OBJECTIVETo study the antalgic and antiphlogistic functions and mechanism of ronggudingtong (RGDT) plaster (traditional Chinese medicine).

METHODSThe painful models were established with hot plate test or acetic acid writhing and the inflammatory models were established with daubing dimethylbenzene on auricle or injecting formaldehyde in toe or synovial envelope to study the antalgic and antiphlogistic functions of RGDT Plaster. The total protein and leukotriene B4(LTB4) in inflammatory exudate were detected to investigate the antalgic and antiphlogistic mechanism of RGDT plaster. The mice were randomly divided into different groups (n = 11), on the basis of drug using, the indexes of pain threshold, swelling degree were observed. Sixty-six mice were used to establish gasbag synovitis model and randomly divided into normal control group,model control group, positive control group (Voltaren gel 0.8 mg/d)and low/medium/high dosage RGDT plaster treating groups(30 mg/d, 60 mg/d, 120 mg/d).

RESULTS30 mg/d, 60 mg/d,120 mg/d RGDT plaster could upgrade the pain thresholds, remit auricular and foot swelling (P < 0.05, P < 0.01), and degrade total protein and LTB4 in inflammatory exudates (P < 0.05, P < 0.01).

CONCLUSIONRGDT plaster has some antalgic and antiphlogistic functions, and one of the mechanisms is depressing synthesis of LTB4.

Analgesics ; pharmacology ; Animals ; Anti-Inflammatory Agents ; pharmacology ; Leukotriene B4 ; metabolism ; Medicine, Chinese Traditional ; Mice ; Pain ; drug therapy

OBJECTIVETo observe the changing contents of leukotriene B4 ( LTB4 ), leukotriene C4 ( LTC4 ), and leukotriene D4 (LTD4 ) of lung tissue in asthmatic rats, and explore the effect of Shuanglong Capsule (SLC) on it.

METHODSSD rats were randomly divided into the nomal group, asthmatic model group, Dexamethasone group and the high, middle and low dose SLC groups. All rats except those in the normal group were sensitized by ovalbumin and challenged with the antigen, and the contents of LTB4, LTC4 and LTD4 in lung tissue of all the groups were measured by reverse phase-high performance liquid chromatography (RP-HPLC) and compared.

RESULTSThe levels of LTB4, LTC4, and LTD4 of asthmatic rats were significantly higher than those of rats in the normal group. Dexamethasone and SLC at the dose of 8. 27 g/kg or 4. 13 g/kg could significantly inhibit the production of leukotrienes of lung tissue in asthmatic rats (P <0.05).

CONCLUSIONSLC can significantly inhibit the formation of inflammatory medium LTs of lung tissue in asthmatic rats, it may be one of the key mechanisms of SLC in anti-asthma and anti-inflammatory action.

Animals ; Anti-Asthmatic Agents ; pharmacology ; Asthma ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Leukotriene B4 ; metabolism ; Leukotriene C4 ; metabolism ; Leukotriene D4 ; metabolism ; Leukotrienes ; metabolism ; Lung ; drug effects ; metabolism ; Rats ; Rats, Sprague-Dawley ; Tablets

OBJECTIVETo study inhibitory the effects of Cryptoporus volvatus ferment substance(CVFS) on leukotriene production in vitro from neutrophils in rats.

METHODSNeutrophil aggregation was induced by intraperitoneal injection of glycogen in rats. After 16 h, intraperitoneal lavage fluid(PLF) was collected and neutrophils were removed. Neutrophils were stimulated by calcium ionophore A23187 in vitro to produce leukotriene B(4), C(4), D(4). The concentrations of leukotriene B(4), C(4) and D(4) were measured by reversed-phase high-performance liquid chromatography(HPLC).

RESULTCVFS at 0.25, 1, 4 mg x L(-1)decreased leukotriene B(4), C(4), D(4) release from neutrophils in a concentration-dependent manner. Inhibitory rate of CVFS 0.25, 1, 4 mg x L(-1 )on A23187-induced leukotriene B(4) production was 27.4%, 54.2% and 78.8%(P<0.05), respectively. Inhibitory rate of leukotriene C(4) production was 65.1%, 74.3 and 79.0%(P<0.05), respectively. Inhibitory rate of leukotriene D(4) production was 55.6%, 60.9% and 72.8%(P<0.05), respectively.

CONCLUSIONThe results suggest that suppression of leukotriene release may be a mechanism of the anti-inflammation and anti-asthma effects of CVFS.

Animals ; Anti-Asthmatic Agents ; pharmacology ; Dose-Response Relationship, Drug ; Female ; Fermentation ; Leukotriene B4 ; secretion ; Leukotriene C4 ; secretion ; Leukotriene D4 ; secretion ; Male ; Neutrophils ; drug effects ; physiology ; Polyporaceae ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo observe the inhibitory effect of mizolastine on substance P(SP)-induced production of leukotriene B(4) (LTB(4)) and interleukin 5 (IL-5) in mouse skin.

METHODSMice were fed with different doses of mizolastine or other control drugs, 30 min after administration animals were injected intradermally with SP on the back. The treated skin samples were taken and competitive enzyme-link immunoassay (ELISA) method was applied to detect LTB (4) and IL-5 in the skin samples.

RESULTThe LTB(4) and IL-5 levels in 10 mg/kg mizolastine group were (1.23 +/-0.29)pg/ml and (34.28 +/-11.00)pg/ml, respectively, which were lower than those in saline control group [(5.52+/-1.88)pg/ml and (179.62 +/-46.25)pg/ml respectively] or loratadine group [(3.89+/-1.27)pg/ml and (127.74 +/-43.27)pg/ml respectively]. No significant difference was found between 10 mg/kg mizolastine group and dexamethasone group (P=0.161 and P=0.508).

CONCLUSIONMizolastine might inhibit the production of LTB(4) and IL-5 induced by substance P in mouse skin, suggesting that anti-inflammatory effect and the blockade of histamine H1 receptors might be involved in its anti-pruritic mechanisms.

Animals ; Benzimidazoles ; pharmacology ; Female ; Histamine H1 Antagonists ; pharmacology ; Interleukin-5 ; biosynthesis ; Leukotriene B4 ; biosynthesis ; Male ; Mice ; Mice, Inbred BALB C ; Skin ; metabolism ; Substance P ; antagonists & inhibitors

Primary ciliary dyskinesia is characterized by chronic upper and lower respiratory infections which are caused by the grossly impaired ciliary transport. Since the cilia and neutrophils both utilize microtubular system for their movement, it has been speculated that neutrophil motility such as chemotaxis might be impaired in patients with primary ciliary dyskinesia. Neutrophils were purified from whole blood from 16 patients with primary ciliary dyskinesia and from 15 healthy controls. Chemotactic responses of neutrophils to leukotriene B4 (LTB4), complement 5a (C5a), and formylmethion-ylleucylphenylalanine (fMLP) were examined using the under agarose method. The chemotactic differentials in response to LTB4, C5a, and fMLP in neutrophils from the patient group were significantly lower than the corresponding values in neutrophils from the control group (p<0.05 for all comparisons). The difference in chemotactic index between the two groups was statistically significant for LTB4 and fMLP (p<0.05 for both comparisons), but not for C5a (p=0.20). Neutrophils from patients with primary ciliary dyskinesia showed a decreased chemotactic response as compared with those from normal subjects. It is concluded that the increased frequency of respiratory tract infection in patients with primary ciliary dyskinesia is possibly due to the defective directional migration of neutrophils, as well as to the defective mucociliary clearance of the airways.
Adolescent ; Chemotactic Factors/pharmacology ; Chemotaxis* ; Child ; Cilia/ultrastructure ; Comparative Study ; Complement 5a/pharmacology ; Dose-Response Relationship, Drug ; Dynein ATPase/chemistry ; Human ; Kartagener Syndrome/blood* ; Kartagener Syndrome/classification ; Leukotriene B4/pharmacology ; Male ; N-Formylmethionine Leucyl-Phenylalanine/pharmacology ; Neutrophils/physiology* ; Neutrophils/ultrastructure

Leukotriene B4(LTB4), derived from arachidonic acid, is a potent chemotactic agent and activating factor for hematopoietic cells. In addition to host defense in vivo, several eicosanoids have been reported to be involved in stem cell differentiation or proliferation. In this study, we investigated the effect of LTB4 on human cord blood CD34+ hematopoietic stem cells (HSCs). LTB4 was shown to induce proliferation of HSC and exert anti-apoptotic effect on the stem cells. Blockade of interaction between LTB4 and its receptor enhanced self-renewal of the stem cells. Effect of LTB4 on differentiation of CD34+ HSCs were confirmed by clonogenic assays, and induction of the expression of BLT2 (the low- affinity LTB4 receptor), during the ex vivo expansion was confirmed by reverse transcription-PCR. Our results suggest that LTB4-BLT2 interaction is involved in the cytokine-induced differentiation and ex vivo expansion of hematopoietic stem cells.
Antigens, CD34/metabolism ; Apoptosis/drug effects ; Cell Differentiation/drug effects ; Cell Proliferation/drug effects ; Fetal Blood/cytology/drug effects ; Hematopoietic Stem Cells/*drug effects/metabolism ; Humans ; Leukotriene B4/*pharmacology ; Receptors, Leukotriene B4/genetics/metabolism ; Research Support, Non-U.S. Gov't ; Reverse Transcriptase Polymerase Chain Reaction ; *Signal Transduction

AIMBLT1 and BLT2 were both recently cloned and identified as two subtypes of leukotrine B4 (LTB4) receptors. With the usage of U-75302 and LY255283, the specific antagonists of BLT1 and BLT2 respectively, the involvement of BLT1 and BLT2 in the inflammatory and immunological responses was in vitro explored.

METHODS(1) To investigate inhibition of U-75302 and LY255283 on the proliferation of rat synovial cells, 3H-TdR incorporation into the cells was quantified. (2) Flow cytometric assay for interferon-gamma (IFN-gamma) and interleukine 4 (IL-4) profiles in CD4+ T lymphocytes from rat spleen was carried out to determine the ratio of Th1/Th2.

RESULTS(1) For inhibition on rat synovial cells proliferation, U-75302 exerted its effect only at a high concentration of 10 micromol/L and LY255283 at the concentrations of 10 micromol/L-10 micromol/L. (2) Both U-75302 and LY255283 could elevate the percentage of Th2, but could not influence that of Th1.

CONCLUSIONBLT1 and BLT2 were involved in the synovial cells proliferation change the ratio of Th1/Th2. Their meaning served as targets for prevention and treatment of infectious diseases should be emphasized.

Animals ; Cell Line ; Cell Proliferation ; drug effects ; Fatty Alcohols ; pharmacology ; Glycols ; pharmacology ; Inflammation ; immunology ; Male ; Rats ; Rats, Wistar ; Receptors, Leukotriene B4 ; antagonists & inhibitors ; physiology ; Synovial Membrane ; cytology ; immunology ; Tetrazoles ; pharmacology ; Th1-Th2 Balance

OBJECTIVETo study action of Cryptoporus volvatus ferment substance (CVFS) on leukotriene production of polymorphonuclear leukocytes in rats.

METHODSThe level of slow reaction substance (SRS) and leukotriene B4 (LTB4) in polymorphonuclear leukocytes (PMNs) in rats in vitro were determined with bioassay and HPLC.

RESULTSCVFS 0.9, 2.7 g.kg-1 by ig significantly inhibited SRS and LTB4 production in PMNs in rats in vivo.

CONCLUSIONThe inhibition effect of CVFS on SRS and LTB4 release may be related to its mechanism of anti-inflammation and anti-asthma.

Animals ; Anti-Asthmatic Agents ; pharmacology ; Anti-Inflammatory Agents, Non-Steroidal ; pharmacology ; Cell Separation ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Guinea Pigs ; Leukotriene B4 ; metabolism ; Male ; Neutrophils ; metabolism ; Polyporaceae ; chemistry ; Rats ; Rats, Sprague-Dawley ; SRS-A ; metabolism

OBJECTIVETo explore the regularity of recipe composition by observing inhibitory effects of disassembled compositions of Mahuang decoction (MHD) on chemotaxis and leukotriene production from neutrophils in rats.

METHODNeutrophil aggregation was induced by intraperitoneal injection of glycogen in rats. Intraperitoneal lavage fluid (PLF) was collected and neu-trophils were removed. Neutrophils were stimulated by calciumionophore A23187 in vitro to produce leukotriene B4. The concentrations of leukotriene B4 was measured by reversed-phase high performance liquid chromatography(HPLC), chemotatic chamber assay was used to investigate the regulative role of MHD on chenmotaxis of the neutrophils in response to LPS stimulation.

RESULTDisassembled compositions of MHD could inhibite chemotaxis and leukotriene production from neutrophils in rats. Inhibitory effects of MHD on mast cells were different.

CONCLUSIONMHD has significantly inhibitory effects on chemotaxis and leukotriene production from neutrophils in rats. The original formula (MHD) works best. These results have confirmed the rationality and scientific level of MHD.

Animals ; Chemotaxis, Leukocyte ; drug effects ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Female ; Leukotriene B4 ; biosynthesis ; Male ; Neutrophils ; drug effects ; secretion ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley

The incidence rates of urinary bladder cancer continue to rise yearly, and thus new therapeutic approaches and early diagnostic markers for bladder cancer are urgently needed. Thus, identifying the key mediators and molecular mechanisms responsible for the survival of bladder cancer has valuable implications for the development of therapy. In this study, the role of BLT2, a receptor for leukotriene B4 (LTB4) and 12(S)-hydroxyeicosatetraenoic acid (HETE), in the survival of bladder cancer 253J-BV cells was investigated. We found that the expression of BLT2 is highly elevated in bladder cancer cells. Also, we observed that blockade of BLT2 with an antagonist or BLT2 siRNA resulted in cell cycle arrest and apoptotic cell death, suggesting a role of BLT2 in the survival of human bladder cancer 253J-BV cells. Further experiments aimed at elucidating the mechanism by which BLT2 mediates survival revealed that enhanced level of reactive oxygen species (ROS) are generated via a BLT2-dependent up-regulation of NADPH oxidase members NOX1 and NOX4. Additionally, we observed that inhibition of ROS generation by either NOX1/4 siRNAs or treatment with an ROS-scavenging agent results in apoptotic cell death in 253J-BV bladder cancer cells. These results demonstrated that a 'BLT2-NOX1/4-ROS' cascade plays a role in the survival of this aggressive bladder cancer cells, thus pointing to BLT2 as a potential target for anti-bladder cancer therapy.
*Apoptosis ; Blotting, Western ; Cell Proliferation ; Cells, Cultured ; Gene Expression Regulation, Neoplastic/*physiology ; Humans ; Leukotriene Antagonists/pharmacology ; NADPH Oxidase/antagonists & inhibitors/genetics/metabolism ; Phosphorylation ; RNA, Messenger/genetics ; RNA, Small Interfering/genetics ; Reactive Oxygen Species/*metabolism ; Receptors, Leukotriene B4/antagonists & inhibitors/*genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Signal Transduction ; Tetrazoles/pharmacology ; Up-Regulation ; Urinary Bladder Neoplasms/*genetics/mortality