in Kupffer cells [TNF-α (ng/L):256.6 ± 5.6 vs.465.5 ± 5.2,IL-6 (ng/L):1 185.5 ± 64.4 vs.2 018.8 ± 53.2,both P ＜ 0.01],and also decreased the mRNA expressions of TNF-α and IL-6 [TNF-α mRNA (2-△△Ct):7.2±0.4 vs.13.5±0.4,IL-6 mRNA (2-△△Ct):13.2±0.7 vs.21.3 ± 1.6,both P ＜ 0.01].Conclusion Agmatine can reduce trauma-induced acute hepatic injury via suppression of cytokines release in Kupffer cells,and can ameliorate the liver function.

Estudo do papel das células de kupffer na modulação dos citocromos P450 hepáticos por estímulos inflamatórios

É conhecido que processos inflamatórios podem modular a expressão e atividade deenzimas CYPs. Não é claro, entretanto, o modo pelo qual estímulos inflamatórios regulam aexpressão dessas enzimas. Neste trabalho investigamos a hipótese de que as células deKupffer do fígado exerceriam um papel na modulação dos CYPs hepáticos em resposta àinflamação exacerbada ou sepse. O cloreto de gadolínio, é um inibidor seletivo das células deKupffer, conhecido por atenuar o quadro de inflamação exacerbada, quandoadministradopreviamente ao estímulo inflamatório, em diferentes modelos animais. Algunsautores sugeriram que as células de Kupffer atuariam como intermediários na modulação daatividade de CYPs hepáticos desencadeada por estímulos inflamatórios. Há estudos quesugerem que a diminuição da população das células de Kupfferatenua ou elimina a regulaçãodas CYPs hepáticas por estímulos inflamatórios.Além disso, estudos em culturas dehepatócitos in vitro, na ausência de células de Kupffer, tem constatado a regulação negativa daexpressão de CYPs hepáticas após estimulação com LPS. Nessa linha, o objetivo destepresente trabalho é investigar o papel das células de Kupffer na regulação da atividade deenzimas hepáticas de biotransformação de xenobióticos (CYPs) após estimulação inflamatóriacom LPS. Para isso, os níveis séricos de transaminases, e a histopatologia foram empregadospara avaliar o efeito do tratamento com diferentes doses de GdCl3sobre o tecido hepático.Noexperimento principal para investigar o papel das células de Kupffer, os ratos foram alocadosao acaso em quatro grupos. Foram quantificadosmarcadores bioquímicos no soro dos animaispara evidenciar danos ao tecido hepático causados pelos tratamentos e realizado o examehistopatológico...

It is known that inflammatory processes may modulate the expression and activity ofCYP enzymes. The mode by which inflammatory stimuli regulate CYP expression andactivity, however, remains unclear. Kupffer cells are resident macrophages in the liver andthus play an important role in a systemic inflammatory process or in sepsis. Gadoliniumchloride (GdCl3) has been reported to selectively kill an/or inhibit the activity of Kupffercells. Along this line, it has also been described that Gd decreases exacerbated inflammatoryresponses when it is administered prior to inflammatory stimuli in various animal models.Therole of Kupffer cells in the modulation of CYPs activity triggered by inflammatory stimuli,however, is not entirely clear in the literature. There are studies suggesting that a reduction inthe population of Kupffer cells attenuates the down-regulation of hepatic CYPs induced byinflammatory stimuli. However, GdCl3 was also described to decrease liver CYP activityirrespective of whether it depletes or not Kupffer cells. Moreover, in vitro studies showed thatLPS down-regulates the expression of CYP forms in hepatocyte cell lines in culture (in theabsence of Kupffer cells). To investigate whether Kupffer cells play a role in the regulation ofthe activity of liver xenobiotic biotransformation enzymes (CYPs) by inflammatorystimulation with LPS. The activity of transaminases in the blood serum (a marker for liverinjury) was determined and liver histopathology was evaluated in female Wistar rats. In a setof preliminary tests, rats were treated with different doses of GdCl3 or with LPS for selectingdoses and euthanasia time in the main experiment. In the main study experimental groups.Treatment associated liver injury was evaluated by levels of transaminases and alkalinephosphatase in the blood serum and by liver histopathology examination...

Nitric oxide in liver fibrosis: The role of inducible nitric oxide synthase

The inducible form of nitric oxide synthase (iNOS) is expressed in hepatic cells in pathological conditions. Its induction is involved in the development of liver fibrosis, and thus iNOS could be a therapeutic target for liver fibrosis. This review summarizes the role of iNOS in liver fibrosis, focusing on 1) iNOS biology, 2) iNOS-expressing liver cells, 3) iNOS-related therapeutic strategies, and 4) future directions.

Roles of UⅡ/UT system played in innate immune inflammatory signal pathway TLR4-IRF3 in LPS-stimulated primary Kupffer cells / 中国免疫学杂志

Objecitve:To investigate effects of urotensin Ⅱ( UⅡ)/UT system on innate immune inflammatory signal pathway TLR4-IRF3 in the lipopolysaccharide (LPS)-stimulated Kupffer cells (KCs).Methods: Rat KCs were isolated and cultured.Pro-in-flammatory cytokines including IL-6,IFN-βand IFN-γwere assayed by ELISA in culture supernatant of KCs.Cell surface TLR4 were tested with flow cytometry technique.Expression of IRF3 were tested with real-time PCR and Western blot.Results: Significant increases were showed in IL-6, IFN-βand IFN-γsecretion, TLR4-expressed positive rates and IRF3 mRNA levels in KCs after stimulated by LPS,but were inhibited via urantide pretreatment.In addition,LPS induced upregulation of nuclear IRF3 protein and downregulation of cytoplasm IRF3 protein in KCs,which were blocked by urantide pretreatment.Conclusion:UⅡ/UT system mediates immune inflammatory response in part through activating TLR 4-IRF3 pathway in LPS-stimulated KCs.

Effects of UⅡ/UT system on the expression of inflammatory signal molecules p38 MAPK and NF-κB in LPS-stimulated Kupffer cells / 中华微生物学和免疫学杂志

Objective To investigate the effects of urotensin Ⅱ/urotensin Ⅱreceptor ( UⅡ/UT) system on the expression of inflammatory signal molecules p 38 mitogen-activated protein kinase ( p38 MAPK) and nuclear factor-κB ( NF-κB ) in lipopolysaccharide ( LPS )-stimulated Kupffer cells ( KCs ) . Methods Rat KCs were isolated and purified by means of in situ perfusion and density gradient centrifuga-tion.The isolated cells were randomly divided into six treatment groups including group 1:UⅡ(-) urantide (-)LPS(-), group 2:UⅡ(+)urantide(-)LPS(-), group 3: UⅡ(-)urantide(+)LPS(-), group 4:UⅡ(-)urantide(-)LPS(+), group 5:UⅡ(+) urantide(-) LPS(+) and group 6:UⅡ(-)urantide(+) LPS(+) .Western blot assay was performed to detect p 38 MAPK/p-p38 MAPK protein and NF-κB p65 sub-unit.The DNA-binding activity of NF-κB was tested by electrophoretic mobility shift assay (EMSA).Re-sults There was no significant difference with the expression of p 38 MAPK protein in KCs among the six groups (P>0.05).The expression of p65 protein and p-p38 MAPK and the DNA-binding activity of NF-κB were significantly enhanced in LPS-stimulated KCs from groups 4, 5 and 6 in comparison with those in group 1 (P0.05), but that were decreased in group 6 than those in group 4 (all P<0.01).Conclusion UⅡ/UT system participated in the activation of p38 MAPK and NF-κB signaling pathways in LPS-stimulated primary Kupffer cells .

The role of gut-liver axis in the pathogenesis of liver cirrhosis and portal hypertension

Because of the anatomical position and its unique vascular system, the liver is susceptible to the exposure to the microbial products from the gut. Although large amount of microbes colonize in the gut, translocation of the microbes or microbial products into the liver and systemic circulation is prevented by gut epithelial barrier function and cleansing and detoxifying functions of the liver in healthy subjects. However, when the intestinal barrier function is disrupted, large amount of bacterial products can enter into the liver and systemic circulation and induce inflammation through their receptors. Nowadays, there have been various reports suggesting the role of gut flora and bacterial translocation in the pathogenesis of chronic liver disease and portal hypertension. This review summarizes the current knowledge about bacterial translocation and its contribution to the pathogenesis of chronic liver diseases and portal hypertension.

Inhibition of allogeneic T-cell responses by Kupffer cells expressing indoleamine 2,3-dioxygenase in vitro / 中华肝胆外科杂志

Objective To investigate kupffer cells (KCs) expressing indoleamine 2,3-dioxygenase(IDO)in the inhibition of allogeneic T-cell proliferation in vitro. Methods Real-time PCR was used to investigate the expression of IDO mRNA and FasL mRNA in KCs pretreated with or without IFNγ. High performance liquid chromatography was used to analyze the catabolism of tryptophan by IDO from KCs. Allogeneic T-cell response was used to confirm the inhibition of KCs in vitro. The proliferation of lymphocytes was detected using [3 H] thymidine incorporation. Cell cycle and lymphocyte apoptosis were evaluated by flow cytometric assay. Results Real-time PCR revealed IDO mRNA and FasL mRNA expression in KCs pretreated with IFN-γ. IDO catabolic effect was confirmed by a decrease in tryptophan and increase in kynurenine concentration. KCs expressing IDO and FasL from BABL/c mice acquire the ability to suppress the proliferation of T-cells from C57BL/6, which could be blocked by the addition of 1-methyl-tryptophan and anti-FasL antibody. The co-cultured T-cells with KCs expressing IDO and FasL could induce allogeneic T-cell apoptosis and exhibited cell-cycle arrest in G1. Conclusion In addition to the Fas/FasL pathway, IDO may also play an important role in KCs to inhibit allogeneic T-cell proliferation in vitro.

Effect of cytokines secreted from Kupffer cell on HSC proliferation, apoptosis in hepatic fibrosis process / 中国药理学通报

Liver fibrosis can be caused by chronic liver injury arising from various etiological factors and it is a reversible process.The activation of the hepatic stellate cell(HSC)is the central event in liver fibrosis,since we know that cytokines secreted from Kupffer cell participate in HSC proliferation,apoptosis and ECM metabolism.In this paper we focus on the relationship between HSCs,Kupffer cell,cytokines and the course of hepatic fibrosis.Elucidating this relationship will benefit research on the role of Kupffer and HSCs in hepatic fibrosis.

Research progress on Kupffer cells / 解剖学报

Kupffer cell is a member of the liver nonparenchymal cells, it participates in a variety of physiological activities through phagocytosis, secreting cytokines, antigen-presenting pathways, and is closely related to a variety of diseases such as liver cancer, liver fibrosis, liver injury. This paper presents the progress of research on physiological functions of kupffer cells and its relations with liver.

Effect of Kupffer cells on CD4~+ CD25~+ T cells in schistosome granuloma / 中国血吸虫病防治杂志

Objective To explore the effect of targeting Kupffer ceHs on CD4~+ CD25~+T cells in schistosome gramtloma.Methods A total of 30 six-to eight-week-old C57BL/6 female mice were divided into three groups equally,namely a control group,an infection group with S.japonicum cercariae(10 cercariae per mouse) and an infection group injected with GdCl_3 through the penile vein(15 mg/kg)twice perweek.After8 weeks of theinfection,the number of CD4~+ CD25~+T cells was detected by using flow cytometry and the number of Foxp3 was detected by using immunohistochemistry.For the detection of murine IL-4,IL-5,IL-10,TGF-β1 and IFN-γ,a DuoSet ELISA development kit was used.Results The number of CD4~+ CD25~+T ceils and the level of IL-10 decreased significantly in the infection group injected with GdCl_3 compared with the infection group.GdCl_3 treatment decreased Foxp3 production and the level of ALT,and reduced the inflammatory response in schistosome Granuloma.Conclusion Kupffer ceils Can regulate the response of CD4~+ CD25~+T cells in schistosome granuloma.

Role and mechanism of human Kupffer cells in liver allo-geneic transplantation / 中华肝胆外科杂志

Objective To explore the possible role and mechanism of the Kupffer cells (KCs) in liver allo-geneic transplantation at the early stage. Methods In vitro cell contact coculture system was established. Culture supernatants were collected respectively on the 1st, 2nd, 4th, 6th d after cocul-ture and the KCs and PBMCs were harvested on the 6th day after culture. The expression of HLA-G on the membrane of the KCs and PBMCs was detected with immunochemistry. Nitrate reduction test was used to determine the concentration of nitric oxide. IFN-γ, IL-10, TGF-β1 cytokine levels in the supernatants were also measured with ELISA. The proliferation of lymphocytes was evaluated with MTT. Results six days later, no HLA-G molecules were detected on the membrane of the KCs and PBMCs. In the experimental group containing KCs, the levels of NO, IL-10 and TGF-β1 was signifi-cantly increased(P<0. 05), while the levels of IFN-γ was relatively lower(P<0. 05) as compared to the experimental group without KCs. No IL-10 and IFN-γ were detected in the control group, and on-ly few NO and TGF-β1 was found in the control group with KCs. MTT test showed that the value of optical density was lower in the experimental group with KCs than that in any other group(P<0. 05).Conclusion No HLA-G is expressed on the membrane of KCs and PBMCs after contact coculture.KCs may participate in regulating production of NO and Th2/Th3-like cytokines and suppressing the proliferation of lymphocytes, through which KCs probably take part in inducing immunotolerance of liver transplantation in early stage.

Lipopolysaccharide-induced liver injury expression Kupffer cell and protection mechanisms of S-adenosylmethionine / 国际外科学杂志

S-adenosylmethionine(SAM) is a physiologically active molecule in all the organizations and the human body fluids, and it is important to participate in a variety of biochemical reactions, the regulation of liver regeneration, liver cells differentiation and various sensitivity of the injury. The SAM affects the liver through a variety of ways. It is confirmed that the SAM and MTA can block the LPS-induced TNF-α of Kupffer cells to protect the liver. In a word, S-adenosylmethionine may be beneficial to LPS-induced liver in-jury in clinical treatment.

Effects of CTGF on Kupffer cells induced activation of hepatic stellate cells / 中华普通外科杂志

Objective To investigate the effect of RNA interference connective tissue growth factor (CTGF)expression on Kulclffer cells(KC)induced activation of hepatic stellate cells(HSC).Methods Rat CTGF RNA interference vector Psilencer 3.1H1-Neo-CTGF was constructed and identified.HSC cell line rHSC-99 cells were divided into three groups,group A served as control,group B transfected with vector without CTGF interference.group C was RNA interferenee CTGF expression of HSC.RT-PCR was used to measure the expression of CTGF in HSC.Rat Kur)ffer cells were isolated and identified.and cocultured with HSC in the 3 groups respectively.MTT assay was used to evaluate the proliferation of HSC.RT-PCR was used to measure the expression of TGF-β1 and precollagen type I in HSC.Western blot was used to measure the expression of TGF-β1 in HSC.ELISA was used to detect the production of precollagen type I protein.Immunofluorescence was used to detect the expression of ot-smooth muscle actin(α-SMA)in HSC.Resuits After CTGF RNA interference vector transfection.CTGF expression of HSC decreased by 22%(P＜0.01).The yield rate of Kupffer cell was 5×107 and the cell viability exceeded 98%.In the HSC and KC co-culture system.the proliferation and activation of HSC were inhibited while RNA interferenee CTGF of HSC.As compared with control,HSC proliferation decreased by 29%(P＜0.01).Precollagen type I and ot-SMA expression decreased by 38%(P＜0.01).Production of precollagen type I protein in culture medium decreased bv 48%(P＜0.01).Conclusions Blockade CTGF expression of HSC inhibits KC induced activation of HSC.

The Factors Affecting Transplanted Hepatocytes Repopulation in Rats with Liver Fibrosis

PURPOSE: Cell therapy for various diseases has gained wide acceptance. Because most patients with chronic liver failure have mild-to-severe liver cirrhosis, there are many limitations to clinical applications. We analyzed how to increase cell engraftment in rats with liver fibrosis. METHODS: We used analbuminemic SD rats (NARs) as recipients of syngeneic CAG-EGFP SD hepatocytes obtained by the 2 perfusion method. Hepatic fibrosis was induced with thioacetamide in drinking water for 6 weeks in the recipient NARs. NARs were pre-treated with gadolinium, doxorubicin, and gliotoxin before hepatocyte transplantation. We evaluated the degree of cell engraftment by RT-PCR and immunofluorescent staining for GFP and albumin. The transplanted cells were detected by immunostaining for albumin, and serum albumin was also measured. RESULTS: Although detection of GFP by RT-PCR was variable, albumin was detected in all groups 4 wks after hepatocyte transplantation. GFP and albumin were also detected by immunofluorescent staining 1 and 4 wks after cell transplantation. In control rats, albumin production was maximal at 3 wks, and after that it rapidly decreased. In the gadolinium and doxorubicin-treated group, albumin production was increased up to 4 wks. Albumin production in the gadolinium-treated group was superior to that of the doxorubicin-treated group. CONCLUSION: Kupffer cells play the most important role in cell engraftment in hepatic fibrosis. Therefore, perturbation of kupffer cells in hepatic fibrosis is needed to increase cell engraftment.

The Factors Affecting Transplanted Hepatocytes Repopulation in Rats with Liver Fibrosis

PURPOSE: Cell therapy for various diseases has gained wide acceptance. Because most patients with chronic liver failure have mild-to-severe liver cirrhosis, there are many limitations to clinical applications. We analyzed how to increase cell engraftment in rats with liver fibrosis. METHODS: We used analbuminemic SD rats (NARs) as recipients of syngeneic CAG-EGFP SD hepatocytes obtained by the 2 perfusion method. Hepatic fibrosis was induced with thioacetamide in drinking water for 6 weeks in the recipient NARs. NARs were pre-treated with gadolinium, doxorubicin, and gliotoxin before hepatocyte transplantation. We evaluated the degree of cell engraftment by RT-PCR and immunofluorescent staining for GFP and albumin. The transplanted cells were detected by immunostaining for albumin, and serum albumin was also measured. RESULTS: Although detection of GFP by RT-PCR was variable, albumin was detected in all groups 4 wks after hepatocyte transplantation. GFP and albumin were also detected by immunofluorescent staining 1 and 4 wks after cell transplantation. In control rats, albumin production was maximal at 3 wks, and after that it rapidly decreased. In the gadolinium and doxorubicin-treated group, albumin production was increased up to 4 wks. Albumin production in the gadolinium-treated group was superior to that of the doxorubicin-treated group. CONCLUSION: Kupffer cells play the most important role in cell engraftment in hepatic fibrosis. Therefore, perturbation of kupffer cells in hepatic fibrosis is needed to increase cell engraftment.