OBJECTIVETo observe the effect of Chinese herbs for supplementing Shen and strengthening bone (HB) on myelogenic osteoclasts formation, and gene expression of interleukin-6 (IL-6), IL-6 receptor (IL-6R) and gp130 in bone marrow.

METHODSSeventy-two healthy female SD rats of 3 months, were randomly divided into three groups, 24 in the sham-operated group (A), 24 in the ovariectomized group (B) and 24 in the after ovariectomy HB treated group (C). Bone marrow cells of 6 rats from each group were respectively collected and cultured at four time points (2nd, 4th, 6th and 12th weeks after operation). After 6 days of culture, the bone marrow cells were differentiated by Wright-Giemsa stain and TRAP stain, and total RNA in them was extracted by TRIZOL.

RESULTSBeginning from the 2nd week, the osteoclasts formation in Group B was higher than that in Group A (P < 0.05), and IL-6, IL-6R gene expression significantly increased in Group B (P < 0.05 or P < 0.01). These changes reached the peak in the 4th to 6th week, with the level maintained to the 12th week. As for comparison of Group B and C, the above-mentioned changes were significantly weakened in the latter (P < 0.05 or P < 0.01). No significant change of gp130 gene expression revealed in the whole course in either group.

CONCLUSIONHB could inhibit the myelogenic osteoclasts formation in ovariectomized rats, this effect may be correlated with, partially at least, its inhibitory effect on the over-expressed IL-6 and IL-6R gene expression in myelocytes after ovariectomy.

Animals ; Antigens, CD ; biosynthesis ; genetics ; Bone Marrow ; metabolism ; Cell Division ; drug effects ; Cells, Cultured ; Cytokine Receptor gp130 ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Female ; Granulocyte Precursor Cells ; metabolism ; Interleukin-6 ; biosynthesis ; genetics ; Isoflavones ; pharmacology ; Membrane Glycoproteins ; biosynthesis ; genetics ; Osteoblasts ; pathology ; Osteoporosis ; metabolism ; pathology ; Ovariectomy ; RNA ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, Interleukin-6 ; biosynthesis ; genetics

Purpose: IL-1 is a multifunctional proinflammatory cytokine. As the proliferative effects of IL-6 and IL-6 receptor expressions on prostatic cancer cells in response to IL-1 have not been determined, the effects of IL-1 on prostatic cancer cell lines were investigated. Materials and Methods: PC-3 and DU-145 cells were cultured in RPMI-1640 medium containing 10% fetal bovine serum. Cell cultures were supplemented with various concentrations of IL-1 (0, 1, 10, 20 and 40ng/ml), and the MMT growth assay performed. PC-3 and DU-145 cells were treated for 2, 4, 8, 12 and 24 h both with and without IL-1. IL-6 and IL-6 receptor (IL-6R) mRNA expressions were investigated using RT-PCR, and the IL-6 levels in cultured supernatant measured by ELISA. Results: The viability of both PC-3 and DU-145 cells decreased after IL-1 treatment (10, 20 and 40ng/mul). With 40ng/ml the IL-1, IL-6 and IL-6RmRNA expressions were lower in PC-3 cells, but unchanged in DU-145 cells, whereas the IL-6 protein production was higher in both PC-3 and DU-145 cells. Conclusions: IL-1 inhibited the proliferation of both PC-3 and DU145 cells. In the PC-3 cells, IL-1 decreased the expressions of IL-6 and IL-6R mRNA, but paradoxically increased the IL-6 production. In the DU-145 cells, IL-1 treatment did not affect the IL-6 or IL-6R mRNA expressions, but the IL-6 production was increased. This discrepancy between IL-1-induced IL-6 mRNA and protein production may be mediated by modification to the protein synthesis or an increased cellular excretion.
Cell Culture Techniques ; Cell Line ; Enzyme-Linked Immunosorbent Assay ; Interleukin-1 ; Interleukin-1beta* ; Interleukin-6* ; Interleukins* ; Prostatic Neoplasms* ; Receptors, Interleukin* ; Receptors, Interleukin-6 ; RNA, Messenger

Imaging Ellipsometry is one of recently developed optical surface-sensitive methods for the investigation of various aspects of biomolecules adsorption on solid surfaces and biomolecules interactions. It has advantages of high sensitivity to layer-thickness, big area of view, high sampling speed, and high lateral resolution. Compared with other solid phase methods such as enzyme linked immunosorbent assay, immunofluorescence and radioimmunoassay, imaging ellipsometry has the advantage of not involving any labelling of reactants and it is a relatively inexpensive method and easy to handle. In this report, the mono-layers of IL-6 and IL-6 receptor were visualized as well as their interaction.
Diagnostic Imaging ; instrumentation ; methods ; Humans ; Interleukin-6 ; metabolism ; Microscopy, Polarization ; methods ; Receptors, Interleukin-6 ; metabolism ; Surface Properties

OBJECTIVETo investigate the biological activity and molecular mechanism of interferon alpha (IFNalpha) on human myeloma cell line Sko-007.

METHODSThe effect of IFNalpha on the growth of Sko-007 cells was measured by MTT assay. Cells cycle distribution and the expression of two IL-6 receptor chains (IL-6R and gp130) on Sko-007 cell surface in the absence or presence of IFNalpha were monitored by FACS analysis. The activation state of protein kinase ERK, which is involved in Ras/MAPK signal transduction pathway mediating cell survival and proliferation, and the expression of anti-apoptotic Bcl-2 family proteins-Bcl-2, Bcl-x(L) and Mcl-1 in Sko-007 cells with or without IFNalpha were determined by immunoblot assay.

RESULTIFNalpha arrested Sko-007 cell cycle progression. After stimulation with IFNalpha, an obvious increase in G(0)/G(1) phase (41.1%-->84.1%) and decrease in S phase (57.1%-->13.3%) of Sko-007 cell cycle distribution can be observed. Moreover, the proliferation of Sko-007 cells was dramatically inhibited in the presence of IFNalpha, with a maximal inhibitory rate up to 88%. In addition, the expression of gp130 on cell surface, the activation of protein kinase ERK and the expression of Bcl-2 and Bcl-x(L) were all down-regualted in IFNalpha-stimulated Sko-007 cells.

CONCLUSIONThe inhibitory effect of IFNalpha on the proliferation of Sko-007 cells was mediated by gp130 down-regulation, degradation of Bcl-2 family anti-apoptotic proteins and inhibition of ERK activation.

Antigens, CD ; drug effects ; metabolism ; Cell Cycle ; drug effects ; Cell Division ; drug effects ; Cytokine Receptor gp130 ; Dose-Response Relationship, Drug ; Down-Regulation ; Enzyme Activation ; drug effects ; G1 Phase ; drug effects ; Humans ; Immunoblotting ; Interferon-alpha ; pharmacology ; Membrane Glycoproteins ; drug effects ; metabolism ; Mitogen-Activated Protein Kinases ; metabolism ; Multiple Myeloma ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Receptors, Interleukin-6 ; drug effects ; metabolism ; Resting Phase, Cell Cycle ; drug effects ; S Phase ; drug effects ; Tumor Cells, Cultured ; drug effects ; metabolism ; bcl-X Protein

OBJECTIVE: Schizophrenia is a disabling disorder of unknown aetiology, lacking definite diagnostic method and cure. A reliable biological marker of schizophrenia is highly demanded, for which traceable immune mediators in blood could be promising candidates. We aimed to gather the best findings of neuroinflammatory markers for first-episode psychosis (FEP). METHODS: We performed an extensive narrative review of online literature on inflammation-related markers found in human FEP patients only. RESULTS: Changes to cytokine levels have been increasingly reported in schizophrenia. The peripheral levels of IL-1 (or its receptor antagonist), soluble IL-2 receptor, IL-4, IL-6, IL-8, and TNF-α have been frequently reported as increased in FEP, in a suggestive continuum from high-risk stages for psychosis. Microglia and astrocytes establish the link between this immune signalling and the synthesis of noxious tryptophan catabolism products, that cause structural damage and directly hamper normal neurotransmission. Amongst these, only 3-hydroxykynurenine has been consistently described in the blood of FEP patients. CONCLUSION: Peripheral molecules stemming from brain inflammation might provide insightful biomarkers of schizophrenia, as early as FEP or even prodromal phases, although more time- and clinically-adjusted studies are essential for their validation.
Astrocytes ; Biomarkers ; Encephalitis ; Humans ; Interleukin-1 ; Interleukin-4 ; Interleukin-6 ; Interleukin-8 ; Metabolism ; Methods ; Microglia ; Polytetrafluoroethylene ; Psychotic Disorders ; Receptors, Interleukin-2 ; Schizophrenia ; Synaptic Transmission ; Tryptophan

Resistance training (RT) is associated with reduced risk of low grade inflammation related diseases, such as cardiovascular disease and type 2 diabetes. The majority of the data studying cytokines and exercise comes from endurance exercise. In contrast, evidence establishing a relationship between RT and inflammation is more limited. This review focuses on the cytokine responses both following an acute bout, and after chronic RT. In addition, the effect of RT on low grade systemic inflammation such as individuals at risk for type 2 diabetes is reviewed. Cytokines are secreted proteins that influence the survival, proliferation, and differentiation of immune cells and other organ systems. Cytokines function as intracellular signals and almost all cells in the body either secrete them or have cytokine receptors. Thus, understanding cytokine role in a specific physiological situation such as a bout of RT can be exceedingly complex. The overall effect of long term RT appears to ameliorate inflammation, but the specific effects on the inflammatory cytokine, tumor necrosis factor alpha are not clear, requiring further research. Furthermore, it is critical to differentiate between chronically and acute Interleukin-6 levels and its sources. The intensity of the RT and the characteristics of the training protocol may exert singular cytokine responses and as a result different adaptations to exercise. More research is needed in the area of RT in healthy populations, specifically sorting out gender and age RT acute responses. More importantly, studies are needed in obese individuals who are at high risk of developing low grade systemic inflammatory related diseases. Assuring adherence to the RT program is essential to get the benefits after overcoming the first acute RT responses. Hence RT could be an effective way to prevent, and delay low grade systemic inflammatory related diseases.
Cardiovascular Diseases ; Cytokines ; Inflammation ; Interleukin-6 ; Proteins ; Receptors, Cytokine ; Resistance Training ; Tumor Necrosis Factor-alpha

Prostate cancer is the most frequently diagnosed cancer. Although prostate tumors respond to androgen ablation therapy at an early stage, they often acquire the potential of androgen-independent growth. Elevated transcriptional activity of androgen receptor (AR) and/or signal transducer and activator of transcription-3 (STAT3) contributes to the proliferation of prostate cancer cells. In the present study, we examined the effect of resveratrol, a phytoalexin present in grapes, on the reporter gene activity of AR and STAT3 in human prostate cancer (LNCaP-FGC) cells stimulated with interleukin-6 (IL-6) and/or dihydrotestosterone (DHT). Our study revealed that resveratrol suppressed the growth of LNCaP-FGC cells in a time- and concentration-dependent manner. Whereas the AR transcriptional activity was induced by treatment with either IL-6 or DHT, the STAT3 transcriptional activity was induced only by treatment with IL-6 but not with DHT. Resveratrol significantly attenuated IL-6-induced STAT3 transcriptional activity, and DHT- or IL-6-induced AR transcriptional activity. Treatment of cells with DHT plus IL-6 significantly increased the AR transcriptional activity as compared to DHT or IL-6 treatment alone and resveratrol markedly diminished DHT plus IL-6-induced AR transcriptional activity. Furthermore, the production of prostate-specific antigen (PSA) was decreased by resveratrol in the DHT-, IL-6- or DHT plus IL-6-treated LNCaP-FGC cells. Taken together, the inhibitory effects of resveratrol on IL-6- and/or DHT-induced AR transcriptional activity in LNCaP prostate cancer cells are partly mediated through the suppression of STAT3 reporter gene activity, suggesting that resveratrol may be a promising therapeutic choice for the treatment of prostate cancer.
Dihydrotestosterone ; Genes, Reporter ; Humans ; Interleukin-6 ; Prostate ; Prostate-Specific Antigen ; Prostatic Neoplasms* ; Receptors, Androgen ; Transducers ; Vitis

OBJECTIVEThis study further explores the stromal cell-derived factor-1 (SDF-1)/chemokine receptor 4 (CXCR4) signaling axis mechanism in temporomandibular joint osteoarthritis (OA) by detecting the changes in CXCR4, interleukin (IL)-6, and collagen X expression in the ATDC5 cell line stimulated by the cyclic tensile strain and SDF-1.

METHODSInsulin-transferrin-selenium (ITS) was used to induce ATDC5 cells to differentiate into chondrocyte-like cells. After three weeks, the cells were divided into two groups: those with and without cyclic tensile strain. These groups were further divided into the negative control and SDF-1 groups. Strain force of 20% was applied. After 12 h, the total proteins were extracted from cells of the four groups, and Western blot analysis was used to detect the changes in CXCR4, IL-6, and collagen X expression.

RESULTSSDF-1 could enhance CXCR4, IL-6, and collagen X expressions in the chondrocytes, and 20% tensile strain force could further upregulate the three factors.

CONCLUSIONUnder abnormal tensile force, SDF-1 can upregulate its specific receptor CXCR4, thus increasing its-binding efficiency and resulting in the activation of the SDF-1/CXCR4 axis. This condition enhances the expressions of IL-6 and other inflammatory factors and directly damages to cartilage tissue. Such damage directly promotes chondrocyte hypertrophy, which enhances collagen X expression.

Cell Differentiation ; Cell Line ; Chemokine CXCL12 ; Collagen ; Humans ; Interleukin-6 ; Receptors, CXCR4 ; Signal Transduction ; Stromal Cells

BACKGROUND: Bone marrow stromal cells (BMSCs) express many cell surface molecules, which regulate the proliferation and differentiation of immune cells within the bone marrow. METHODS: To identify cell surface molecules, which can regulate cell proliferation through cell interaction, monoclonal antibodies (MoAbs) against BMSCs were produced. Among them, 1H8 MoAb, which recognized distinctly an 80 kDa protein, abolished myeloma cell proliferation that was induced by co-culturing with BMSCs. RESULTS: IL-6 gene expression was increased when myeloma or stromal cells were treated with 1H8 MoAb. In addition, the expression of IL-6 receptor and CD40 was up-regulated by 1H8 treatment, suggesting that the molecule recognized by 1H8 MoAb is involved in cell proliferation by modulating the expression of cell growth-related genes. Myeloma cells contain high levels of reactive oxygen species (ROS), which are related to gene expression and tumorigenesis. Treatment with 1H8 decreased the intracellular ROS level and increased PAG antioxidant gene concomitantly. Finally, 1H8 induced the tyrosine phosphorylation of several proteins in U266. CONCLUSION: Taken together, 1H8 MoAb recognized the cell surface molecule and triggered the intracellular signals, which led to modulate gene expression and cell proliferation.
Antibodies, Monoclonal ; Bone Marrow* ; Carcinogenesis ; Cell Communication ; Cell Proliferation ; Gene Expression ; Humans* ; Interleukin-6 ; Mesenchymal Stromal Cells* ; Phosphorylation ; Reactive Oxygen Species ; Receptors, Interleukin-6 ; Stromal Cells ; Tyrosine

OBJECTIVE: To characterize Stat activity in synovial tissue in rheumatoid arthritis (RA) in order to see if Stat molecule contributes to the pathogenesis of RA by regulatory expression of genes that play an important role in inflammation and tissue destruction. METHODS: Synovial tissue were obtained immediately after operative excision. Immuno-histochemistry was done with the antibodies for Stat 3 and Stat 5. Cells were stimulated with interleukin 6 (IL-6) and soluble interleukin 6 receptor (sIL-6R) or steroid using chambered slide. In supershift experiment, cell extracts were incubated with 0.5ng of 32P-labelled double-stranded oligonucleotide probe. Samples were resolved on 4.5% polyacrylamide gels, which was transferred to polyvinylidene fluoride membranes. Anti-phosphotyrosine Stat 3 antibody was used for Western blotting. RESULTS: Stat 3 was not shown on the synovial tissue section done by immuno-histochemistry. However, activated Stat 3 was expressed on cultured synovial cell stimulated with IL-6 and sIL-6R, and also with IL-6 and dexamethasone using chambered slide. In contrast to Stat 3, activated Stat 5 was expressed on the synovial tissue section, especially around blood vessel. CONCLUSION: Stat is activated in cultured synovial cells as shown in other immune associated cells, and IL-6 is the strong activator of Stat 3. Further analysis of the regulation of Stats in synovitis and the role of Stats in driving synovial inflammation will yield insight into the pathogenesis of RA and the development of novel therapeutic modality.
Antibodies ; Arthritis, Rheumatoid* ; Blood Vessels ; Blotting, Western ; Cell Extracts ; Dexamethasone ; Fluorides ; Gels ; Inflammation ; Interleukin-6 ; Membranes ; Receptors, Interleukin-6 ; Signal Transduction ; Synovitis