Killer cell Ig-like receptor (KIR) binds to HLA class I molecules on the surface of target cells, and it confers inhibitory signals to NK cells. Although NK cytotoxicity can be affected by the change of the surface expression of KIR on NK cells, the effect of cytokines on the regulation of KIR expression has not been thoroughly investigated. Here in our study, we investigated the effect of several cytokines, including IL-2, TGF-beta, IFN-gamma, IL-12 and IL-18, on the surface expression of CD158 KIR, which binds to HLA-C, by the use of FACS analysis. In the isolated NK cells, IL-2 obviously increased the surface expression of CD158 KIR after 72 hr in vitro culture, and this was evidenced by the increased percentage of CD158+ NK cells and the increased mean fluorescence intensity of CD158 in CD158+ NK cells. In contrast, TGF-beta decreased the surface expression of CD158 KIR after 72 hr culture. However, IFN-gamma, IL-12 and IL-18 did not change the expression of CD158 KIR. The modulated expression of KIR by IL-2 and TGF-beta can be associated with the changed NK-cytotoxic target-discriminating ability of NK cells upon their exposure to IL-2 and TGF-beta.
Antineoplastic Agents/*pharmacology ; Cells, Cultured ; Human ; Interferon Type II/pharmacology ; Interleukin-12/pharmacology ; Interleukin-18/pharmacology ; Interleukin-2/*pharmacology ; Killer Cells/cytology/*drug effects/*metabolism ; Receptors, Immunologic/*metabolism ; Support, Non-U.S. Gov't ; Transforming Growth Factor beta/*pharmacology

Acetylcysteine ; pharmacology ; Adolescent ; Adult ; Aged ; Female ; Hepatitis, Viral, Human ; blood ; Humans ; Interleukin-18 ; blood ; Male ; Middle Aged

OBJECTIVETo determine the efficacy of the plant-derived bioflavonoid, quercetin, for treating nonalcoholic fatty liver disease (NAFLD) by using a rat model, and to investigate the molecular mechanism underlying its therapeutic effects.

METHODSOne-hundred Sprague-Dawley rats were randomly assigned into the normal control group (normal group), untreated NAFLD model control group (model group), 75 mg/kg/day quercetin treatment group (low-dose group), and 300 mg/kg/day quercetin treatment group (high-dose group). The NAFLD rat model was established by providing four weeks of a high-fat diet; the normal group received normal rat chow diet. The quercetin treatments were administered for eight weeks after model establishment and control groups received simultaneous gavages of isotonic saline, with continuation of the respective diets. At the end of the eight weeks (experimental week 12), the rats were sacrificed for liver and serum collection. Intergroup differences in liver index, fasting blood glucose (FBG), triglycerides (TG), interleukin (IL)-18, IL-10, malondialdehyde (MDA), and histopathological features were assessed by independent samples t-test (normal vs. model), one-way ANOVA (model vs. treatments), and least significant difference t-test (pairwise comparisons); correlations were assessed by Pearson's correlation coefficient.

RESULTSCompared with the normal group, the model group showed significantly higher liver index (t=-2.327), FBG (t=-3.482), TG (t=-0.302), and serum IL-18 (t=-2.704) (all P less than 0.05), but significantly lower IL-10 (t=2.622, P less than 0.05); the MDA level was also higher in the model group, but the difference was not significant (t=-1.083, P less than 0.05). Livers from the model group showed obvious histological features of inflammation (lymphocyte and neutrophil infiltration) and steatosis (cytoplasmic lipid droplets). Inflammation was positively correlated with IL-18 (P less than 0.05), but negatively correlated with IL-10 (P less than 0.05), while steatosis was negatively correlated with IL-10 (P less than 0.05). Compared to the model group, quercetin treatment (both low- and high-dose) led to significant decreases in the liver index, FBG and IL-18 (all, P less than 0.01), and significant increase in IL-10 (P less than 0.05); however, the changes in liver index, FBG and IL-10 were not significantly different between the low- and high-dose treatment groups, but the high-dose of quercetin did induce a significantly greater decrease in IL-18 than the low-dose (P less than 0.05).

CONCLUSIONNAFLD rats have higher serum levels of IL-18 but lower levels of IL-10 than their healthy counterparts, and these differential cytokine expressions may be related to liver inflammation and steatosis. Quercetin treatment may help to delay the progression of NAFLD, possibly by adjusting the balance of inflammatory cytokines.

Animals ; Fatty Liver ; blood ; drug therapy ; Interleukin-10 ; blood ; Interleukin-18 ; blood ; Male ; Non-alcoholic Fatty Liver Disease ; Quercetin ; pharmacology ; therapeutic use ; Rats ; Rats, Sprague-Dawley

This study aims to explore the effect of "Trichosanthis Fructus-Allii Macrostemonis" combination(GX) on the activation of NOD-, LRR-, and pyrin domain-containing protein 3(NLRP3) inflammasome, the release of inflammatory cytokines, and the level of autophagy in RAW264.7 macrophage damaged by lipopolysaccharide(LPS), and the mechanism of GX against inflammatory response in macrophages. To be specific, LPS was used to induce the injury of RAW264.7 cells. Cell Counting Kit-8(CCK-8) assay was employed to measure the survival rate of cells, and Western blot to detect the protein expression of NLRP3, apoptosis-associated speck-like protein(ASC), cysteine-aspartic acid protease(caspase)-1, interleukin(IL)-18, IL-1β, microtubule-associated protein light chain 3(LC3)-Ⅱ, and selective autophagy junction protein p62/sequestosome 1 in RAW264.7 macrophages. ELISA was used to measure the levels of IL-18 and IL-1β in RAW264.7 cells. Transmission electron microscopy was applied to observe the number of autophagosomes in RAW264.7 cells. Immunofulourescence staining was used to detect the expression of LC3-Ⅱ and p62 in RAW264.7 cells. The result showed that GX significantly reduced the protein expression of NLRP3, ASC, and caspase-1 in RAW264.7 cells, significantly increased the protein expression of LC3Ⅱ, decreased the expression of p62, significantly inhibited the secretion of IL-18 and IL-1β, significantly increased the number of autophagosomes, significantly enhanced the immunofluorescence of LC3Ⅱ, and reduced the immunofluorescence of p62. Furthermore, 3-methyladenine(3-MA) could reverse the inhibitory effect of GX on NLRP3, ASC, and caspase-1 and reduce the release of IL-18 and IL-1β. In summary, GX can increase of the autophagy activity of RAW264.7 and inhibit the activation of NLRP3 inflammasome, thereby reducing the release of inflammatory cytokines and suppressing inflammatory response in macrophages.
Inflammasomes/metabolism* ; NLR Family, Pyrin Domain-Containing 3 Protein/metabolism* ; Interleukin-18/metabolism* ; Lipopolysaccharides/pharmacology* ; Macrophages ; Cytokines/metabolism* ; Caspase 1/metabolism* ; Autophagy ; Interleukin-1beta/metabolism*

The aim of this paper was to explore the effects of triptolide( TP),the effective component of Tripterygium wilfordii on improving podocyte epithelial-mesenchymal transition( EMT) induced by high glucose( HG),based on the regulative mechanisms of Nod-like receptor protein 3( NLRP 3) inflammasome in the kidney of diabetic kidney disease( DKD). The immortalized podocytes of mice in vitro were divided into the normal( N) group,the HG( HG) group,the low dose of TP( L-TP) group,the high dose of TP( HTP) group and the mannitol( MNT) group,and treated by the different measures,respectively. More specifically,the podocytes in each group were separately treated by D-glucose( DG,5 mmol·L~(-1)) or HG( 30 mmol·L~(-1)) or HG( 30 mmol·L~(-1)) + TP( 5 μg·L~(-1))or HG( 30 mmol·L~(-1)) + TP( 10 μg·L~(-1)) or DG( 5 mmol·L~(-1)) + MNT( 24. 5 mmol·L~(-1)). After the treatment of HG or TP at 24,48 and 72 h,firstly,the activation of podocyte proliferation was investigated. Secondly,the protein expression levels of the epithelial markers in podocytes such as nephrin and ZO-1,the mesenchymal markers such as collagen Ⅰ and fibronectin( FN) were detected,respectively. Finally,the protein expression levels of NLRP3 and apoptosis-associated speck-like protein( ASC) as the key signaling molecules of NLRP3 inflammasome activation,as well as the downstream effector proteins including caspase-1,interleutin( IL)-1β and IL-18 were examined,severally. The results indicated that,for the cultured podocytes in vitro,HG could cause the low protein expression levels of nephrin and ZO-1,induce the high protein expression levels of collagen Ⅰ and FN and trigger podocyte EMT. Also HG could cause the high protein expression levels of NLRP3,ASC,caspase-1,IL-1β and IL-18 and induce NLRP3 inflammasome activation. On the other hand,the co-treatment of TP( L-TP or H-TP) and HG for podocytes could recover the protein expression levels of nephrin and ZO-1,inhibit the protein expression levels of collagen Ⅰ and FN and ameliorate podocyte EMT. Also the co-treatment of TP( L-TP or H-TP) and HG could down-regulate the protein expression levels of NLRP3 and ASC,inhibit NLRP3 inflammasome activation and reduce the protein expression levels of the downstream effector molecules including caspase-1,IL-1β and IL-18. On the whole,HG could activate NLRP3 inflammasome and induce podocyte EMT in vitro. TP at the appropriate dose range could inhibit NLRP3 inflammasome activation and ameliorate podocyte EMT,which may be one of the critical molecular mechanisms of TP protecting againstpodocyte inflammatory injury in DKD.
Animals ; Caspase 1/metabolism* ; Cells, Cultured ; Diabetic Nephropathies ; Diterpenes/pharmacology* ; Epithelial-Mesenchymal Transition ; Epoxy Compounds/pharmacology* ; Glucose ; Inflammasomes/metabolism* ; Interleukin-18/metabolism* ; Interleukin-1beta/metabolism* ; Mice ; NLR Family, Pyrin Domain-Containing 3 Protein/metabolism* ; Phenanthrenes/pharmacology* ; Podocytes/drug effects*

OBJECTIVETo explore the effects of Astragalus membranaceus (AM) on the cytokines secretion of peripheral dendritic cells (DC), including interleukin-10, -12, and -18 (IL-10, IL-12 and IL-18), in children with Henoch-Schonlein purpura (HSP) in the acute phase; and to study the immunological regulation mechanism of AM.

METHODSPeripheral blood mononuclear cells (PBMC) were obtained from 28 children with acute HSP by density gradient centrifugation, and each sample was divided into two parts, one untreated and one treated with AM. All cells were developed to mature DC through treating with recombinant human granulocyte macrophage colony stimulating factor (GM-CSF), interleukin-4 (IL-4) and tumor necrosis factor-alpha (TNF-alpha). Expression of CD83 in the surface of mature DC was detected by flow cytometry, and levels of IL-10, IL-12 and IL-18 in the supernatant were measured by ELISA.

RESULTSThe supernatant level of IL-12 was higher [(141.58 +/- 100.19) ng/L vs (96.18 +/- 76.65) ng/L, t = 3.90, P<0.01], while levels of IL-10 and IL-18 were lower (t = 2.70, P<0.05; t = 4.07, P<0.01) in AM treated PBMCs than those in the untreated ones.

CONCLUSIONAM can correct the immunologic dysfunction of HSP children through increasing the IL-12, and decreasing the IL-10 and IL-18 secretions of PBMCs.

Adolescent ; Astragalus membranaceus ; Child ; Child, Preschool ; Dendritic Cells ; drug effects ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Female ; Humans ; Interleukin-10 ; blood ; Interleukin-12 ; blood ; Interleukin-18 ; blood ; Leukocytes, Mononuclear ; metabolism ; Male ; Purpura, Schoenlein-Henoch ; blood ; immunology

OBJECTIVETo study the effect of interleukin (IL)-18 ASPODN on regeneration of allogeneic partial liver graft in rats.

METHODSNinety donor SD rats and ninety recipient LEW rats were randomly divided into 3 groups: 50% partial liver transplantation group (PLT group); PLT+IL-18 antisense phosphorothioate oligodeoxynucleotide (ASPODN) treatment group (IL-18 ASPODN group) and PLT+IL-18 SPODN treatment group (IL-18 SPODN group) in which liposomes encapsulated IL-18 ASPODN or IL-18 SPODN were intravenous injection every day after PLT. BrdU labeling of hepatocytes, expression of IL-18 protein and IFN-gamma mRNA in liver graft, and serum level of IFN-gamma were measured with immunohistochemistry analysis, Western blotting, semi-quantification RT-PCR, and ELISA, respectively.

RESULTSAlthough regeneration of liver graft from each group peaked 72 hour after transplantation, BrdU labeling of hepatocytes in IL-18 ASPODN group (58.3%+/-7.5%) were significantly higher than those of PLT group (31.6%+/-6.7%) (t=6.503, P<0.001) and IL-18 SPODN group (33.4%+/-5.5%) (t=6.558, P<0.001). Expression of IL-18 protein and IFN-gamma mRNA in liver graft, and serum level of IFN-gamma in IL-18 ASPODN group from 48 hour, 72 hour and 96 hour after transplantation were significantly suppressed compared with PLT group (IL-18protein: t=2.950, t=5.916, t=7.947, P<0.05, P<0.001; INF-gamma mRNA: t=2.558, t=6.292, t=8.925, P<0.05, P<0.001; IFN-gamma level: t=16.998, t=15.483, t=54.723, P<0.001) and IL-18 SPODN group (IL-18 protein: t=2.845, t=6.062, t=6.973, P<0.05, P<0.001; INF-gamma mRNA: t=3.117, t=6.154, t=8.738, P<0.05, P<0.001; IFN-gamma level: t=14.531, t=18.139, t=46.924, P<0.001).

CONCLUSIONIL-18 ASPODN could promote hepatocyte regeneration of allogeneic partial liver graft by the suppression of IL-18 and IFN-gamma production.

Animals ; Hepatocytes ; physiology ; Interferon-gamma ; biosynthesis ; Interleukin-18 ; antagonists & inhibitors ; genetics ; Liver Regeneration ; Male ; Oligodeoxyribonucleotides, Antisense ; pharmacology ; Rats ; Rats, Inbred Lew ; Rats, Sprague-Dawley ; Transplantation, Homologous

Animals ; Antibodies, Anti-Idiotypic ; therapeutic use ; Disease Models, Animal ; Doxorubicin ; toxicity ; Immunotherapy ; Interleukin-18 ; immunology ; pharmacology ; Male ; Mice ; Nephrosis, Lipoid ; chemically induced ; pathology ; therapy

Animals ; Concanavalin A ; adverse effects ; Immunoglobulin G ; blood ; Interleukin-18 ; blood ; immunology ; pharmacology ; Liver Cirrhosis ; blood ; chemically induced ; prevention & control ; Mice ; Mice, Inbred BALB C

OBJECTIVETo explore the effect of p38MAPK signaling pathway in interleukin-18-induced transdifferentiation in renal proximal tubular cells.

METHODSHuman proximal tubular epithelial cell line (HK-2 cells) was cultured in vitro. After preincubated with SB203580 (0, 5, 10, 20 micromol/L) for 30 minutes, cells were exposed to IL-18 (100 ng/ml) for 24, 48 and 72 hours respectively. The expressions of a-smooth actin (alpha-SMA) in cultured HK-2 cells were assessed by RT-PCR and ELISA.

RESULTSIL-18-induced expressions of a-SMA mRNA and protein were inhibited obviously by a dose-dependent manner when HK-2 cells were incubated with SB203580 (0, 5, 10, 20 micromol/L) and IL-18 (100 ng/ml) for different time (P < 0.05).

CONCLUSIONIL-18-induced transdifferentiation of renal tubular epithelial cells (RTECs) is suppressed obviously by blocking p38MAPK signaling pathways. IL-18-induced transdifferentiation of RTECs is probably mediated, at least in part, through the activation of p38MAPK signaling pathways.

Cell Line ; Cell Transdifferentiation ; drug effects ; Enzyme Inhibitors ; pharmacology ; Epithelial Cells ; cytology ; Fibrosis ; physiopathology ; Humans ; Imidazoles ; pharmacology ; Interleukin-18 ; pharmacology ; Kidney ; pathology ; Kidney Tubules, Proximal ; cytology ; MAP Kinase Signaling System ; Myofibroblasts ; cytology ; Pyridines ; pharmacology ; p38 Mitogen-Activated Protein Kinases ; metabolism