OBJECTIVETo study the effects and immunoregulation mechanism of the traditional Mongolian medicine Wuweifengshi capsule on adjuvant arthritis (AA).

METHODWister rats were divided into several groups: normal group, AA model group, Wuweifengshi capsule groups (with low, moderate, high dose of 0.2, 0.4, 0.8 g x kg(-1) x d(-1) respectively), and Zhonglun-5 group (original dose of 1.68 g x kg(-1) x d(-1)). The edema degree, the level of IL-1beta, TNF-alpha, PGE2, NO and MDA and the activity of SOD in serum were detected. Through cell culture, the effects of the medicine on AA rat's splenic cell's multiplication capacity were studied. The influence of celiac macrophage cell culture fluid of AA rats' on C57BL/6J mice thymic cell multiplication capacity under the medicine was evaluated.

RESULTWuweifengshi capsule showed an inhibiting function on the level of IL-1beta, TNF-alpha, PGE2, NO and increased the activity of SOD in serum, but showed no significant influence on MDA. It also inhibited the AA rat's splenic cell's multiplication capacity and the influence of celiac macrophage cell culture fluid of AA rat's on C57BL/6J mice thymic cell multiplication capacity.

CONCLUSIONThe anti-AA effect of Wuweifengshi capsule is possibly due to its inhibition of relevant cytokines and its adjustment of corresponding enzyme's activity and immunization organ's cell multiplication capacity.

Animals ; Arthritis, Experimental ; drug therapy ; immunology ; metabolism ; pathology ; Capsules ; Dehydroascorbic Acid ; analogs & derivatives ; blood ; Dinoprostone ; metabolism ; Disease Models, Animal ; Edema ; drug therapy ; Female ; Interleukin-1beta ; metabolism ; Lymphocytes ; immunology ; metabolism ; Macrophages, Peritoneal ; metabolism ; Male ; Medicine, Mongolian Traditional ; Mice ; Nitric Oxide ; metabolism ; Rats ; Spleen ; cytology ; metabolism ; Superoxide Dismutase ; blood ; Tumor Necrosis Factor-alpha ; metabolism

Inflammasome is a large protein complex activated upon cellular stress or microbial infection, which triggers maturation of pro-inflammatory cytokines interleukin-1β and interleukin-18 through caspase-1 activation. Nod-like receptor family protein 3 (NLRP3) is the most characterized inflammasome activated by various stimuli. However, the mechanism of its activation is unclear and its exact cellular localization is still unknown. We examined the potential co-localization of NLRP3 inflammasome with mitochondria and seven other organelles under adenosine triphosphate, nigericin or monosodium urate stimulation in mouse peritoneal macrophages using confocal microscopy approach. Our results revealed that the activated endogenous apoptosis-associated speck-like protein containing a CARD (ASC) pyroptosome forms in the cytoplasm and co-localizes with NLRP3 and caspase-1, but not with any of the organelles screened. This study indicates that the ASC pyroptosome universally localizes within the cytoplasm rather than with any specific organelles.
Adenosine Triphosphate ; pharmacology ; Animals ; Apoptosis Regulatory Proteins ; CARD Signaling Adaptor Proteins ; Carrier Proteins ; analysis ; metabolism ; Caspase 1 ; analysis ; metabolism ; Cytoplasm ; metabolism ; Cytoskeletal Proteins ; analysis ; metabolism ; Inflammasomes ; analysis ; metabolism ; Macrophages, Peritoneal ; cytology ; drug effects ; metabolism ; Mice ; Mice, Inbred C57BL ; Microscopy, Confocal ; Mitochondria ; metabolism ; NLR Family, Pyrin Domain-Containing 3 Protein ; Nigericin ; pharmacology ; Uric Acid ; pharmacology

Pannexin-1 (Panx1) forms nonselective large channel in cell plasma membrane and has been shown to be associated with NLRP3 inflammasome activation, ATP release and phagocytes recruitment. In the current study, by manipulation of Panx1 expression in human myeloid cells and application of Panx1 deficient mice, we failed to find a correlation between Panx1 and NLRP3 inflammasome activation, although an interaction between these two proteins was evident. However, in thioglycollate induced peritonitis, Panx1 deficient mice showed much more phagocytes infiltration. Further analyses showed that mice deficient for Panx1 exhibited enlarged F4/80(low)Gr1(-)Ly6C(-)cell population in the peritonea. Our study thus reveals an important role for Panx1 in regulation of peritoneal cell population and peritonitis development.
Animals ; Carrier Proteins ; metabolism ; Cell Line ; Connexins ; antagonists & inhibitors ; deficiency ; genetics ; metabolism ; HEK293 Cells ; Humans ; Inflammasomes ; metabolism ; Macrophages ; cytology ; metabolism ; Mice ; Mice, Inbred C57BL ; NLR Family, Pyrin Domain-Containing 3 Protein ; Nerve Tissue Proteins ; antagonists & inhibitors ; deficiency ; genetics ; metabolism ; Peritoneal Cavity ; cytology ; Peritonitis ; chemically induced ; metabolism ; pathology ; RNA Interference ; RNA, Small Interfering ; metabolism ; Thioglycolates ; toxicity

ATP-binding cassette transporter A1 (ABCA1) promotes cholesterol and phospholipid efflux from cells to lipid-poor apolipoprotein A-I (apoA-I), and plays a key role in the initial steps of the whole process of reverse cholesterol transport (RCT). Upregulation of ABCA1 is beneficial for atherosclerosis (AS) prevention and/or therapy, which indicated that ABCA1 was a target for anti-AS drug development. In the previous study, a high-throughput screening method was established using ABCA1p-LUC HepG2 cell line to find the upregulators of ABCA1. In the present study, compound 2030421B was found using this method, with EC50 of 0.50 microg x mL(-1). The compound was further identified as an upregulator of ABCA1 expression by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting analysis. Studies also showed that the 2030421B could induce apoA-I-mediated cholesterol efflux and inhibit lipids uptake into mouse peritoneal macrophages RAW264.7.
ATP Binding Cassette Transporter 1 ; ATP-Binding Cassette Transporters ; genetics ; metabolism ; Animals ; Anticholesteremic Agents ; administration & dosage ; chemistry ; pharmacology ; Apolipoprotein A-I ; metabolism ; Benzaldehydes ; administration & dosage ; chemistry ; pharmacology ; Biological Transport ; Cells, Cultured ; Cholesterol ; secretion ; Dose-Response Relationship, Drug ; Hep G2 Cells ; High-Throughput Screening Assays ; Humans ; Lipid Metabolism ; Lipids ; analysis ; Macrophages, Peritoneal ; cytology ; metabolism ; Mice ; Molecular Structure ; RNA, Messenger ; Up-Regulation ; drug effects

OBJECTIVETo observe the effect of andrographolide on the activation of mitogen-activated protein kinases (MAPKs) and expression of nuclear factor-κB (NF-κB) in macrophage foam cells.

METHODSThe mouse peritoneal macrophages were cultured in the media in the presence of oxidized low-density lipoprotein (ox-LDL), ox-LDL+andrographolide, or neither (control). The phosphorylation of MAPK molecules (p38MAPK, JNK, ERK1/2) and the expressions of NK-κB p65 were examined by Western blot.

RESULTSAs compared with cells in the control group, the expressions of phospho-p38 and NF-κB p65 were increased in the cells cultured with either ox-LDL or ox-LDL+andrographolide (P<0.01), but attenuated significantly in the presence of ox-LDL+ andrographolide when compared with ox-LDL (P<0.05). The phospho-JNK increased in the presence of either ox-LDL or ox-LDL+andrographolide when compared with control cells (P<0.01), but no significant difference existed between ox-LDL and ox-LDL+andrographolide (P>0.05). The expression of phospho-ERK1/2 was increased in the presence of ox-LDL compared with the control cells (P<0.01), but no significant differences existed between the cells cultured in the presence of ox-LDL+andrographolide and the control medium (P>0.05).

CONCLUSIONSAndrographolide could inhibit the activation of ERK1/2, p38MAPK and NK-κB induced by ox-LDL in macrophage foam cells, which might be one of its mechanisms in preventing atherosclerosis.

Animals ; Anti-Inflammatory Agents ; pharmacology ; Atherosclerosis ; immunology ; metabolism ; prevention & control ; Cells, Cultured ; Diterpenes ; pharmacology ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Foam Cells ; cytology ; drug effects ; enzymology ; JNK Mitogen-Activated Protein Kinases ; metabolism ; Lipoproteins, LDL ; metabolism ; MAP Kinase Signaling System ; drug effects ; immunology ; Macrophages, Peritoneal ; cytology ; drug effects ; enzymology ; Mice ; Mice, Inbred Strains ; NF-kappa B ; metabolism ; Vasculitis ; drug therapy ; immunology ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism

In this work, we used a preparation of diminazene, which belongs to the group of aromatic diamidines. This compound acts on the causative agents of blood protozoan diseases produced by both flagellated protozoa (Trypanosoma) and members of the class Piroplasmida (Babesia, Theileria, and Cytauxzoon) in various domestic and wild animals, and it is widely used in veterinary medicine. We examined the behavior of water-disperse diminazene (immobilized in Tween 80 micelles) at the cellular and organismal levels. We assessed the interaction of an aqueous and a water-disperse preparation with cells of the reticuloendothelial system. We compared the kinetic parameters of aqueous and water-disperse diminazene in sheep erythrocytes and plasma. The therapeutic properties of these two preparations were also compared. We found that the surface-active substances improved intracellular penetration of the active substance through interaction with the cell membrane. In sheep blood erythrocytes, micellar diminazene accumulated more than its aqueous analog. This form was also more effective therapeutically than the aqueous analog. Our findings demonstrate that use of micellar diminazene allows the injection dose to be reduced by 30%.
Animals ; Babesiosis/drug therapy/veterinary ; Diminazene/metabolism/*pharmacokinetics ; Dose-Response Relationship, Drug ; Female ; Macrophages, Peritoneal/cytology/metabolism ; Male ; Micelles ; Polysorbates ; Rats ; Sheep/*blood ; Sheep Diseases/drug therapy ; Trypanocidal Agents/*pharmacokinetics

OBJECTIVETo study effects of polysaccharide of Radix Ranunculi Ternati (PRT) on immunological function and anti-oxidation activity of mouse.

METHODCell proliferations of splenocyte, thymocyte and peritoneal macrophage were measured by MTT colorimetry. The phagocytic function of peritoneal macrophage was measured by neutral red colorimetric method. The disoxidation power of PRT was measured by Prussian blue method. The clearing effect of PRT on hydroxyl radical was measured by salicylic acid capture method. The clearing effect of PRT on superoxide anion free radical was measured by pyrogallol auto oxidation method.

RESULTPRT among 25-400 mg x L(-1) could enhance thymocytes and spleen lymphocyte proliferation and macrophage phagocytosis. PRT(200 mg x L(-1)) has the strongest macrophage proliferation. PRT in different concentration has shown some disoxidation effects. PRT in 8 g x L(-1) has nearly the same ability of clearing x OH by Vit C with the same concentration. The clearance rate of PRT on O2*- is 95.39%.

CONCLUSIONPRT can enhance the cell proliferation capability of thymocytes, spleen lymphocytes and peritoneal macrophages. PRT can enhance macrophage phagocytosis in a dose-response relationship. PRT has saome disoxidation power and strong ability of clearing x OH and O2*-.

Animals ; Antioxidants ; analysis ; pharmacology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Macrophages, Peritoneal ; cytology ; drug effects ; immunology ; Male ; Mice ; Oxidation-Reduction ; drug effects ; Phagocytosis ; drug effects ; Polysaccharides ; immunology ; pharmacology ; Ranunculus ; chemistry ; Spleen ; cytology ; drug effects ; immunology ; Thymus Gland ; cytology ; drug effects ; immunology

OBJECTIVETo study the inhibitory effect of dexamethasone (DEX) on myeloid differentiation factor 88 (MyD88) and tumor necrosis factor-alpha (TNF-alpha) expression in mouse peritoneal macrophages in innate immune response to Penicillium marneffei (PM).

METHODSMouse peritoneal macrophages were cultured in the presence of heat-inactivated yeast-phase PM with or without DEX, and the protein and mRNA expressions of MyD88 in the macrophages were detected using Western blotting and real-time PCR, respectively. TNF-alpha in the cell culture supernatant was measured with enzyme-linked immunosorbent assay.

RESULTSDEX suppressed TNF-alpha production by the macrophages co-cultured with PM. The expressions of MyD88 were up-regulated by PM stimulation, whose effect was inhibited by the application of DEX.

CONCLUSIONThe inhibitory effect of DEX on PM-induced proinflammatory responses of the macrophage is directly associated with the inhibition of MyD88 expression.

Animals ; Cells, Cultured ; Dexamethasone ; pharmacology ; Macrophages, Peritoneal ; cytology ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Myeloid Differentiation Factor 88 ; drug effects ; genetics ; metabolism ; Penicillium ; growth & development ; Tumor Necrosis Factor-alpha ; drug effects ; genetics ; metabolism

OBJECTIVETo study the anti-inflammatory mechanisms of total glycosides of Acanthopanax Giraldii (TGA).

METHODSThe changes of prostaglandin E(2)(PGE(2)), tumor necrosis factor (TNF-alpha), nitric oxide (NO), and expressions of COX-1 mRNA and COX-2 mRNA in BALB/c mouse macrophages were observed by the radioimmunoassay, ELISA and nitric acid reduction and RT-PCR in the presence or absence of TGA.

RESULTS(1) TGA could significantly decrease the production of PGE(2)and NO in mouse peritoneal macrophages. The inhibitory rate to LPS-induced PGE(2)production was 87% (TGA 100 mg/L, P<0.05, vs. LPS) and 62% (TGA 20 mg/L, P<0.05, vs. LPS), respectively. The inhibitory rate of NO production in mouse peritoneal macrophages was 49% (TGA 100 mg/L, P<0.05, vs. LPS) and 21% (TGA 20 mg/L, P<0.05 vs. LPS), respectively. TGA could not inhibit LPS-induced TNF-alpha production in mouse peritoneal macrophages. (2) TGA also inhibited the expression of COX-1 and COX-2 mRNA in RAW264.7 cells. The inhibitory rate of TGA to COX-1 mRNA was 22% (TGA 100 mg/L, P<0.05, vs. blank). The inhibitory rate of TGA to COX-2 mRNA was 55% (TGA 20 mg/L, P<0.05, vs. LPS) and 100% (TGA 100 mg/L, P<0.01 vs. LPS), respectively.

CONCLUSIONThe anti-inflammatory mechanisms of TGA for inhibiting the production of NO and PGE(2)are through inhibiting COX-2 mRNA expression without TNF-alpha changes.

Animals ; Anti-Inflammatory Agents ; pharmacology ; Cell Line ; Cyclooxygenase 1 ; genetics ; Cyclooxygenase 2 ; genetics ; Dinoprostone ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Eleutherococcus ; Female ; Gene Expression Regulation, Enzymologic ; drug effects ; Glycosides ; pharmacology ; Lipopolysaccharides ; pharmacology ; Macrophages, Peritoneal ; cytology ; drug effects ; metabolism ; Mice ; Mice, Inbred BALB C ; Nitric Oxide ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo study the effect of total glucosides of paeony (TGP) on lipopolysaccharides (LPS)-induced nuclear factor-kappaB (NF-kappaB) activation in macrophages.

METHODRat peritoneal macrophages were pre-treated with TGP for 2 h and stimulated with LPS for 20 min or 0.5 h. Inhibitory kappaBalpha (IkappaBalpha) protein in the cytoplasm and NF-kappaB p65 protein in the nuclear were analyzed by western blot. Further, DNA binding activity of NF-kappaB complex was detected.

RESULTTGP enhanced the amounts of IkappaBalpha protein in the cytoplasm and decreased the amounts of NF-kappaB p65 protein in the nuclear of LPS-induced macrophages. TGP also inhibited the LPS-mediated DNA binding activity of NF-kappaB complex in macrophages.

CONCLUSIONTGP can inhibit LPS-induced NF-kappaB activation in macrophages through arresting IKBalpha protein degradation, NF-kappaB p65 protein nuclear translocation and DNA binding activity of NF-kappaB complex.

Animals ; Cell Nucleus ; drug effects ; metabolism ; Cytoplasm ; drug effects ; metabolism ; DNA ; metabolism ; Dose-Response Relationship, Drug ; Glucosides ; pharmacology ; Macrophages, Peritoneal ; cytology ; drug effects ; metabolism ; NF-kappa B ; metabolism ; Paeonia ; chemistry ; Protein Transport ; drug effects ; Rats ; Transcription Factor RelA ; metabolism