1.The relativity between bone marrow mononuclear cells apoptosis and peripheral lymphocyte micronucleus in workers exposed to benzene.
Ling-li YE ; Miao-yong ZHU ; Hong-kang YE ; Jian-bo WU ; Zeng LIN ; Sheng-hui ZHANG ; Lei JIANG ; Ping CHEN ; Li-ming HU
Chinese Journal of Industrial Hygiene and Occupational Diseases 2004;22(2):140-141
Adult
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Apoptosis
;
drug effects
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Benzene
;
poisoning
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Biomarkers
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analysis
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Bone Marrow Cells
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drug effects
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Female
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Flow Cytometry
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Humans
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Leukocytes, Mononuclear
;
drug effects
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Lymphocytes
;
drug effects
;
metabolism
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Male
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Micronuclei, Chromosome-Defective
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drug effects
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metabolism
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Occupational Exposure
;
analysis
2.Effect of astragaloside against the oxidative damage on endothelial cells.
Kang-ting JI ; Ji-fei TANG ; Jun-de CHAI
Chinese Journal of Integrated Traditional and Western Medicine 2011;31(6):807-810
OBJECTIVETo observe the effect of astragaloside on oxidative low-density lipoprotein (Ox-LDL) mediated oxidative damage of endothelial progenitor cells (EPCs).
METHODSPeripheral blood mononuclear cells(PBMCs) were isolated by Ficoll density gradient centrifugation, and EPCs were identified by flow cytometry. Adherent cells were collected after seven-day incubation and randomly divided into the normal control group, the Ox-LDL group (as the model group, at the dose of 100 microg/mL), the low, middle, and high astragaloside groups (with 100 microg/mL Ox-LDL plus 2, 10, and 50 microg/mL astragaloside). Twenty-four h later, the proliferation and adhesion capabilities of EPCs were observed using MTT colorimetry and the adhesion capability detection. Levels of superoxide dismutase (SOD) and malonaldehyde (MDA) in the cell supernate of each group were determined.
RESULTSAfter Ox-LDL damage, the proliferative and adhesive capacities of EPCs were significantly injured (53 +/- 8 vs 42 +/- 6, 0.49 +/- 0.12 vs 0.37 +/- 0.02, both P<0.05). The SOD content obviously decreased (21.95 +/- 1.43 vs 14.76 +/- 3.99, P<0.01), the MDA content obviously increased (3.72 +/- 0.30 vs 5.57 +/- 0.64, P<0.01). After intervened by astragaloside for 24 h, the proliferative and adhesive capacities of EPCs were significantly improved. The SOD contents of astragaloside intervention groups were obviously improved and the MDA content obviously lowered.
CONCLUSIONSAstragaloside showed significant protection on Ox-LDL damaged EPCs. Its mechanism might be correlated with antioxidative damage.
Cells, Cultured ; Endothelial Cells ; cytology ; drug effects ; metabolism ; Humans ; Leukocytes, Mononuclear ; metabolism ; Lipoproteins, LDL ; metabolism ; Malondialdehyde ; metabolism ; Oxidation-Reduction ; Oxidative Stress ; drug effects ; Saponins ; pharmacology ; Stem Cells ; cytology ; drug effects ; metabolism ; Triterpenes ; pharmacology
3.Effect of spearmint oil on lipopolysaccharide induced emphysema-like changes and expression of matrix metalloproteinase-9.
Junbo LIU ; Yan WANG ; Fadi TANG ; Chenxi YU ; Mengshan HUANG ; Xiaojing ZHAO ; Youfa ZHU
China Journal of Chinese Materia Medica 2011;36(8):1054-1059
OBJECTIVETo investigate the effect of spearmint oil on emphysema-like changes and the expression of tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta(IL-1beta), matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-9) in lipopolysaccharide (LPS) treated rats.
METHODEmphysematous changes model was induced by intratracheal instillation of LPS once a week for up to 8 weeks in rats. Rats were divided into control, dexamethasone (0.3 mg x kg(-1)), and spearmint oil (10, 30,100 mg x kg(-1)) groups. Each group was treated with saline, dexamethasone, and spearmint of oil respectively for 4 weeks. Then total and different white blood cell counts in bronchoalveolar lavage fluid(BALF) were carried out. The pathologic changes of lung tissue such as alveolar structure, airway inflammation, and goblet cell metaplasia were observed by HE and AB-PAS staining. Expression of TNF-alpha, IL-1beta, TIMP-1 and MMP-9 were measured.
RESULTBoth spearmint and dexamethasone decreased the destruction of pulmonary alveolus. The total and different white blood cell counts in BALF including neutrophile and lymphocyte of spearmint oil 100 mg x kg(-1) and dexamethasone group were significantly reduced, and the goblet cell metaplasia was also inhibited. Dexamethasone had inhibitory effect on the expression of TNF-alpha, IL-1beta, TIMP-1 and MMP-9. Spearmint oil 30, 100 mg x kg(-1) significantly reduced TNF-alpha and IL-1beta respectively. Spearmint oil 10, 30 and 100 mg x kg(-1) had no effect on the expression of TIMP-1, but could decrease the expression of MMP-9 significantly in lung tissues.
CONCLUSIONSpearmint oil has protective effect on rats with emphysematous changes, since it improves alveolar destruction, pulmonary inflammation, and goblet cell metaplasia. The mechanism may include reducing TNF-alpha, IL-1beta content and inhibiting overexpression of matrix metalloproteinase-9 in lung tissues.
Animals ; Azo Compounds ; pharmacology ; Bronchoalveolar Lavage Fluid ; cytology ; Goblet Cells ; drug effects ; Interleukin-1beta ; drug effects ; metabolism ; Leukocytes ; drug effects ; metabolism ; Lipopolysaccharides ; Lymphocytes ; drug effects ; metabolism ; Matrix Metalloproteinase 9 ; drug effects ; metabolism ; Mentha spicata ; chemistry ; Metaplasia ; Monocytes ; drug effects ; metabolism ; Neutrophils ; drug effects ; metabolism ; Phytotherapy ; Plant Oils ; therapeutic use ; Pulmonary Emphysema ; chemically induced ; drug therapy ; enzymology ; pathology ; Rats ; Respiratory System ; drug effects ; pathology ; Tissue Inhibitor of Metalloproteinase-1 ; drug effects ; metabolism ; Tumor Necrosis Factor-alpha ; drug effects ; metabolism
4.Mechanism of glutamine downregulates the cytokine expression in lipopolysaccharide-stimulated human peripheral blood mononuclear cells.
Li-ming WANG ; Li-ya PAN ; Feng ZHANG ; Xin-ying WANG ; Jie-shou LI
Chinese Journal of Surgery 2009;47(20):1578-1580
OBJECTIVETo investigate the mechanism that glutamine (Gln) downregulates the cytokine expression in lipopolysaccharide (LPS)-stimulated human peripheral blood mononuclear cells (PMBCs).
METHODSPMBCs were extracted from healthy volunteer by density gradient centrifugation, the cells were divided into two parts. The first part of PMBCs was pretreated with Gln of the concentration of 0, 8, 15 mmol/L for 0.5 h and 2.0 h respectively, then stimulated by LPS for 4.0 h. Cells and supernatants were collected. The second part of PBMCs was divided into group A, B and C. Group A and B were pretreated with HSP70 blocker (Quercetin) for 1.0 h, then were stimulated by LPS for 4.0 h. Cells and supernatants were also collected. The release of TNF-alpha and IL-10 was analyzed via enzyme-linked immunosorbent assay (ELISA) and HSP70 via Western Blot. In this experiment, the effect of Quercetin on TNF-alpha, IL-10 and HSP70 expression in human PBMCs was assessed.
RESULTSGln led to an increase in HSP70 expression, and decreased TNF-alpha, IL-10 release at 4.0 h after LPS stimulation when 8 mmol/L glutamine pretreated for 0.5 h and 2.0 h, 15 mmol/L glutamine pretreated for 0.5 h (P < 0.05). The expression level of HSP70 was significantly decreased, however, the expression of TNF-alpha and IL-10 was enhanced in Quercetin group (P < 0.05).
CONCLUSIONThe effect of glutamine attenuating cytokine release in PBMCs is related to the enhancement of HSP70 expression.
Cells, Cultured ; Cytokines ; metabolism ; Glutamine ; pharmacology ; HSP70 Heat-Shock Proteins ; metabolism ; Humans ; Interleukin-10 ; metabolism ; Leukocytes, Mononuclear ; drug effects ; metabolism ; Lipopolysaccharides ; pharmacology ; Tumor Necrosis Factor-alpha ; metabolism
5.LRRC25 plays a key role in all-trans retinoic acid-induced granulocytic differentiation as a novel potential leukocyte differentiation antigen.
Weili LIU ; Ting LI ; Pingzhang WANG ; Wanchang LIU ; Fujun LIU ; Xiaoning MO ; Zhengyang LIU ; Quansheng SONG ; Ping LV ; Guorui RUAN ; Wenling HAN
Protein & Cell 2018;9(9):785-798
Leukocyte differentiation antigens (LDAs) play important roles in the immune system, by serving as surface markers and participating in multiple biological activities, such as recognizing pathogens, mediating membrane signals, interacting with other cells or systems, and regulating cell differentiation and activation. Data mining is a powerful tool used to identify novel LDAs from whole genome. LRRC25 (leucine rich repeat-containing 25) was predicted to have a role in the function of myeloid cells by a large-scale "omics" data analysis. Further experimental validation showed that LRRC25 is highly expressed in primary myeloid cells, such as granulocytes and monocytes, and lowly/intermediately expressed in B cells, but not in T cells and almost all NK cells. It was down-regulated in multiple acute myeloid leukemia (AML) cell lines and bone marrow cells of AML patients and up-regulated after all-trans retinoic acid (ATRA)-mediated granulocytic differentiation in AML cell lines and acute promyelocytic leukemia (APL; AML-M3, FAB classification) cells. Localization analysis showed that LRRC25 is a type I transmembrane molecule. Although ectopic LRRC25 did not promote spontaneous differentiation of NB4 cells, knockdown of LRRC25 by siRNA or shRNA and knockout of LRRC25 by the CRISPR-Cas9 system attenuated ATRA-induced terminal granulocytic differentiation, and restoration of LRRC25 in knockout cells could rescue ATRA-induced granulocytic differentiation. Therefore, LRRC25, a potential leukocyte differentiation antigen, is a key regulator of ATRA-induced granulocytic differentiation.
Antigens, Differentiation
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immunology
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metabolism
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Cell Differentiation
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drug effects
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Cell Line, Tumor
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Granulocytes
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cytology
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drug effects
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immunology
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metabolism
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Humans
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Leukocytes
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cytology
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drug effects
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immunology
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metabolism
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Membrane Proteins
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antagonists & inhibitors
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immunology
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metabolism
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RNA, Small Interfering
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pharmacology
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Tretinoin
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pharmacology
6.Effect of Lang-Chuang-Ding Decoction () on DNA Methylation of CD70 Gene Promoter in Peripheral Blood Mononuclear Cells of Female Patients with Systemic Lupus Erythematosus.
Jing SUN ; Tie-Juan SHAO ; Dong-Yu ZHANG ; Xue-Qin HUANG ; Zhi-Jun XIE ; Cheng-Ping WEN
Chinese journal of integrative medicine 2018;24(5):348-352
OBJECTIVETo investigate the effect of Lang-chuang-ding Decoction (, LCD) on the expression of DNA methylation of CD70 gene promoter in peripheral blood mononuclear cells (PBMCs) of females with systemic lupus erythematosus (SLE).
METHODSPBMCs isolated from female patients with SLE or healthy donors were cultured and treated with LCD medicated serum or normal serum for 24 or 48 h. The mRNA expressions of CD70 gene in PBMCs were detected by reverse transcription polymerase chain reaction (PCR); the DNA methylation of the CD70 gene promoter region was detected by methylation-specific PCR.
RESULTSAfter treated with medicated serum for 48 h, the mRNA expression levels of CD70 in PBMCs of SLE patients were signifificantly higher than those of healthy donors (P<0.05); the DNA methylation levels of CD70 promoter region in PBMCs of SLE patients treated with medicated serum for 48 h were signifificantly higher than those treated with fetal bovine serum (P<0.01).
CONCLUSIONLCD could inhibit CD70 gene expression in PBMCs of SLE patients by promoting the DNA methylation of CD70 gene promoter.
Adult ; CD27 Ligand ; genetics ; metabolism ; DNA Methylation ; drug effects ; genetics ; Drugs, Chinese Herbal ; pharmacology ; Female ; Gene Expression Regulation ; drug effects ; Humans ; Leukocytes, Mononuclear ; drug effects ; metabolism ; Lupus Erythematosus, Systemic ; drug therapy ; genetics ; Promoter Regions, Genetic ; RNA, Messenger ; genetics ; metabolism
7.Clinical study of correlation between the expression of ICBP90 and hematopoietic suppression in patients with chronic benzene poisoning.
Sheng LUO ; Xiao-yan HUANG ; Qian HU ; Xu-dong HU ; Chong-yun XING ; Kang YU
Chinese Journal of Industrial Hygiene and Occupational Diseases 2012;30(8):571-574
OBJECTIVETo observe the change of ICBP90 expression in patients with chronic benzene poisoning and explore the correlation between the expression of ICBP90 and benzene-induced hematotoxicity.
METHODSThe bone marrow samples were from 13 chronic benzene poisoning cases with hematopoietic suppression, 11 chronic benzene poisoning cases with hematopoietic regeneration and 10 controls. Western-blot was applied to detect the ICBP90 expression in bone marrow mononuclear cells (BMNCs). The correlation between ICBP90 expression and hematopoietic suppression in patients with chronic benzene poisoning was analyzed.
RESULTSThe ICBP90 expression of BMNCs in 13 chronic benzene poisoning cases with hematopoietic suppression was significantly lower than that in controls (P < 0.01). The ICBP90 expression of BMNCs in 11 chronic benzene poisoning cases with hematopoietic regeneration was significantly higher than those in controls and 13 chronic benzene poisoning cases with hematopoietic suppression (P < 0.05 or P < 0.01), respectively. There were good correlations between the expression of ICBP90 and white blood cell and platelet counts in patients with chronic benzene poisoning (r(1) = 0.555,P = 0.006; r(2) = 0.854,P < 0.01).
CONCLUSIONThe ICBP90 expression of BMNCs in the chronic benzene poisoning cases with hematopoietic suppression decreased significantly, and the ICBP90 expression of BMNCs in the chronic benzene poisoning cases with hematopoietic regeneration increased significantly. There was good correlation between hematopoietic suppression and ICBP90 expression in patients with chronic benzene poisoning.
Adult ; Benzene ; poisoning ; Blood Platelets ; metabolism ; Bone Marrow Cells ; metabolism ; CCAAT-Enhancer-Binding Proteins ; metabolism ; Case-Control Studies ; Female ; Hematopoiesis ; drug effects ; Humans ; Leukocytes ; metabolism ; Male ; Young Adult
8.Effects of rare earth compounds on human peripheral mononuclear cell telomerase and apoptosis.
Li YU ; Yu-Cheng DAI ; Zhao-Kang YUAN ; Jie LI
Chinese Journal of Preventive Medicine 2004;38(4):248-251
OBJECTIVETo study the effects of rare earth exposure on human telomerase and apoptosis of human peripheral mononuclear cells (PBMNs).
METHODSRare earth mine lot in Xunwu county, the biggest ion absorptive rare earth mine lot of China, was selected as the study site. Another village of Xunwu county, with comparable geological structure and social environment was selected as the control site. Thirty healthy adults were randomly selected from the study site as exposure group and another 30 healthy adults randomly selected from the control site as control group. The blood content of 15 rare earth elements, including La, Ce, Pr, Nd, Sm, Eu, Gd, Tb, Dy, Ho, Er, Tm, Yb, Lu and Y, were determined by inductive coupled plasma-source mass spectrometry (ICP-MS). The total contents of rare earth elements in the blood were calculated. The TRAP and FCM assays were carried out to analyse the telomerase and apoptosis of human PBMNCs respectively.
RESULTSIn the exposure group, the concentration of La, Ce, Dy and Y were significantly higher (P<0.001), and Pr, Nd, Sm, Gd and Yb were higher than those in the control group (P<0.05). The total content of rare earth in the blood of exposure group showed significant difference compared with control group (P<0.001). Telomerase activity in PBMNs of the exposure group was higher than that in the control group (P<0.05); there were 11 adults in the exposure group (30 adults) and 5 adults in control group (30 adults) showed positive telomerase activity. The average age of the exposure group was (38.69 +/- 8.02) years-old, while the control group was (40.45 +/- 9.02) years-old (P >0.05). It was found that there was a significant relationship between telomerase activity and the total content of rare earth elements (P <0.01). 3. The proportion of apoptosis was not different between the two groups (P >0.05), but the cells in the S-phase and G2-M phase were increased (P <0.01) in the exposed group.
CONCLUSIONThe telomerase activity of PBMNs in the rare earth elements exposed group was higher than that of the control group, and there is no effect on apoptotic rate of PBMNs, but may promote the diploid DNA replication, and increase the percentage of G2/M and S phase cells.
Apoptosis ; drug effects ; Environmental Exposure ; adverse effects ; Female ; Humans ; Leukocytes, Mononuclear ; cytology ; enzymology ; Male ; Metals, Rare Earth ; adverse effects ; analysis ; Telomerase ; metabolism
9.Enhanced sensitivity of leukemia cell line KG-1a to activated immune cell-mediated cytolysis after treated with resveratrol.
Liangshan HU ; Huawen YANG ; Lihua LI ; Zhihong ZHANG ; Xiaolin FANG ; Donglin CAO
Chinese Journal of Hematology 2014;35(7):645-649
OBJECTIVETo explore the enhanced sensitivity of leukemia cell line KG-1a to activated immune cell-mediated cytolysis after treated with resveratrol.
METHODSThe value of 50% inhibition concentration (IC₅₀) for KG-1a by resveratrol was analyzed using trypan blue staining. Peripheral blood mononuclear cells were separated, and then activated by interleukin (IL)-2 and IL-15. The sensitivity of KG-1a treated with and without resveratrol to activated immune cell-mediated cytolysis was assayed by lactate dehydrogenase (LDH) -releasing assay. The expression of tumor necrosis factor related apoptosis inducing ligand (TRAIL) on the surface of activated immune cells and its receptors (DR4/5 and DcR1/2) on the surface of KG-1a were detected by flow cytometry.
RESULTSResveratrol could inhibit the proliferation of KG-1a and IC50 at 24 h was 25 mmol/L. At a ratio of 10:1 or 20:1 between effect and target, the cytolytic rates of treated KG-1a by activated immune cells were (55.80 ± 10.88)% and (72.31 ± 13.06)%, significantly higher than (24.96 ± 9.25)% and (37.93 ± 5.21)% of untreated KG-1a (P<0.05). The expression of DR5 on the surface of KG-1a treated with resveratrol was (9.05 ± 3.57)%, significantly higher than (3.11 ± 0.54)% of untreated KG-1a (P<0.05). Conversely, the expression of DcR1 on the surface of treated KG-1a was (13.23 ± 3.56)%, lower than (53.75 ± 10.51)% of KG-1a (P<0.05). When TRAIL pathway on the surface of activated immune cells was blocked, the cytolytic rates of treated KG-1a were (35.97 ± 6.36)% and (49.80 ± 10.68)%, significantly lower than (52.92 ± 6.98)% and (70.73 ± 9.79)% of untreated KG-1a (P<0.05) at the same ratio of effector and target.
CONCLUSIONResveratrol could enhance cytolytic sensitivity of KG-1a by activated immune cells through TRAIL pathway.
Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Leukemia ; metabolism ; pathology ; Leukocytes, Mononuclear ; drug effects ; immunology ; metabolism ; Male ; Receptors, TNF-Related Apoptosis-Inducing Ligand ; metabolism ; Receptors, Tumor Necrosis Factor, Member 10c ; metabolism ; Stilbenes ; pharmacology ; TNF-Related Apoptosis-Inducing Ligand ; metabolism
10.Effects of various inducers on the expression of P2X7 receptor in human peripheral blood mononuclear cells.
Xiu-Jun ZHANG ; Guo-Guang ZHENG ; Xiao-Tong MA ; Yong-Min LIN ; Yu-Hua SONG ; Ke-Fu WU
Acta Physiologica Sinica 2005;57(2):193-198
Regulation of P2X7 receptor expression is of interest because activation of this receptor by extracellular ATP triggers a wide variety of cell functions in leukocytes. However, its expression and modulation in human peripheral blood mononuclear cells (PBMC) and monocytes remain unclear. RT-PCR was used to detect the constitutive level of P2X7 receptor and the levels upon stimulation with bacteria, bacterial product, mitogen and various cytokines in human PBMC and monocytes. P2X7 receptor mRNA was detected in PBMC and monocytes. P2X7 receptor expression in PBMC was up-regulated by interleukin-2, -4, -6 (IL-2, IL-4, IL-6) tumour necrosis factor-alpha (TNF-alpha), lipopolysaccharide (LPS) and heat-inactivated Staphylococcus aureus Cowan strain I (SAC). However, interferon-gamma (IFN-gamma), granulocyte-macrophage colony-stimulating factor (GM-CSF), macrophage colony-stimulating factor (M-CSF) and phytohemagglutinin-M (PHA-M) had little effect on the expression of P2X7 receptor. Furthermore, LPS and M-CSF could up-regulate P2X7 receptor expression in monocytes, while IFN-gamma, TNF-alpha and GM-CSF had weak effects, but pretreatment with these inducers could not further enhance LPS-stimulated P2X7 receptor expression in monocytes. The results obtained demonstrate that inflammatory stimuli drive P2X7 expression, thus supporting the hypothesis that P2X7 receptor may play a role in the inflammatory responses against bacteria infection, which need further verification.
Humans
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Interleukin-2
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physiology
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Interleukin-4
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physiology
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Leukocytes, Mononuclear
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drug effects
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metabolism
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RNA, Messenger
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genetics
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metabolism
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Receptors, Purinergic P2X7
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genetics
;
metabolism
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Tumor Necrosis Factor-alpha
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physiology