OBJECTIVETo investigate the effect of oleic acid on the proliferation and secretion of pro-inflammatory mediators of human normal fibroblasts and scar fibroblasts.

METHODSHuman normal fibroblasts and scar fibroblasts were cultured in vitro and respectively divided into seven groups according to the random number table, with 8 samples in each group. Cells in blank control (BC) group were routinely cultured without addition of other agents. Cells in ethanol-control (EC) group were cultured with medium with the addition of 2% absolute ethanol. Cells in oleic acid groups were cultured with the addition of oleic acid in concentration of 0.25, 0.50, 1.00, 2.00, or 4.00 mmol/L in 2% absolute ethanol. The growth of cells in each group was observed with trypan blue staining on post culture day (PCD) 1-5. On PCD 2, structure of cells in BC, EC, and 1.00 mmol/L oleic acid groups was observed under inverted phase contrast microscope and transmission electron microscope; cell cycle of BC, EC, and 1.00 mmol/L oleic acid groups was measured by flow cytometer; cell proliferation activity in each group was measured by MTT assay; the level of NO in supernatant was assayed by Griess assay; the levels of TNF-α, IL-1β, IL-6, and IL-8 in supernatants in each group were determined by enzyme-linked immunosorbent assay. Data were processed with multifactor and repeated measurement design analysis of variance.

RESULTS(1) There was no significant difference in each index of normal fibroblasts and scar fibroblasts between BC group and EC group. (2) The numbers of normal fibroblasts and scar fibroblasts in 2.00 and 4.00 mmol/L oleic acid groups were significantly lower than those in corresponding BC and EC groups on PCD 2-5 (with F values respectively 13.773 and 11.344, P values all below 0.01). (3) On PCD 2, the numbers of normal fibroblasts and scar fibroblasts in 1.00 mmol/L oleic acid groups decreased, and the cells were aggregating, rounding, and easy to drop off. Cellular membrane disruption, vacuolar degeneration of mitochondrion, pyknosis, and lipid droplets were observed. (4) The percentages of G0/G1 and G2/M phases of normal fibroblasts in 1.00 mmol/L oleic acid group [(93.56 ± 9.98)%, (2.01 ± 0.75)%] were significantly higher than those in BC group [(84.23 ± 10.96)%, (0.37 ± 0.16)%, with F values respectively 3.026, 34.751, P < 0.05 or P < 0.01], while the percentage of normal fibroblasts in S phase [(4.42 ± 0.87)%] was markedly lower than that in BC group [(16.06 ± 1.74)%, F = 136.120, P < 0.01]. The percentages of scar fibroblasts of G0/G1 and G2/M phases in 1.00 mmol/L oleic acid group [(93.86 ± 13.90)%, (1.89 ± 0.66)%] were significantly higher than those in BC group [(83.88 ± 10.42)%, (0.41 ± 0.17)%, with F values respectively 3.529, 32.710, P < 0.05 or P < 0.01], and the percentage of scar fibroblasts in S phase [(3.87 ± 0.63)%] was markedly lower than that in BC group [(15.89 ± 2.02)%, F = 116.508, P < 0.01]. (5) The proliferation rates of normal fibroblasts and scar fibroblasts in 0.50-4.00 mmol/L oleic acid groups were significantly lower than those in corresponding BC and EC groups (with F values respectively 215.945 and 194.555, P < 0.05 or P < 0.01). (6) The content of NO in supernatant of normal fibroblasts in all oleic acid groups was obviously higher than that in BC and EC groups (F = 30.240, P < 0.05 or P < 0.01). The contents of NO in supernatants of scar fibroblasts in 1.00-4.00 mmol/L oleic acid groups were significantly higher than that in BC and EC groups (F = 12.495, P < 0.01). The contents of TNF-α and IL-6 in supernatants of normal fibroblasts and scar fibroblasts in 2.00 and 4.00 mmol/L oleic acid groups were obviously higher than those in corresponding BC and EC groups (with F(TNF-α) values respectively 6.911, 3.818, F(IL-6) values respectively 16.939, 11.600,P < 0.05 or P < 0.01). The contents of IL-1β in supernatants of normal fibroblasts and scar fibroblasts in groups of every concentration of oleic acid were significantly higher than those in corresponding BC and EC groups (with F values respectively 25.117, 9.137, P values all below 0.01). The contents of IL-8 in supernatants of normal fibroblasts in 1.00-4.00 mmol/L oleic acid groups were markedly higher than those in BC and EC groups (F = 2.717, P < 0.05 or P < 0.01). The contents of IL-8 in supernatants of scar fibroblasts in 2.00 and 4.00 mmol/L oleic acid groups were significantly higher than those in BC and EC groups (F = 3.338, P < 0.05). There was no statistically significant difference in above indexes between normal fibroblasts and scar fibroblasts in the same concentration of oleic acid group (with F values from 0.120 to 3.766, P values all above 0.05).

CONCLUSIONSAlthough oleic acid in high concentration inhibits the proliferation of scar fibroblasts, it also inhibits the proliferation of normal fibroblasts. Oleic acid in high concentration can cause excessive and continued inflammatory reaction by promoting the secretion of pro-inflammatory mediators of normal fibroblasts and scar fibroblasts, and they are detrimental to wound healing.

Cell Proliferation ; drug effects ; Cells, Cultured ; Cicatrix ; metabolism ; Fibroblasts ; cytology ; drug effects ; secretion ; Humans ; Inflammation Mediators ; metabolism ; Oleic Acid ; pharmacology

The resistance effect on Newcastle disease virus (NDV) and Infectious Bursal Disease Virus(IBDV) in vitro of a new antimicrobial substance (AS), which produced by a Bacillus subtilis strain named B. subtilis fmbJ. Results showed that the TD50 and TD0 value of this AS on Chicken Embryo Fibroblasts cell (CEF) were 128.95mg/L and 25.79mg/L, respectively. This AS could strongly inhibit the cytopathic effects of cell induced by NDV as well as IBDV, and increase the survival rate of cell remarkably. This AS could inhibit the function of NDV and IBDV, and it could defend against the infection and inhibit multiplication of NDV and IBDV, and the effect was the same as the antiviral medicine Ribavirin. It had lower toxicity to CEF cell, therefore we would study it further that it was as antiviral medicine.
Animals ; Antiviral Agents ; metabolism ; toxicity ; Bacillus subtilis ; metabolism ; Chick Embryo ; cytology ; Fibroblasts ; cytology ; drug effects ; Infectious bursal disease virus ; drug effects ; Newcastle disease virus ; drug effects

OBJECTIVETo observe the effects of Angelica dahurica extracts on the biological characteristics of human dermal fibroblasts in vitro and to preliminary explore its possible therapeutic mechanism for wound healing.

METHODSThe optimal concentration of Angelica dahurica extracts was identified by analysing of proliferation activity of human normal fibroblasts (Fb) that treated with different concentration of Angelica dahurica extracts through thiazole blue (MTT) colorimetric assay. Cell cycle, collagen I and collagen III mRNA levels of the optimal Angelica dahurica extracts treated Fb were detected by flow cytometry (FCM) and real-time PCR techniques.

RESULTSAt concentrations of 5 × 10(-4) to 5 × 10(-2) g/L, the Angelica dahurica extracts significantly enhanced the proliferation of Fb. The most significant concentration was 5 × 10(-3) g/L (t = 5.79, P < 0.01), at which an increased percentage of G1 to S and S to G2 phase cells (t = 11.2, 5.69, 2.44, P < 0.05) as well as an increased level of collagen I (1.61 ± 0.26 vs. 1.00 ± 0.16) and collagen III mRNA (3.36 ± 0.40 vs. 1.00 ± 0.14) were obtained compared to the control group (t = 6.69, 7.64, P < 0.01).

CONCLUSIONSAngelica dahurica extracts can notably promote the proliferation of Fb and accelerating the cell cycle of Fb as well as up-regulating the expression of collagen I and collagen III, which may enhance the process of wound healing.

Angelica ; chemistry ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Collagen ; metabolism ; Dermis ; cytology ; Fibroblasts ; cytology ; drug effects ; metabolism ; Humans ; Plant Extracts ; pharmacology

OBJECTIVETo study the effect of insulin in different concentrations on secretion function of growth factors of adipose-derived stem cells (ADSCs).

METHODSADSCs were isolated from human abdominal adipose tissue and cultured. The immunophenotype and adipose induced-differentiation were identified, and the third generation cells were collected. The collected cells were assigned to 1 x 10(-8), 1 x 10(-7), 1 x 10(-6) mol/L insulin groups according to the concentration of added insulin. When cells grew into 70% confluence in conventional medium, ADSCs were cultured further in serum-free DMEM containing insulin in different concentrations for 3 days. ADSCs cultured in medium without insulin were used as control group. Secretion amount of vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) of ADSCs were determined by enzyme-linked immunosorbent assay. The effects of the supernatant fluid of ADSCs' nutrient solution on the proliferation and collagen synthesis of the cultured fibroblast were detected by MTT chromatometry and hydroxyproline chromatometry.

RESULTSThe secretion amounts of VEGF and HGF of ADSCs in 1 x 10(-8) and 1 x 10(-7) mol/L insulin groups [(471 +/- 41, 762 +/- 66 ng/L), (643 +/- 64, 930 +/- 67 ng/L), respectively] were significantly higher as compared with those in control group (286 +/- 47, 577 +/- 84 ng/L) (P < 0.05 or P < 0.01). No change occurred in the secretion amount of VEGF and HGF of ADSCs in 1 x l0(-6) mol/L insulin group (P > 0.05). The supernatant fluid of ADSCs' nutrient medium of 1 x 10(-8), 1 x 10(-7) mol/L insulin groups showed obvious stimulative effect on the proliferation and collagen synthesis of fibroblasts, and it was most obvious in the 1 x 10(-7) mol/L group (P < 0.05 or P < 0.01).

CONCLUSIONSInsulin in the concentrations of 1 x 10(-8) and 1 x 10(-7) mol/L can notably promote ADSCs' function of secreting VEGF and HGF.

Adipocytes ; cytology ; drug effects ; secretion ; Cells, Cultured ; Fibroblasts ; cytology ; Hepatocyte Growth Factor ; metabolism ; Humans ; Insulin ; pharmacology ; Stem Cells ; cytology ; drug effects ; secretion ; Vascular Endothelial Growth Factor A ; metabolism

Cells, Cultured ; Fibroblasts ; cytology ; drug effects ; metabolism ; Humans ; Lung ; cytology ; metabolism ; Macrophages, Alveolar ; metabolism ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Silicon Dioxide ; toxicity ; Silicosis ; metabolism

OBJECTIVE: To observe the effects of xipayi mouth rinse on the DNA synthesis and change of cell cycles of human gingival fibroblast (HGF) induced by lipopolysaccharide(LPS). METHODS: HGF was stimulated with LPS at 25 mg/L. Flow cytometry was used to examine the effect of xipayi mouth rinse at 25 mg/L on the DNA synthesis and change of HGF cell cycles. RESULTS: The percentage of HGF in G( 1) phase increased after the cells were induced by LPS, while the percentage of HGF in S phase decreased. Xipayi mouth rinse could ameliorate this phenomenon. CONCLUSION Xipayi mouth rinse can significantly ameliorate the inhibitory effect of LPS on the proliferation of HGF, suggesting the anti-inflammatory effect of xipayi mouth rinse in the treatment and prevention of periodontal diseases.
Cell Cycle ; drug effects ; DNA ; biosynthesis ; Drugs, Chinese Herbal ; pharmacology ; Fibroblasts ; cytology ; metabolism ; Gingiva ; cytology ; Humans ; Lipopolysaccharides ; Mouthwashes ; pharmacology

OBJECTIVETo investigate the effects of testosterone on the proliferation of penile corpus cavernosal cells in male SD rats.

METHODSSmooth muscle cells (SMCs) and fibroblasts collected from the corpus cavernosal tissues of male SD rats were cultured by the enzymatic dispersion method and detected for the expression of the androgen receptor (AR) by immunohistochemistry. The effects of testosterone on the SMCs and fibroblasts were observed by methyl thiazolyl tetrazolium (MTT) assay in different concentration groups (10(-8) mol/L, 10(-7) mol/L, 10(-6) mol/L, 10(-5) mol/L, 10(-4) mol/L and 10(-3) mol/L) in comparison with the control.

RESULTSThe AR expression was found in the penile corpus cavernosal tissues. MTT assay showed that, at the concentration of 10(-5) mol/L, testosterone induced the proliferation of SMCs (68100 +/- 2200) and fibroblasts (70200 +/- 1300), with significant differences from the control ( P < 0.05), while at 10(-4) mol/L, it inhibited their proliferation (55000 +/- 1400 and 59100 +/- 1500, respectively), (P < 0.01). No significant effects were noted in the other concentration groups.

CONCLUSIONAR exists in the penile corpus cavernosal tissues of male rats. Testosterone modulates the proliferation of corpus cavernosum tissue cells through AR, and different concentrations of testosterone may be positively or negatively correlated with the proliferation of SMCs and fibroblasts.

Animals ; Cell Proliferation ; drug effects ; Cells, Cultured ; Dose-Response Relationship, Drug ; Fibroblasts ; cytology ; drug effects ; metabolism ; Immunohistochemistry ; Male ; Myocytes, Smooth Muscle ; cytology ; drug effects ; metabolism ; Penis ; cytology ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, Androgen ; metabolism ; Testosterone ; pharmacology

OBJECTIVETo investigate the effects of transforming growth factor-β1 (TGF-β1) on growth controlling and the expression of connective tissue growth factor mRNA(CTGF mRNA) in urethra epithelium cells and fibroblasts cultured in vitro.

METHODSUrethra epithelial cells and fibroblasts were cultured in vitro and identified. The fourth generation cells were divided into control group (cultured by cell medium without TGF-β1) and experimental groups(cultured by cell medium containing TGF-β1 1, 2, 4 and 8 µg/L), the vital force of cells were examined by MTT and cell counting, the expression of CTGF mRNA were examined by RT-PCR after 24 hours.

RESULTSThe optical density and cell count decreased in experimental groups of urethra epithelium cells and increased in experimental groups of fibroblasts with the concentration of TGF-β1 being heightened compared with the control group (P < 0.05). The expression of CTGF mRNA increased with the heightening concentration of TGF-β1 in all experimental groups of urethra epithelium cells and fibroblasts by RT-PCR (P < 0.05).

CONCLUSIONSTGF-β1 can inhibit the growth of urethra epithelium cells and promote the growth of fibroblasts in vitro, it can induce the expression of CTGF mRNA in two cells above-mentioned.

Animals ; Cell Survival ; drug effects ; Cells, Cultured ; Connective Tissue Growth Factor ; genetics ; metabolism ; Epithelial Cells ; cytology ; drug effects ; metabolism ; Fibroblasts ; cytology ; drug effects ; metabolism ; Male ; Mucous Membrane ; cytology ; RNA, Messenger ; genetics ; Rabbits ; Transforming Growth Factor beta1 ; pharmacology ; Urethra ; cytology

OBJECTIVETo study the cellular and molecular mechanism of gasoline-induced adverse effects on skin, particularly on keratinocyte and fibroblast in vitro.

METHODSThe primary cell culture of keratinocyte and fibroblast were treated with 0, 0.001%, 0.01%, 0.1% and 1.0% gasoline, respectively. (3)H-thymidine ((3)H-TdR), (3)H-leucine ((3)H-Leu), (3)H-proline ((3)H-Pro) and (14)C-linoleic acid incorporation tests were applied to elucidate their capacity of synthesizing DNA, protein and sebum.

RESULTSThe incorporation of (3)H-TdR in keratinocyte and (3)H-TdR and (3)H-Pro in fibroblast inhibited significantly after exposure to 0.01% gasoline (P < 0.05), with inhibition rates 68.5%, 45.1% and 40.6% for (3)H-TdR in keratinocyte, and (3)H-TdR and (3)H-Pro in fibroblast, respectively. Significant depression in incorporation of (3)H-Leu and (14)C-linoleic acid in keratinocyte were found even in the group treated with 0.001% gasoline (P < 0.05), with inhibition rates of 20.2% and 41.2%, respectively.

CONCLUSIONSSolvent gasoline has certain toxic effect on keratinocyte and fibroblast, intervening their normal metabolic and physiological process and affecting their ability of synthesizing DNA, protein and sebum, and their physiological functions, which could be one of the mechanisms causing skin damage by gasoline. The results also indicated that keratinocyte was more susceptible to gasoline than fibroblast.

Animals ; Cells, Cultured ; DNA ; biosynthesis ; genetics ; Dose-Response Relationship, Drug ; Fibroblasts ; cytology ; drug effects ; metabolism ; Gasoline ; toxicity ; Keratinocytes ; cytology ; drug effects ; metabolism ; Proteins ; drug effects ; metabolism ; Rats ; Rats, Sprague-Dawley ; Sebum ; drug effects ; metabolism

OBJECTIVETo investigate the origin of myofibroblasts in oral submucous fibrosis.

METHODSThe oral keratinocytes and fibroblasts were isolated and cultured. The expression of the alpha-smooth muscle actin in the fibroblasts was examined by immunohistochemistry and reverse transcriptase polymerase chain reaction (RT-PCR).

RESULTSNo difference was found in expression of alpha-smooth muscle actin between the fibroblasts that were directly stimulated by arecoline and the control. The expression of alpha-smooth muscle actin in the keratinocyte and fibroblast-cocultured group was higher than in the control group, and higher in fibroblasts cocultured with keratinocytes preprocessed by arecoline than in fibroblasts cocultured with keratinocytes without preprocessed by arecoline.

CONCLUSIONSThe differentiation of myofibroblasts from fibroblasts in oral submucous fibrosis might be induced by the interaction of arecoline and keratinocyte.

Actins ; metabolism ; Arecoline ; pharmacology ; Cell Differentiation ; drug effects ; Cells, Cultured ; Coculture Techniques ; Fibroblasts ; cytology ; metabolism ; Humans ; Keratinocytes ; cytology ; Mouth Mucosa ; cytology ; Oral Submucous Fibrosis ; metabolism ; pathology