1.Expression and role of Tc17 cells in mice with neutrophilic asthma.
Min ZHANG ; Guang-Min NONG ; Min JIANG ; Wen-Jie ZHAN
Chinese Journal of Contemporary Pediatrics 2016;18(2):172-176
OBJECTIVETo determine the proportion of Tc17 cells in the lungs of mice with neutrophilic(NEU) asthma, and to investigate the role of Tc17 cells in the pathogenesis of NEU asthma.
METHODSThirty-two C57/B6 mice of clean grade were randomly divided into two groups: NEU asthma and control. The mice in the NEU asthma group were sensitized by airway instillation of ovalbumin (OVA) and lipopolysaccharide (LPS), and challenged with an aerosol of OVA, while those in the control group were sensitized and challenged with normal saline. At 24 hours after the final challenge, bronchoalveolar lavage fluid (BALF) was collected, and the total number and differential counts of nucleated cells and percentage of each type were determined. The lung tissues were separated and hematoxylin-eosin staining was performed to observe the pathological changes of lungs; flow cytometry was applied to determine the percentages of Tc17 and Th17 cells in the lung tissues. Enzyme-linked immunosorbent assay was applied to determine the levels of interleukin-6 (IL-6), transforming growth factor β (TGF-β), and interleukin-17 (IL-17) in BALF.
RESULTSThe NEU asthma group had a significantly higher total number of nucleated cells, a significantly higher percentage of eosinophils, and a significantly higher percentage of neutrophils in BALF than the control group (P<0.01). The NEU asthma group also had significantly higher percentages of Tc17 and Th17 cells than the control group (P<0.01). In the NEU asthma group, the percentage of Tc17 cells was positively correlated with that of Th17 cells (P<0.05). The NEU asthma group had significantly higher concentrations of IL-6, TGF-β, and IL-17 in BALF than the control group (P<0.05).
CONCLUSIONSThe expression of Tc17 cells in the lung tissues increases in mice with NEU asthma, and the increased number of Tc17 cells may be involved in the pathogenesis of NEU asthma. Tc17 cells may play an important role in NEU asthma through IL-17.
Animals ; Asthma ; genetics ; immunology ; CD8-Positive T-Lymphocytes ; immunology ; Female ; Humans ; Interferon-gamma ; immunology ; Interleukin-17 ; genetics ; immunology ; Interleukin-4 ; genetics ; immunology ; Interleukin-6 ; genetics ; immunology ; Lymphocyte Count ; Mice ; Th17 Cells ; immunology
2.Mechanism of tonifying Qi traditional Chinese medicines contained in Yiqi Qingwen Jiedu mixture against influenza immune inflammatory injury.
Hong-Ri XU ; Cheng-Xiang WANG ; Lan WANG ; Ping-An ZHOU ; Ren-Yi YIN ; Liang-Duo JIANG ; Hui-Fang WANG
China Journal of Chinese Materia Medica 2014;39(20):4020-4026
OBJECTIVETo observe the impact of tonifying Qi traditional Chinese medicines contained in Yiqi Qingwen Jiedu mixture on mRNA expression of lung inflammatory cytokines and pulmonary pathological injury of mice infected by influenza virus, in order to discuss the mechanism of tonifying Qi traditional Chinese medicines against pulmonary immune inflammatory injury of infected mice.
METHODIn different time phases after mice were infected with influenza virus FM1, the RT-PCR method was adopted to observe the impact of tonifying Qi traditional Chinese medicines contained in Yiqi Qingwen Jiedu mixture on five inflammatory cytokines TNF-α, IL-1, IL-6, IL-10 and IFN-γ, and the changes in pulmonary pathological injury of mice with viral pneumonia after intervention with tonifying qi traditional Chinese medicines.
RESULT(1) Tonifying Qi traditional Chinese medicines significantly reduced the mRNA expression of TNF-α at 1-5 d and IL-1 mRNA expression at 7 d, may increase IL-1 mRNA expression in mouse lung at 3 d, significantly reduced IL-6 mRNA expression in mouse lung and increased IL-10 mRNA expression at 3-7 d, and significantly increased IFN-γ mRNA expression at 1 d. (2) Tonifying Qi traditional Chinese medicines could significantly inhibited and repaired pulmonary immune inflammatory injury of mice infected by FM1, which was most remarkable at 3-7 d after the infection with influenza virus FM1.
CONCLUSIONTonifying Qi traditional Chinese medicines contained in Yiqi Qingwen Jiedu mixture could resist pulmonary immune inflammatory injury and repair inflammatory injury by regulating the mRNA expression of imbalance inflammatory cytokines of organisms infected with influenza virus.
Animals ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Influenza A virus ; drug effects ; immunology ; Influenza, Human ; drug therapy ; genetics ; immunology ; Interferon-gamma ; genetics ; immunology ; Interleukin-1 ; genetics ; immunology ; Interleukin-10 ; genetics ; immunology ; Interleukin-6 ; genetics ; immunology ; Lung ; immunology ; virology ; Male ; Mice ; Mice, Inbred BALB C ; Tumor Necrosis Factor-alpha ; genetics ; immunology
3.Cytokine Production by Whole Blood Cells: Relationship to Interleukin Gene Polymorphism and Bone Mass.
Jung Gu KIM ; Seung Yup KU ; Kyung Sil LIM ; Byung Chul JEE ; Chang Suk SUH ; Seok Hyun KIM ; Young Min CHOI ; Shin Yong MOON
Journal of Korean Medical Science 2005;20(6):1017-1022
The aims of this study were to investigate the relationships between the production of interleukin-1 (IL-1), and IL-6 system by whole blood cells, and bone mineral density (BMD), and polymorphisms in IL-1 system and IL-6 gene in postmenopausal Korean women. The production of IL-1alpha, IL-1beta, IL-1 receptor antagonist (IL-1ra), IL-6, and soluble IL-6 receptor (sIL-6r) by lipopolysaccharide-stimulated whole blood cells was measured by ELISA in 110 subjects. Serum osteocalcin, C-telopeptide of type I collagen, and BMD at lumbar spine and proximal femur were measured. IL-1alphaC(-889)T polymorphism, IL-1beta C(-511)T polymorphism, 86-base pair variable number tandem repeat polymorphism in the IL-1ra gene, and IL-6 C(-634)G polymorphism were analyzed. The production of IL-1beta correlated positively with BMD at femoral neck, whereas the production of other ILs did not correlate with BMD at the skeletal sites examined. No significant differences in the production of ILs were observed among normal, osteopenic and osteoporotic postmenopausal women, and among the different IL system polymorphisms groups studied. No correlation between bone turnover markers and the production of ILs was noted. In conclusion IL-1beta may regulate bone metabolism at femoral neck, and the IL system polymorphism do not affect the production of ILs by whole blood cells.
Aged
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Blood Cells/drug effects/immunology
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Bone Density/*genetics/*immunology
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Bone Diseases, Metabolic/blood/genetics/immunology
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Cytokines/*biosynthesis/blood
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Female
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Humans
;
In Vitro
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Interleukin-1/biosynthesis/blood/genetics
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Interleukin-6/biosynthesis/blood/genetics
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Interleukins/*genetics
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Lipopolysaccharides/pharmacology
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Middle Aged
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Osteoporosis, Postmenopausal/blood/genetics/immunology
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*Polymorphism, Genetic
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Receptors, Interleukin-6/biosynthesis/blood/genetics
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Research Support, Non-U.S. Gov't
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Sialoglycoproteins/biosynthesis/blood/genetics
4.Study on in vitro anti-inflammatory activity of total flavonoids from Glycyrrhizae Radix et Rhizoma and its ingredients.
Xiao-Lu YANG ; Duo LIU ; Ka BIAN ; Dan-Dan ZHANG
China Journal of Chinese Materia Medica 2013;38(1):99-104
OBJECTIVETo examine the anti-inflammatory mechanism of total flavonoids of Glycyrrhizae Radix et Rhizoma (TFGR) and its ingredient on IFN-gamma and LPS-induced macrophage RAW264.7.
METHODSolvent extraction and macroporous resin enrichment were adopted for preparing ethanol extracts of Glycyrrhizae Radix et Rhizoma, components and total flavonoids. Ultraviolet spectroscopy was used to determine the content of total flavonoids. An IFN-gamma and LPS-induced cell inflammatory model was established. Griess reaction was used for detecting the effect of extracts at all levels and flavonoid monomers on nitrite content in cell culture supernatant. FRAP was used for measuring anti-oxidation capacity. RT-PCR was used for determining the effect of TFGR and isoliquiritigenins on intracellular inducible nitric oxide synthase iNOS, COX-2, IL-6 and PPAR-gamma. Western blot was used for detecting the effect of TFGR and isoliquiritigenins on iNOS, COX-2 and MAPK signal transduction pathways.
RESULTCompared with other extracts, ethyl acetate fractions from Glycyrrhizae Radix et Rhizoma showed the highest inhibition ratio on nitrite content at the same concentration. After being enriched with macroporous resin, TFGR (60. 08% of liquiritin) of ethyl acetate extracts from Glycyrrhizae Radix et Rhizoma showed dose-dependence, and inhibited the nitrite content in cell culture supernatant, which was superior to ethyl acetate extracts, and had the protective effect on post-stimulated cell activity, with a stronger total anti-oxidation than other extracts. TFGR inhibited iNOS, IL-6 mRNA, protein expressions of iNOS, COX-2 and IL-6. Isoliquiritigenin, a flavonoid monomer, could inhibited iNOS, COX-2 gene and protein expression and gene expressions of IL-1beta and IL-6, and upside-regulated gene expression of PPAR-gamma.
CONCLUSIONActivity-oriented extraction suggests that ethyl acetate fractions from Glycyrrhizae Radix et Rhizoma is one of components with anti-inflammatory activity. TFGR obtained by enriching the active component showed dose-dependence, and inhibited the nitrite content in cell culture supernatant. The anti-inflammatory effect is partially achieved by regulating ERK signal pathway and inhibiting iNOS and COX-2 gene and protein expressions through extracellular signals of mitogen activated protein kinases (MAPKs). Specifically, isoliquiritigenin may be a component with TFGR anti-inflammatory activity.
Animals ; Anti-Inflammatory Agents ; pharmacology ; Cell Line ; Drugs, Chinese Herbal ; pharmacology ; Flavonoids ; pharmacology ; Glycyrrhiza ; chemistry ; Interleukin-1beta ; genetics ; immunology ; Interleukin-6 ; genetics ; immunology ; Macrophages ; drug effects ; immunology ; Mice ; NF-kappa B ; genetics ; immunology ; Rhizome ; chemistry
5.Ginsenoside Rk1 suppresses pro-inflammatory responses in lipopolysaccharide-stimulated RAW264.7 cells by inhibiting the Jak2/Stat3 pathway.
Qian YU ; Ke-Wu ZENG ; Xiao-Li MA ; Yong JIANG ; Peng-Fei TU ; Xue-Mei WANG
Chinese Journal of Natural Medicines (English Ed.) 2017;15(10):751-757
The saponin ginsenoside Rk1 is a major compound isolated from ginseng. Ginsenoside Rk1 has been reported to have anti-inflammatory and anti-tumor properties and to be involved in the regulation of metabolism. However, the effect and mechanism of anti-inflammatory action of ginsenoside Rk1 has not been fully clarified. We investigated whether ginsenoside Rk1 could suppress the inflammatory response in lipopolysaccharide-stimulated RAW264.7 macrophages and to explore its mechanism of the action. RAW264.7 cells were treated with LPS (1 μg·mL) in the absence or the presence of Ginsenoside Rk1 (10, 20, and 40 μmol·L). Then the inflammatory factors were tested with Griess reagents, ELISA, and RT-PCR. The proteins were analyzed by Western blotting. Ginsenoside Rk1 inhibited lipopolysaccharide-induced expression of nitric oxide (NO), interleukin (IL)-6, IL-1β, tumor necrosis factor (TNF)-α, and monocyte chemotactic protein (MCP)-1. Ginsenoside Rk1 inhibited the lipopolysaccharide-stimulated phosphorylation of NF-κB and janus kinase (Jak)2 and signal transducer and activator of transcription (Stat)3 at Ser727 and Tyr705. These data suggested that ginsenoside Rk1 could inhibit expression of inflammatory mediators and suppress inflammation further by blocking activation of NF-κB and the Jak2/Stat3 pathway in LPS-stimulated RAW264.7 cells.
Animals
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Anti-Inflammatory Agents
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pharmacology
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Ginsenosides
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pharmacology
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Interleukin-6
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genetics
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immunology
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Janus Kinase 2
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genetics
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immunology
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Lipopolysaccharides
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pharmacology
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Macrophages
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drug effects
;
immunology
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Mice
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RAW 264.7 Cells
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STAT3 Transcription Factor
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genetics
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immunology
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Tumor Necrosis Factor-alpha
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genetics
;
immunology
6.Anti-hyperuricemic and anti-inflammatory actions of vaticaffinol isolated from Dipterocarpus alatus in hyperuricemic mice.
Yu-Sheng CHEN ; Chao-Jun CHEN ; Wei YAN ; Hui-Ming GE ; Ling-Dong KONG
Chinese Journal of Natural Medicines (English Ed.) 2017;15(5):330-340
The present study was designed to examine the anti-hyperuricemic and anti-inflammatory effects and possible mechanisms of vaticaffinol, a resveratrol tetramer isolated from ethanol extracts of Dipterocarpus alatus, in oxonate-induced hyperuricemic mice. At 1 h after 250 mg·kg potassium oxonate was given, vaticaffinol at 20, 40, and 60 mg·kg was intragastrically administered to hyperuricemic mice once daily for seven consecutive days. Vaticaffinol significantly decreased serum uric acid levels and improved kidney function in hyperuricemic mice. It inhibited hepatic activity of xanthine dehydrogenase (XDH) and xanthine oxidase (XOD), regulated renal mRNA and protein levels of urate transporter 1 (URAT1), glucose transporter 9 (GLUT9), organic anion transporter 1 (OAT1), organic cation transporter 1 (OCT1), OCT2, organic cation/carnitine transporter 1 (OCTN1), and OCTN2 in hyperuricemic mice. Moreover, vaticaffinol markedly down-regulated renal protein levels of NOD-like receptor 3 (NLRP3), apoptosis-associated speck-like (ASC), and Caspase-1, resulting in the reduction of interleukin (IL)-1β, IL-18, IL-6 and tumor necrosis factor-α (TNF-α) levels in this animal model. Additionally, HPLC and LC-MS analyses clearly testified the presence of vaticaffinol in the crude extract. These results suggest that vaticaffinol may be useful for the prevention and treatment of hyperuricemia with kidney inflammation.
Animals
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Anti-Inflammatory Agents
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administration & dosage
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Dipterocarpaceae
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chemistry
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Humans
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Hyperuricemia
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blood
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drug therapy
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immunology
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Interleukin-18
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genetics
;
immunology
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Interleukin-1beta
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genetics
;
immunology
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Interleukin-6
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genetics
;
immunology
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Kidney
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drug effects
;
immunology
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Male
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Mice
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Organic Anion Transport Protein 1
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genetics
;
immunology
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Plant Extracts
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administration & dosage
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Stilbenes
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administration & dosage
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Tumor Necrosis Factor-alpha
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genetics
;
immunology
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Uric Acid
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blood
7.A polysaccharide purified from Radix Adenophorae promotes cell activation and pro-inflammatory cytokine production in murine RAW264.7 macrophages.
Jing-Wen LI ; Yang LIU ; Bao-Hui LI ; Yue-Yang WANG ; Hui WANG ; Chang-Lin ZHOU
Chinese Journal of Natural Medicines (English Ed.) 2016;14(5):370-376
Radix Adenophorae, a traditional Chinese medicine, has been reported to have a variety of biological functions. In the present study, a polysaccharide component, Radix Adenophorae Polysaccharide (RAPS), was purified from Radix Adenophorae by decoloring with ADS-7 macroporous adsorption resin, DEAE-52 cellulose ion-exchange chromatography, and Sephacryl S-300HR gel chromatography, with the purity of 98.3% and a molecular weight of 1.8 × 10(4) Da. The cell viability assay and microscopic examination revealed that RAPS promoted the proliferation and activation of macrophages. At 400 μg·mL(-1), RAPS stimulated RAW264.7 cell proliferation by 1.91-fold compared with the control. Meanwhile, RAPS significantly increased the secretion of pro-inflammatory cytokines (TNF-α and IL-6) in a dose-dependent manner in the supernatant of RAW264.7 cell culture as determined by ELISA. At 400 μg·mL(-1), the production of TNF-iα was 20.8-fold higher than that of the control. Simultaneously, the production of nitric oxide (NO) and the expression of inducible nitric oxide synthase (iNOS) were increased in RAW264.7 cells incubated with RAPS, as measured by Griess assay and Western blot analysis. The NO production of cells treated with RAPS (400 μg·mL(-1)) reached 15.8 μmol·L(-1), which was 30.4-fold higher than that of the control (0.53 μmol·L(-1)). These data suggested that RAPS may enhance the immune function and protect against exogenous pathogens by activating macrophages.
Animals
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Campanulaceae
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chemistry
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Cytokines
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genetics
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immunology
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Immunologic Factors
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pharmacology
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Interleukin-6
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genetics
;
immunology
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Macrophage Activation
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drug effects
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Macrophages
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drug effects
;
immunology
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Mice
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Nitric Oxide
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immunology
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Plant Extracts
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pharmacology
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Polysaccharides
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pharmacology
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Tumor Necrosis Factor-alpha
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genetics
;
immunology
8.Study on biomarker of Tripterygium wilfordii in treatment of rheumatoid arthritis based on PK/PD.
Shi-jia LIU ; Guo-liang DAI ; Bing-ting SUN ; Chang-yin LI ; Lei WU ; Ma SHI-TANG ; Wen-zheng JU ; Heng-shan TAN ; Hai-yan FU
China Journal of Chinese Materia Medica 2015;40(2):334-338
To observe the serum samples and the anti-inflammatory effect of Tripterygium wilfordii in treating RA by using the pharmacokinetic-pharmacodynamic model, make a correlation analysis on concentration-time and effect-time curves, and explore RORγt, IL-17, STAT3, IL-6 mRNA transcriptional levels in rats by PCR. Methotrexate, tripterine and high-dose T. wilfordii could down-regulate RORγt, IL-17, STAT3, IL-6 mRNA transcriptional levels in AA rat lymph nodes. The study on PK-PD model showed correlations between inflammatory factors and blood concentration of T. wilfordii. T. wilfordii and its main active constituent tripterine could show the inflammatory effect and treat RA by inhibiting IL-17 cytokine.
Animals
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Arthritis, Rheumatoid
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drug therapy
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immunology
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Biomarkers
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Female
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Interleukin-17
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antagonists & inhibitors
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genetics
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Interleukin-6
;
genetics
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Phytotherapy
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Rats
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Rats, Sprague-Dawley
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Tripterygium
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Triterpenes
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pharmacokinetics
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pharmacology
9.In vivo and in vitro anti-sepsis effects of physcion 8-O-β-glucopyranoside extracted from Rumex japonicus.
Wei-Jun FU ; Jian-Jun TANG ; Hui WANG ; Hong-Yun WEI ; Shu-Min CAI ; Zhen-Hua ZENG ; Hui CHEN ; Zhong-Qing CHEN
Chinese Journal of Natural Medicines (English Ed.) 2017;15(7):534-539
The present study was designed to investigate the anti-sepsis effects of physcion 8-O-β-glucopyranoside (POG) isolated from Rumex japonicas and explore its possible pharmacological mechanisms. POG was extracted from R. japonicas by bioactivity-guided isolation with the anti-sepsis agents. Survival analysis in septic mouse induced by LPS and heat-killed Escherichia coli were used to evaluate the protective effect of POG (40 mg·kg, i.p.) on sepsis. Cytokines including TNF-α, IL-1β and IL-6 in RAW 264.7 cells induced by LPS (100 ng·mL) were determined by ELISA. In addition, the proteins expressions of TLR2 and TLR4 were determined by Western blotting assay. Our results demonstrated that POG (40 mg·kg, i.p.) possessed significant protective activity on the endotoxemic mice. The POG treatment (20, 40, and 80 μg·mL) significantly decreased the TNF-α, IL-1β and IL-6 induced by LPS (P < 0.01) in a concentration-dependent manner. Furthermore, the TLR4 and TLR2 proteins were also down-regulated by POG at 20 (P < 0.01), 40 (P < 0.01), and 80 μg·mL (P < 0.01). The present study demonstrated that the POG extracted from R. japonicas possessed significant anti-sepsis effect on endotoxemic mice, and can be developed as a novel drug for treating sepsis in the future.
Animals
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Anti-Inflammatory Agents
;
administration & dosage
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Drugs, Chinese Herbal
;
administration & dosage
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Emodin
;
administration & dosage
;
analogs & derivatives
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Glucosides
;
administration & dosage
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Humans
;
Interleukin-1beta
;
genetics
;
immunology
;
Interleukin-6
;
genetics
;
immunology
;
Interleukin-8
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genetics
;
immunology
;
Macrophages
;
drug effects
;
immunology
;
Male
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Mice
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Mice, Inbred ICR
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RAW 264.7 Cells
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Rumex
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chemistry
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Sepsis
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drug therapy
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genetics
;
immunology
;
Tumor Necrosis Factor-alpha
;
genetics
;
immunology
10.Anti-inflammatory effect and mechanism of artemisinin and dihydroartemisinin.
Wan-ying YU ; Wei-juan KAN ; Peng-xia YU ; Min-min LI ; Ji-shuai SONG ; Feng ZHAO
China Journal of Chinese Materia Medica 2012;37(17):2618-2621
OBJECTIVETo study and compare the anti-inflammatory effect and molecular mechanism of artemisinin and dihydroartemisinin.
METHODMouse mononuclear macrophage RAW264.7 cells were stimulated to release inflammatory mediators such as TNF-alpha, IL-6 and NO, in order to assess the drugs' inhibitory effect on macrophage's release of above inflammatory mediators. The levels of TNF-alpha and IL-6 were determined by ELISA and the cytotoxicity was determined by MTT method. The protein expression of iNOS, COX-2 and beta-actin were tested by Western blot. The enzymatic activity of COX-2 was determined by colorimetric method.
RESULTDihydroartemisinin significantly inhibited LPS-induced release of TNF-alpha, IL-6 and NO from RAW264.7 in mice with the concentration range of 12.5 - 100 micromol x L(-1), and showed good dose dependence. Artemisinin only inhibited the IL-6 release to a certain extent.
CONCLUSIONDihydroartemisinin inhibits macrophages from releasing inflammatory factors TNF-alpha and IL-6 and inflammatory mediators NO by down-regulating iNOS protein. Artemisinin may help dihydroartemisinin to show its anti-inflammatory effect through metabolism.
Animals ; Anti-Inflammatory Agents ; pharmacology ; Artemisinins ; pharmacology ; Cell Line ; Gene Expression ; drug effects ; Inflammation Mediators ; immunology ; Interleukin-6 ; genetics ; immunology ; Macrophages ; drug effects ; immunology ; Mice ; Nitric Oxide ; immunology ; Tumor Necrosis Factor-alpha ; genetics ; immunology