OBJECTIVETo observe the effects of complement fragment C3f on expression and secretion of collagen I, III and transforming growth factor( TGF)-beta1 in human embryonic lung fibroblast (MRC-5) cells.

METHODSMRC-5 cells were cultured with C3f (the synthetic 17 peptides fragments of complement C3). The extracellular and intracellular expression levels of type I, III collagens and TGF-beta1 in MRC-5 cultures were detected by ELISA and immunohistochemistry, respectively.

RESULTSThe expression levels of type I, III collagen and TGF-beta1 in the supernatant of MRC-5 cultures decreased significantly with the concentrations of C3f as compared with controls (P < 0.05). Also the expression level of TGF-beta1 in MRC-5 cytoplasm reduced significantly as compared with controls (P < 0.05).

CONCLUSIONThe results of present in vitro study showed that the complement fragment C3f could reduce the formation of TGF-beta1 and type I, III collagens in MRC-5 cells, and inhibit the lung tissue fibrosis.

Cell Line ; Collagen Type I ; metabolism ; Collagen Type III ; metabolism ; Complement C3b ; pharmacology ; Fibroblasts ; drug effects ; metabolism ; Humans ; Lung ; cytology ; drug effects ; embryology ; Transforming Growth Factor beta1 ; metabolism

The collagen I was made with the dialysis method of enzymolysising the pig skin, and the static deposition in vitro of calcium phosphate was comparative studied by X-ray diffraction (XRD) and infrared spectroscopy (FTIR) under the condition of pH7. 4, Ca/P 1.67 and whether adding the collagen I into the system. Then the chemical composition of the sedimentary product and the diversification of the collagen I 's IR and Raman spectra (RS) before and after the mineralization were analyzed. The results showed that,under the physiological pH condition that there was not any collagen I, though Ca/P reached up to 1.67, the sedimentary product was CaHPO4 x 2H2O yet, however, after adding collagen I into the system, the bone-like apatite was deposited, which proved that collagen I indeed had the effects on the inducing of the bone-like apatite's mineralization in vitro; there was obviously mutual coordination action between collagen I and its mineralization product--bone-like apatite, which caused that amide peak I red-shifted, amide peak II and amide peak III decreased significantly or disappeared on the IR of collagen I, which maybe was the mechanism that how collagen I induced the depositing of the bone-like apatite.
Animals ; Apatites ; metabolism ; Collagen Type I ; pharmacology ; Osteogenesis ; drug effects ; Skin ; chemistry ; Swine

OBJECTIVETo investigate the role of recombinant human transforming growth factor beta3 (rhTGFbeta3) on fibroblast and its possible mechanism.

METHODSNormal skin fibroblast (NSFb) and hypertrophic scar fibroblast (HSFb) were cultured in vitro, and were processed by different concentrations of rhTGFbeta3. NSFb and HSFb in DMEM solution without rhTGFbeta3 were employed as control. The changes in the protein and mRNA expression of type I and III collagen in NSFb and HSFb were observed.

RESULTS(1) The expression of type I and III procollagen in NSFb was evidently different from that of HSFb (2) The synthesis of type I and III procollagen in all test groups was increased obviously after rhTGFbeta3 process (P < 0.001) while the ratio of type I to III procollagen was decreased when compared with that in control group. (3) The effects of rhTGFbeta3 on the biological behavior exhibited an obvious dose- effects relationship. The contents of type I to III procollagen in HSFb were higher than those in NSFb when the dose of rhTGFbeta3 was same.

CONCLUSIONrhTGFbeta3 could effectively promote the synthesis of type I and III procollagen, especially type III procollagen in fibroblasts. This might be beneficial to the accelerate of wound healing and to inhibit or prevent scar formation.

Cicatrix, Hypertrophic ; metabolism ; Collagen Type I ; biosynthesis ; Collagen Type III ; biosynthesis ; Fibroblasts ; drug effects ; metabolism ; Humans ; In Vitro Techniques ; Recombinant Proteins ; pharmacology ; Transforming Growth Factor beta3 ; pharmacology

OBJECTIVETo evaluate the effects of taurine in diet on the expression of type I and III collagen and collagen ratio at different time points in rats lung by image process technology.

METHODSWistar rats were randomly divided into three groups: the saline instilled with a control diet (the saline treated group); silica instilled with a control diet (the silica treated group); and silica instilled with a diet containing 2.5% taurine (the taurine treated group). Animal models were established by the direct tracheal instillation of silica into rat lungs exposed surgically. The taurine concentration of serum was analyzed by means of HPLC. Paraffin embedded lung sections were stained with Sirius red. Polarization microscopy and Image Pro Plus Version 4.5 for windows were used for detecting type I and III collagen.

RESULTSThe concentration of taurine in serum of the taurine treated group was significantly elevated compared to the saline treated and silica treated group (P < 0.05 or P < 0.01). Sirius red polarization microscopy showed that type I and III collagen positive area percentage were elevated in the silica treated rats compared with the saline treated group. On the 7th, 14th, 21st, 28th day after silica instillation type I collagen positive area percentage was increased by 3.84, 3.77, 3.73, 9.83 respectively (P < 0.01), and type III collagen positive area percentage were elevated by a little in the silica treated rats compared with saline treated group. The taurine treatment significantly decreased elevation of silica type I collagen positive area percentage of lung by 2.39, 1.62, 7.13 at the 7th, 21st, 28th day respectively (P < 0.05 or P < 0.01), and type III collagen positive area percentage of lung by 2.62 at the 28th day (P < 0.05) compared with the silica treated group. The ratio of type I to III collagen was increased from the 7th day to 28th day after silica instillation, and reached 1.87 at the 28th day with the maximal ratio in the silica-treated group.

CONCLUSIONTreatment with taurine can effectively attenuate type I and III collagen expression in the rat lung induced by silica particles at different time points in our study.

Animals ; Collagen Type I ; biosynthesis ; Collagen Type III ; biosynthesis ; Female ; Lung ; drug effects ; metabolism ; Male ; Random Allocation ; Rats ; Rats, Wistar ; Silicon Dioxide ; toxicity ; Taurine ; pharmacology

OBJECTIVETo investigate the effects of ethanolic extract from Capparis spinosa (ECS) on the fibroblast proliferation and type I collagen production in normal and progressive systemic sclerosis (PSS).

METHODCellular activity was determined by the MTT method. Apoptosis was detected by flow cytometry analysis of Annexin V-stained cells. The expression levels of type I collagen messenger RNA and protein were analyzed by RT-PCR and western blot analysis.

RESULTECS could significantly inhibit the proliferation of fibroblast and reduced the expression of alpha2 (I) collagen mRNA and type I collagen protein in PSS in a dose-and time-dependent manner. ECS did not affect the proliferation of fibroblast and expression of type I collagen mRNA and protein in normal human. ECS could counteract the harmful effects on fibroblast by H2O2.

CONCLUSIONECS can effectively inhibit the fibroblast proliferation and type I collagen production in PSS.

Capparis ; chemistry ; Cell Proliferation ; drug effects ; Cells, Cultured ; Collagen Type I ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Fibroblasts ; cytology ; drug effects ; Humans ; Scleroderma, Diffuse ; drug therapy ; metabolism ; pathology

OBJECTIVETo seek an effective drug for treatment of human hyperplastic scar through studying the effects of curcumin on fibroblast growth and collagen synthesis.

METHODSFibroblasts derived from scar tissue and from normal epidermal tissue were isolated and cultured separately with tissue-block method, their morphology were observed under invert phase contrast microscope, their growth curve was drawn respectively to determine the speed of growth. Then, fibroblasts from scar were stimulated with curcumin in different concentrations (0, 12.5, 25, 50 and 100 micromol/L) for detecting the inhibitory effect of curcumin on growth of fibroblasts using MTT methods and that on activity of procollagen alpha-1 gene transcription in fibroblast was detected by RT-PCR.

RESULTSThe cell growth curve showed that double-multiplying time was 5 days in fibroblasts from scar and 4 days in those from normal dermis, showing significant difference between them (P < 0.05). MTT showed that curcumin in 12.5 micromol/L showed a cell proliferation enhancing trend, and its absorbance value was significantly higher than that in the normal group, but the effect turned to inhibition when concentration increased to over 25-100 micromol/L, and became significant inhibition at concentration of 50 and 100 micromol/L. Besides, curcumin also showed markedly inhibition on collagen type I synthesis in fibroblasts (P < 0.01).

CONCLUSIONHigh concentration curcumin can inhibit effectively the fibroblast proliferation and collagen I synthesis in hyperplastic scar, therefore, may has therapeutic effect on the disease in human being.

Cell Proliferation ; drug effects ; Cells, Cultured ; Cicatrix, Hypertrophic ; metabolism ; pathology ; Collagen Type I ; biosynthesis ; Curcumin ; pharmacology ; Fibroblasts ; drug effects ; metabolism ; pathology ; Humans

To provide a support to the clinical application of alendronate (Alen) on cytology, we studied the effects of Alen on the function of osteoblasts. In this experiment, we observed the influence of MG63 cell line co-incubation with Alen at concentrations of 1 x 10(-9) mol/L, 1 x 10(-7) mol/L or 1 x 10(-5) mol/L on the osteoblastic function (proliferation, cell morphology, alkali phosphatase (ALP) activity, expression of type I collagen and effect of calcium deposition). The proliferation, cell morphology, ALP activity and type I collagen synthesis of MG63 were not affected by Alen at concentration of 1 x 10(-9) mol/L and 1 x 10(-7) mol/L, but the ALP activity as well as type I collagen production were promoted at higher concentration (1 x 10(-5) mol/L). The calcium deposition of MG63 could be increased at the lower concentration (1 x 10(-9) mol/L), while it was inhibited at the higher concentration. In conclusion, Alen at low concentration can promote the mineralization ability of osteoblasts to a certain extent, and this benefits the bone formation.
Alendronate ; pharmacology ; Alkaline Phosphatase ; metabolism ; Bone Density Conservation Agents ; pharmacology ; Cell Line ; Cell Proliferation ; drug effects ; Collagen Type I ; metabolism ; Humans ; Osteoblasts ; cytology ; Osteogenesis ; drug effects

OBJECTIVESTo study the effects of Sanqi qisodium hyaluronate gel on collagen-I and collagen-III expression in the process of rabbits' epidural scar formation after operation.

METHODSNinety-six white rabbits with 6-month-old, half males and half females, weighted from 2 to 2.5 kg, which were randomly divided into normal saline group (A), Sanqi group (B), qisodium hyaluronate group (C) and Sanqi qisodium hyaluronate gel group (D). The laminectomy of rabbits were performed in group A, B, C, D, the duras were surrounded with normal saline, Sanqi liquid, qisodium hyaluronate and Sanqi qisodium hyaluronate gel respectively. Animals of each group were killed at 1, 2, 4, 8 weeks after operation. Use Masson staining for histological observation of collagen, and in situ hybridization staining for the analysis of collagen-I and collagen-III expression.

RESULTSIn the Masson staining, Sanqi qisodium hyaluronate gel group was more regular than the control group in the shape of collagen texture. As to the expression of collagen-I, and Sanqi qisodium hyaluronate gel group was lower than normal saline group, the Sanqi group and qisodium hyaluronate group at 4 weeks after using medicine (P < 0.01); while the Sanqi qisodium hyaluronate gel group was higher than normal saline group, Sanqi group and qisodium hyaluronate group in the collagen-III expression (P < 0.01).

CONCLUSIONSSanqi qisodium hyaluronate gel could improve collagen's arrangement of the rabbit's epidural scar after operation, reduce its rigidity and increase flexibility.

Animals ; Cicatrix ; etiology ; metabolism ; Collagen Type I ; genetics ; metabolism ; Collagen Type III ; genetics ; metabolism ; Epidural Space ; Female ; Gels ; Gene Expression Regulation ; drug effects ; Hyaluronic Acid ; chemistry ; Laminectomy ; adverse effects ; Male ; RNA, Messenger ; genetics ; metabolism ; Rabbits

OBJECTIVETo investigate the effect of salvianolic acid B (SAB) on the proliferation of human embryonic lung fibroblast MRC-5, and the secretion of procollagen I and endogenous transforming growth factor-beta1, (TGF-beta1).

METHODSThe MRC-5 cells were randomly divided into four groups as follows: the control group: cells cultured with DMEM but with no TGF-beta1, or SAB; the TGF-beta1, group: cell cultured with 10 ng/mL TGF-beta1; the SAB1 group: cell cultured with medium with 10 ng/mL TGF-beta1 and 1 pmol/L SAB; the SAB2 group: cell cultured with medium with 10 ng/mL TGF-beta1, and 10 pmol/L SAB. The proliferation of cells was assayed by MTT incorporation. The concentration of amino-terminal propeptide of type I procollagen (PINP), a marker of collagen synthesis, was measured by radioimmunoassay. The endogenous TGF-beta1, levels were measured using ELISA.

RESULTSThe optical density, procollagen I contents, and endogenous TGF-beta1, levels significantly increased when compared with those of the control group (P<0.05). Compared with the TGF-beta1, group, the optical density was obviously lowered, the procollagen I contents and endogenous TGF-beta1, levels significantly decreased in the SAB1 group and the SAB2 group, and better in the SAB2 group, showing statistical difference (P<0.05).

CONCLUSIONSSAB could inhibit the proliferation of MRC-5 cells induced by TGF-beta1 and attenuate the roles of secreting collagen and endogenous TGF-beta1. It had the potential of postponing or delaying the progressive developing of pulmonary fibrosis.

Benzofurans ; pharmacology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Collagen Type I ; metabolism ; Fibroblasts ; cytology ; drug effects ; Humans ; Lung ; cytology ; embryology ; Transforming Growth Factor beta1 ; pharmacology

OBJECTIVETo explore the effects of rosiglitazone (RSG), an agonist of peroxisome proliferators-activated receptor-gamma (PPAR-gamma), on the up-regulation of connective tissue growth factor (CTGF) and the deposition of type I and type III collagens in the pulmonary arteries of rats suffering from fibrosis in lung.

METHODSForty-eight male Sprague-Dawley rats were randomly divided into 4 groups: bleomycin (BLM) plus normal saline (NS) group (n=21), BLM plus RSG group (n=9), NS plus NS group (n=9), and NS plus RSG group (n=9). The rats were received single intratracheal instillation of BLM (5 mg/kg bw) or equal volume of NS as control, and received intra-gastric adminnistration of RSG (3 mg/(kg x day), 14 day) or the same volume of NS as vehicle. In vio, the observation was conducted on day 14 after intratracheal instillation. In vitro, the pulmonary arteries of rats on day 14 after BLM were isolated and incubated with DMEM alone or with RSG (37 degrees C, 5% CO2, for 24 h.

RESULTSIn vivo, the expression and the content of CTGF, the contents of type I and type III collagens, and the ratio of type I collagen and type III collagen were increased in the pulmonary arteries of BLM-instilled rats, compared with those of NS-instilled rats (All P < 0.05). The above abnormal changes were ameliorated by RSG (All P < 0.05). In vitro, RSG blocked the up-regulation of CTGF (P < 0.05), but not the deposition of type I collagen and type III collagen in the pulmonary arteries isolated from the BLM-instilled rats (P > 0.05).

CONCLUSIONThe results suggest that RSG directly blocks the up-regulation of CTGF in pulmonary arteries of rats suffering from fibrosis in lung, and this might be one of the mechanisms underling the ameliorated pulmonary arterial remodeling by RSG.

Animals ; Bleomycin ; Collagen Type I ; metabolism ; Collagen Type III ; metabolism ; Connective Tissue Growth Factor ; metabolism ; Down-Regulation ; drug effects ; Lung ; pathology ; Male ; PPAR gamma ; agonists ; Pulmonary Artery ; metabolism ; Pulmonary Fibrosis ; chemically induced ; metabolism ; physiopathology ; Rats ; Rats, Sprague-Dawley ; Thiazolidinediones ; pharmacology