The aim of this study was to explore the effects of the stromal cell-derived factor (SDF-1) and chemokine receptors (CXCR4) on chemotaxis of cord blood AC133(+) cells. The optimal SDF-1 concentration was determined in Transwell System. The cell migration was calculated from the number of cells passing through polycarbonate membrane with 8 microm pore. The expressions of CXCR4 in fresh and cultured cord blood AC133(+) cells were analyzed by flow cytometry with two-color direct immunofluorescence. The results showed that the chemotactic rate of fresh cord blood AC133(+) cells increased along with increasing concentrations of SDF-1, however, it tended to be stable when the concentration of SDF-1 reached 150 ng/ml. There was no difference in the chemotactic rate of cord blood AC133(+) cells between the group with SDF-1 adding CXCR4-blocking antibody and the group without SDF-1. When AC133(+) cells were cultured in vitro with hemopoietic growth factors, the expression of CXCR4 increased at the early stage, but decreased gradually along with time extending. In conclusion, there was correlation between the chemotactic rate of AC133(+) cells and the expression of chemokine receptor CXCR4.
Cell Line ; Chemokine CXCL12 ; pharmacology ; Chemotaxis ; Fetal Blood ; cytology ; Humans ; Receptors, CXCR4 ; metabolism ; Stromal Cells ; metabolism

OBJECTIVES: This study aimed to investigate how naringenin (Nar) affected the anti-inflammatory, vascula-rization, and osteogenesis differentiation of human periodontal ligament stem cells (hPDLSCs) stimulated by lipopolysaccharide (LPS) and to preliminarily explore the underlying mechanism. METHODS: Cell-counting kit-8 (CCK8), cell scratch test, and Transwell assay were used to investigate the proliferation and migratory capabilities of hPDLSCs. Alkaline phosphatase (ALP) staining, alizarin red staining, lumen-formation assay, enzyme-linked immunosorbent assay, quantitative timed polymerase chain reaction, and Western blot were used to measure the expression of osteopontin (OPN), Runt-related transcription factor 2 (RUNX2), vascular endothlial growth factor (VEGF), basic fibroblast growth factor (bFGF), von Willebrand factor (vWF), tumor necrosis factor-α (TNF-α), and interleukin (IL)-6. RESULTS: We observed that 10 μmol/L Nar could attenuate the inflammatory response of hPDLSCs stimulated by 10 μg/mL LPS and promoted their proliferation, migration, and vascularization differentiation. Furthermore, 0.1 μmol/L Nar could effectively restore the osteogenic differentiation of inflammatory hPDLSCs. The effects of Nar's anti-inflammatory and promotion of osteogenic differentiation significantly decreased and inflammatory vascularization differentiation increased after adding AMD3100 (a specific CXCR4 inhibitor). CONCLUSIONS Nar demonstrated the ability to promote the anti-inflammatory, vascularization, and osteogenic effects of hPDLSCs stimulated by LPS, and the ability was associated with the stromal cell-derived factor/C-X-C motif chemokine receptor 4 signaling axis.
Humans ; Anti-Inflammatory Agents/pharmacology* ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Chemokine CXCL12 ; Lipopolysaccharides/pharmacology* ; Osteogenesis ; Periodontal Ligament/metabolism* ; Receptors, Chemokine/metabolism* ; Stem Cells ; Interleukin-8/metabolism*

OBJECTIVE: To investigate the role of IL-17A in promoting the activation of lung fibroblasts and the secretion of chemokine CXCL12, and to explore the possible mechanism. METHODS: Lung tissues of BALB/c mice were collected after intraperitoneal injection of recombinant mouse IL-17A (rmIL-17A). Real-time RT-PCR and Western blotting were used to detect the expression levels of α-smooth muscle actin (α-SMA) and collagen I in lung tissues, and immunohistochemical staining and real-time RT-PCR were used to determine the expression of CXCL12. Normal mouse primary lung fibroblasts were isolated and cultured, and identified by immunofluorescence staining with optical microscopy. Cells and supernatant of culture medium were collected after stimulation with rmIL-17A at different concentrations. mRNA levels of α-SMA, collagen I, and CXCL12 in the cells were determined by real-time RT-PCR, and the levels of collagen I and CXCL12 in the supernatant of culture medium were determined by ELISA. RESULTS: The mRNA and protein levels of α-SMA and collagen I in the lung tissue of mice injected with rmIL-17A were significantly increased compared with the control group (all CONCLUSIONS s: IL-17A can promote the activation of lung fibroblasts and translation into myofibroblast. The secretion of collagen is increased, which promote the deposition of extracullular matrix, and leads to the occurrence and development of lung fibrosis. CXCL12, a chemokine secreted by activated fibroblasts, may be involved in this process.
Actins/genetics* ; Animals ; Cells, Cultured ; Chemokine CXCL12/metabolism* ; Fibroblasts/metabolism* ; Interleukin-17/pharmacology* ; Lung/metabolism* ; Mice ; Mice, Inbred BALB C

OBJECTIVETo investigate the effects of shikonin on the proliferation, expression of CXCR4 and the migratory responses to CXCL12 in colorectal carcinoma cell line SW480.

METHODSThe proliferation of SW480 cells was assessed by MTT assay. Cell surface expression of CXCR4 was determined by flow cytometry. The migratory ability was determined by Transwell.

RESULTSShikonin inhibited the proliferation of SW480 cells in time- and concentration-dependent manner. The expression rate of CXCR4 in SW480 cells was 99.1%. After application of shikonin 0.01 micromol/L, 0.1 micromol/L and 1.0 micromol/L for 24 h, the expression rate of CXCR4 decreased to 76.0%, 59.1% and 35.5% respectively (F=1098.041, P <0.001), and the CXCL12-induced SW480 cell migratory inhibition rate was 25.2%, 38.5% and 55.7% respectively (F=48.970, P <0.001).

CONCLUSIONBesides having inhibiting tumor cell proliferation effect, Shikonin may also play a role in anti-metastasis via down-regulating the expression of CXCR4 and reducing the CXCL12-induced migratory response in colorectal carcinoma cell.

Cell Line, Tumor ; Cell Proliferation ; drug effects ; Chemokine CXCL12 ; metabolism ; Down-Regulation ; Humans ; Naphthoquinones ; pharmacology ; Receptors, CXCR4 ; metabolism

B-1 cells, which constitute a predominant lymphocyte subset in serosal cavities and produce most of natural antibodies, are subdivided into the CD5+ B-1a and CD5- B-1b cell subpopulations, but the differential roles of B-1a and B-1b cells are not well understood. We report that B-1a cells preferentially migrate out of the peritoneal cavity and upregulate the expression of CXCR4 with heightened sensitivity to CXCL12 and CXCL13 upon LPS treatment compared to B-1b and B-2 cells. Whereas B-1a cells were slightly more abundant than B-1b and B-2 cells in the homeostatic condition, the number of B-1a cells preferentially decreased 48 hr after LPS treatment. The decrease in the peritoneal B-1a cell number was accompanied with increased migration of B-1a cells toward CXCL-12 and CXCL-13 in in vitro transmigration assay using peritoneal B cells from LPS treated mice. The expression level of CXCR4, but not of CXCR5, was also more prominently increased in B-1a cells upon LPS stimulation. LPS-stimulated B-1a cells did not accumulate in omental milky spots in contrast to B-2 cells. These results suggest that B-1a cells actively migrate out of the peritoneal cavity through the regulation of the migratory responsiveness to chemokines and actively participate in systemic immune responses.
Adjuvants, Immunologic/pharmacology ; Animals ; B-Lymphocytes/cytology/*drug effects/immunology ; Cell Movement ; Cells, Cultured ; Chemokine CXCL12/metabolism/*pharmacology ; Chemokine CXCL13/metabolism/pharmacology ; Lipopolysaccharides/*pharmacology ; Mice ; Mice, Inbred C57BL ; Peritoneal Cavity/cytology ; Receptors, CXCR4/*metabolism ; Up-Regulation

OBJECTIVETo study the effect of total saponins from Rhizoma Dioscoreae nipponicae (RDN) on the expression of stroma cell derived factor 1 (SDF1) and IKB kinase (IKK) in rIL-1beta induced fibroblast-like synoviocytes (FLS).

METHODSFLS were primarily cultured and the 3rd generation log phase growth FLS were divided into the normal control group, the model group, and the medication group. 10 microg/L rIL-1beta was used to induce the proliferation of FLS in the model group.10 microg/L rIL-1beta and 100 microg/L RDN were administered to co-incubate FLS in the medication group. No treatment was given to FLS in the normal control group. Expression levels of SDF1 and IkapaB kinase proteins (p-IKK) were detected using Western blot.

RESULTSExpression levels of SDF1 and p-IKK increased significantly higher in the model group than in the normal control group (P<0.01). Compared with the model group, expression levels of SDF-1 and p-IKK significantly decreased in the medication group (P <0.01).

CONCLUSIONSTotal saponins from RDN could inhibit the activation of both SDF1 and p-IKK. It might further regulate the expression of IKB kinase by regulating the expression of SDF1.

Cells, Cultured ; Chemokine CXCL12 ; metabolism ; Dioscorea ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Epithelial Cells ; Fibroblasts ; Humans ; Interleukin-1beta ; metabolism ; Saponins ; metabolism ; Synovial Membrane ; metabolism

OBJECTIVETo investigate the expression of stromal cell-derived factor-1 (SDF-1) in human stem cells from apical papilla (SCAP), and to evaluate the effect of lipopolysaccharide (LPS) on SDF-1 expression by SCAP.

METHODSSCAP were isolated from dental papilla of human immature third molars. The expression of SDF-1 was evaluated by reverse transcription-PCR (RT-PCR). After SCAP being exposed to different concentrations (0.1, 1.0, 10 mg/L) of LPS for 24 and 48 h, the effect of LPS on cell proliferation and gene expression of SDF-1 was investigated by cell counting kit-8 and real-time PCR respectively, while cells without LPS stimulation were considered as negative control.

RESULTSLPS had no significant effect on SCAP proliferation until day 7. RT-PCR assays demonstrated that SCAP expressed SDF-1 mRNA. Different concentrations of LPS significantly promoted the SDF-1 expression in SCAP after 24 h (F = 12.102, P = 0.002) and 48 h (F = 39.054, P < 0.001) exposure, with relative gene expression ratio (experimental/control) increased to 1.4 ± 0.1, 2.2 ± 0.4, 2.3 ± 0.5 in 24 h group and 2.1 ± 0.4, 3.4 ± 0.3, 3.8 ± 0.5 in 48 h group.

CONCLUSIONSIsolated SCAP in cultures have the expression of SDF-1 mRNA. LPS can significantly promote the expression of SDF-1 in SCAP.

Cell Differentiation ; Cell Proliferation ; drug effects ; Chemokine CXCL12 ; genetics ; metabolism ; Dental Papilla ; cytology ; Gene Expression ; Humans ; Lipopolysaccharides ; pharmacology ; Stem Cells ; drug effects ; metabolism

Cell Movement ; drug effects ; Chemokine CXCL12 ; genetics ; metabolism ; pharmacology ; physiology ; Humans ; Neoplasm Invasiveness ; physiopathology ; Neoplasm Metastasis ; physiopathology ; Neoplasms ; metabolism ; Neovascularization, Pathologic ; physiopathology

OBJECTIVETo explore the role of stromal cell-derived factor (SDF-1) and its specific receptor CXCR4 in the G-CSF-induced hematopoietic stem/progenitor cells (HSPCs) mobilization in human healthy donor.

METHODSThe changes of SDF-1/CXCR4 in bone marrow (BM) and peripheral blood (PB) of healthy donors during G-CSF-induced mobilization were detected by enzyme-linked immunosorbent assay (ELISA), immunohistological staining and flow cytometry. SDF-1 neutralizing antibody wes injected into BALB/c mice to further test its effect on mobilization.

RESULTSSDF-1 concentration in mobilized BM (mBM), steady state BM (ssBM) and PB were(7.23 +/- 0.66) microg/L, (5.43 +/- 0.35) microg/L and (5.42 +/- 0.52) microg/L, respectively. SDF-1 protein levels were decreased in the BM (P < 0.05) after 5-day G-CSF injection, and its concentration gradient between BM and PB disappeared (P > 0.05). Significant up-regulation of CXCR4 expression was observed on mBM CD34 cells in healthy donors. The rate of CXCR4 expression on CD34 cells in ssBM, mBM and mobilized PB were (40.98 +/- 21.56)%, (65.80 +/- 24.68)% and (27.54 +/- 26.03)%, respectively. Comparing with that in ssBM and mBM, CXCR4 expression on mobilized PB CD34+ cells were significantly decreased (P < 0.05). Inhibition of SDF-1 signal by blocking monoclonal antibodies significantly reduced G-CSF-induced mobilization in BALB/c mice. This resulted in significant decrease of white blood cell count and progenitors mobilized into peripheral circulation.

CONCLUSIONG-CSF induces HSPCs mobilization by decreasing bone marrow SDF-1 and down-regulating CXCR4 expression on HSPCs.

Animals ; Chemokine CXCL12 ; metabolism ; physiology ; Granulocyte Colony-Stimulating Factor ; pharmacology ; Hematopoietic Stem Cell Mobilization ; methods ; Humans ; Mice ; Mice, Inbred BALB C ; Receptors, CXCR4 ; metabolism ; physiology

OBJECTIVETo explore the effects of advanced oxidation protein products (AOPP) on expressions of stromal cell-derived factor-1alpha (SDF-1alpha) in ECV304 cells and the signal pathway that mediated the effects.

METHODSAOPP-BSA was made from bovine serum albumin (BSA) and sodium hypochlorite. After treated with AOPP-BSA of different concentrations (50, 100, 200 micromol/L), the expressions of SDF-1alpha mRNA in ECV304 cells were measured by reverse transcription-polymerase chain reaction (RT-PCR) and the expressions of SDF-1alpha protein and the levels of phosphorylated extracellular signal-regulated kinase (ERK) in ECV304 cells were analyzed by Western blot. In inhibition test, U0126, the special inhibitor of ERK of different concentrations (0.1, 1, 10 rmol/L) were added into ECV304 cells culture media for 1 hour, then the cells were treated with AOPP-BSA for 24 hours, at last the protein levels in supernatant were detected by enzyme-linked immunosorbent assay (ELISA).

RESULTSAOPP-BSA obviously promoted the expressions of SDF-1alpha mRNA and increased the levels of SDF-1beta protein of ECV304 cells in dose-dependent manner (all P < 0.01), after 15 minutes treated with 200 micromol/L AOPP-BSA, the levels of phosphorylated ERK of ECV304 cells increased significantly (P < 0.01). When the ERK pathway was blocked by U0126, the promoting effects of AOPP-BSA on expressions of SDF-la protein in ECV304 cells were significantly inhibited in dose-dependent manner (P < 0.05).

CONCLUSIONAOPP induced the expression of SDF-la of ECV304 cells, ERK signal pathway is an important pathway that mediated the effects.

Advanced Oxidation Protein Products ; pharmacology ; Cell Line ; Chemokine CXCL12 ; metabolism ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Humans ; MAP Kinase Signaling System ; Oxidative Stress ; Phosphorylation ; RNA, Messenger ; genetics