BACKGROUND: Recent clinial observations have suggested that colchicine, which is in frequent use in gout can affect the conneciive tissue metabolism in skin and other ti.ues. OBJECTIVE: This study suggest that further development of colchiine might provide a novel means of modulating collagen gen expression in patients with fibrotic disease. METHOD: We examed the effect of colchicine on the expres, on of collagen, fibronectin and collagenase gene by skin filroblast culture hy Nort.hern and dot-blot iybridization. Result : The rate of transcription of genomic DNA corresponding o type I collagen and libvonectin was reduced in colchicine-treated cultures but collagenase was not. reduced. Canclusion : The microti.ikile disruptive agent, colchicine, reduced the expression of type I collagen and fibronectin in a dose-(lependent manner. This st.udy suppor that colchicine is one of the promising antifibrogenic drugs curvently being tested as a treatment, of hun an fibrotic disease.
Colchicine* ; Collagen Type I ; Collagen* ; Collagenases ; DNA ; Fibroblasts ; Fibronectins ; Gene Expression* ; Gout ; Humans ; Metabolism ; Skin

It is very difficult to diagnosis osteoarthritis in the early stage, due to the slow development of the disease, no symptoms occures, and no X-ray findings in the early stage, it is difficult to early diagnosis with the traditional diagnostic methods, resulting in the poor treatment outcome, and even some patients develop joint deformity, activity limitation, and must take an operation, it brought great pain and heavy financial burden to patients. How to early diagnosis of articular cartilage injury become difficult now. Some scholars suggest that to those suspected patients, the arthroscopic diagnosis must be taken. Although the small trauma and quick recover, the method of operation has trauma, not only increase the suffering of the patients, but the treatment is very expensive, make the patients finch. A large number of domestic and foreign scholars to study patients with OA to find the ideal fluid biological markers (BM) to reflect articular cartilage metabolism, and revealed disease activity or prognosis. The CTX-II can reflect degradation of the articular cartilage.
Biomarkers ; metabolism ; Collagen Type I ; metabolism ; Humans ; Osteoarthritis ; diagnosis ; metabolism ; Peptides ; metabolism

OBJECTIVETo explore the influence of different thawing temperatures on the morphology and type I collagen metabolism of the human fibroblasts processed at - 10 degrees C in vitro.

METHODSIn vitro cultured human fibroblasts were randomly divided into control, 20 degrees C thawing, and 37 degrees C thawing groups. After being frozen at -10 degrees C, the cells in the latter two groups were thawed at 20 degrees C and 37 degrees C, respectively. The cell proliferation was assessed with MTT method and was expressed by absorption under 570nm (A ), The morphological change of the cells was observed with inverted phase contrast microscope, the change in the intracellular content of collagen was determined with immunohistochemistry, and the extracellular content of collagen was assayed with ELISA.

RESULTSIn 20 degrees C thawing group, the absorbance decreased at first and increased thereafter, and they were obviously lower than that before freezing (0.95 +/- 0.16, P < 0.05 or 0.01). Cell dehydration and shrinking, cytoplasm loss and increased ratio of cytoplasm to nucleus were found in the survived fibroblasts. The cells proliferated actively at 72 and 96 hours after injury, with increased mitotic index and disordered arrangement. Compared with that before freezing (96.4 +/- 2.9) , the extracellular collagen content increased at first, decreased thereafter, and increased again slowly later (P < 0.05), while the intracellular collagen content decreased at first and increased thereafter (P < 0.05). The collagen metabolism in 37 degrees C thawing group was no difference compared with that in control group. Some cells undergone a floating period before adhering to the culture dish walls.

CONCLUSIONCell dehydration after low temperature treatment could protect the cells from damage. Proper thawing temperature could be beneficial to the cell resuscitation. Comparing with slow thawing, rapid thawing could minimize the cell damage.

Cells, Cultured ; Collagen Type I ; metabolism ; Cryopreservation ; Fibroblasts ; cytology ; metabolism ; Freezing ; Humans

To ascertain whether connective tissue growth factor (CTGF) participates in the remodeling of pulmonary artery at the early-stage of bleomycin (BLM)-induced pulmonary fibrosis, mean pulmonary arterial pressure, the expression of type I and type III collagens, and the expression and location of CTGF in pulmonary artery and arteriole were investigated in the present study. Sprague-Dawley rats received instillation of BLM [5 mg/kg body weight, in 0.5 mL of normal saline (NS)] or instillation of the same amount of NS as control. Mean pulmonary arterial pressure was detected via a catheter in the pulmonary artery. Type I and type III collagens were examined with Sirius red staining under polarized light. CTGF expression was investigated by using immunohistochemistry, and was represented as average optical density and percentage of positive area of CTGF. The mean pulmonary arterial pressure was higher in rats on day 14 after BLM instillation [(19.5+/-2.9) mmHg] than that in the control rats [(14.8+/-1.2) mmHg] (P<0.05). The type I and type III collagens were increased both in pulmonary artery and arteriole of rats on day 14 after BLM instillation, compared with those in the control rats (P<0.05, P<0.01, respectively). The ratio of type I/III collagens in pulmonary artery was also higher in BLM-treated rats than that in the control rats (P<0.05). The values of average optical density of positive CTGF staining were increased both in pulmonary artery (0.37+/-0.02) and arteriole (0.40+/-0.03) of rats on day 14 after BLM instillation, compared with those in the control rats (artery, 0.34+/-0.01; arteriole, 0.29+/-0.01) (both P<0.05). The percentages of positive area of CTGF were higher in pulmonary artery (8.40+/-1.13) and arteriole (12.4+/-2.0) of rats on day 14 after BLM instillation than those in the control rats (artery: 1.42+/-0.63; arteriole: 1.16+/-0.34), respectively (both P<0.05). The increased positive CTGF staining areas were mainly located in the endothelium and smooth muscle layer. It is therefore concluded that CTGF expression increases in the endothelium and smooth muscle layer of pulmonary artery and arterioles during high pulmonary arterial pressure and remodeling of pulmonary artery at the early-stage of BLM-induced pulmonary fibrosis, and that the increased CTGF might be one of the mechanisms of maintenance and development of pulmonary hypertension.
Animals ; Bleomycin ; Collagen Type I ; metabolism ; Collagen Type III ; metabolism ; Connective Tissue Growth Factor ; metabolism ; Hypertension, Pulmonary ; Pulmonary Artery ; metabolism ; Pulmonary Fibrosis ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the expression of connective tissue growth factor (CTGF) and its significance in sporadic ascending thoracic aortic aneurysm (AAA), and initially to investigate the mechanisms of pathological remodeling in AAA.

METHODSAAA specimens were taken from 18 patients during elective surgical intervention, and 18 control specimens of ascending aorta were obtained from patients undergoing coronary artery bypass surgery. Specimens were stained with HE and Masson to evaluate the arrangement and aggregation of cells and collagen types I and III; immunohistochemistry staining was performed using antibodies directed against markers of CTGF; real-time PCR analysis was performed to quantify the expression level of CTGF and collagen types I and III.

RESULTSPathological results show degradation of elastin and hyperplasia of collagen fibers as well as disordered arrangement of smooth muscle cells in AAA. When compared with controls, protein levels of CTGF were significantly increased [(44 ± 4)% vs. (33 ± 5)%, P < 0.01]. Similar patterns were shown in mRNA levels of CTGF (P < 0.01). Using real-time PCR method, elevated levels (relative expression ratio of mRNA: 10.54/3.8 and 1.79/1.19, respectively; P < 0.01, both) of collagen types I and III were observed. CTGF expression had a correlation with both collagen fibers and aortic aneurysm diameter (r = 0.784, P < 0.01; r = 0.793, P < 0.01).

CONCLUSIONSThese results indicate increased expression of aortic collagen types I and III as well as CTGF in AAA specimens, which is likely to be responsible for the aortic wall pathological remodeling. The expression of CTGF was positively correlated with the aortic diameter. As a cytokines factor can stimulate collagen synthesis, CTGF may be involved in the pathogenesis and progression of AAA.

Aged ; Aorta ; metabolism ; pathology ; Aortic Aneurysm, Thoracic ; metabolism ; pathology ; Collagen Type I ; metabolism ; Collagen Type III ; metabolism ; Connective Tissue Growth Factor ; metabolism ; Female ; Humans ; Male ; Middle Aged

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

OBJECTIVETo explore the characteristics of LN and type I, III collagen in pulmonary fibrosis induced by uranium ore dust in rats.

METHODS60 adult Wistar rats were divided randomly into two groups, control group (30 rats) and uranium ore dust group (30 rats). Non-exposed intratracheal instillation method was used. Uranium ore dust group was exposed 20 mg/ml uranium ore dust suspension 1ml per rat, meanwhile control group was exposed normal saline 1ml per rat. Post-exposed the 7, 14, 21, 30 and 60 d, 6 rats in each group were killed randomly, lung tissue were collected. The pathological changes in lung tissue were observed by microscope using HE staining, the collagen I and III in lungs were observed by polarizing microscope using Biebrich scarlet staining. The expression of LN protein in lung tissue was observed by immunohistochemistry-SP.

RESULTSDuring lung fibrosis, a large amount of the proliferated I and III collagen in lungs were observed. Post-exposure to uranium ore dust, the characteristics in proliferated collagen in lungs were type I collagen deposited in lung interstitium mainly in the early stage. The area percentage of collagen I and III was increased significantly at 7, 14, 21, 30 and 60d in the experimental group as compared with that in the control group (P < 0.05 or P < 0.01). The over expression of LN in the lung tissue were observed. The expression of LN was distributed in the lung tissue as thickening of the linear or cluster. The integral optical density of LN was increased significantly at 21, 30 and 60 d in the experimental group as compared with that in the control group (P < 0.05 or P < 0.01).

CONCLUSIONSAfter exposure to uranium ore dust, the characteristics in proliferated collagen in lungs are the type of I collagen deposited in lung interstitium mainly in the early stage, while the type of III collagen increase significantly at the later period. The overexpression of LN exists in the process of pulmonary fibrosis. It suggests that LN has a role effect in the process of pulmonary fibrosis.

Animals ; Collagen Type I ; metabolism ; Collagen Type III ; metabolism ; Dust ; Female ; Laminin ; metabolism ; Male ; Pulmonary Fibrosis ; chemically induced ; metabolism ; pathology ; Rats ; Rats, Wistar ; Uranium ; adverse effects

OBJECTIVES: To investigate the patterns of biochemical bone markers, such as urinary deoxypyridinoline (DPD), N-telopeptide of type I collagen (NTX), and serum osteocalcin (OC), bone-specific alkaline pbosphatase (BSAP) in postmenopansal women with hormone replacement therapy (HRT). Materials and METHOD: From July 1997 to January 1998, total 239 postmenopausal women were emolled in the present study, and 198 healthy premenopausal women with regular menstruation were served as control. The postmenopausal women were pouped into the HRT group and the non-HRT group. The women in the HRT poup have received estrogen with continuous or cyclic progestin therapy far more tban 6 months. The biochemical bone markers of all women were assayed. Results were analysed with Students t-test. RESULTS: The urinary DPD of the non-HRT group was sigaificantly higher than both the HRT poup and the premenopausal group(5.51 +/- 2.47 vs. 3.36 +/- 1.02 and 4.01 +/- 3.86 nM/mM, p < 0.05, repectively). The urinary NTX of the non-HRT group was also higher in comparison to the HRT group and the premenopausal group(48.71 +/- 11.54 vs. 33.70 +/- 17.43 and 33.70 +/- 17.43 nM BCE/mmol, p < 0.05, repectively). However, there were no significant differences in the concentrations of serum BSAP and OC among the three poups. CONCLUSION: The urinary DPD and NTX were more sensitive indicators of bone metabolism tban serum BSAP and OC in postmenopausal women undergoing HRT.
Collagen Type I ; Estrogens ; Female ; Hormone Replacement Therapy* ; Humans ; Menstruation ; Metabolism ; Osteocalcin

PURPOSE: The object of this study is to evaluate bone metabolism in healthy adolescents according to the progression of puberty. METHODS: Forty boys(13.9+/-1.7 years) and 42 girls(12.1+/-1.6 years) were classified by Tanner stage (TS) and bone age. Serum levels of osteocalcin(OC) and bone specific alkaline phosphatase(BALP) were measured as bone formation markers. Serum level of C-terminal telopeptide of type I collagen (ICTP) and urinary N-terminal telopeptide of type I collagen(NTx) concentrations adjusted by creatinine concentrations were measured as bone resorption markers. Serum or urine levels of bone turnover markers in each pubertal development group and bone age group were analysed. RESULTS: In boys, BALP and OC levels increased to peak levels significantly(P<0.05), and decreased significantly(P<0.05) from the peak levels to the levels at TS 5. ICTP and NTx levels seemed to increase to peak levels and to decrease from the peak levels to the levels at TS 5. But there were no significant differences except decreasing NTx levels. All showed peak levels between 13 and 15 years of bone age. In comparison with each TS group, BALP and OC levels were significantly different(P<0.05) between each TS group, but ICTP and NTx were not. In girls, the levels of all bone markers seemed to increase to peak levels without significance, and then decrease significantly(P< 0.05). All showed peak levels between 11 and 13 years of bone age. All except ICTP level were significantly different between each TS group(P<0.05). CONCLUSION: The bone metabolism seems to increase as progression of puberty, and to decrease during late puberty. Bone formation markers levels change more actively, rather than bone resorption markers levels during puberty. And the increment of bone formation in early puberty is more significant in boys rather than in girls.
Adolescent ; Bone Resorption ; Collagen Type I ; Creatinine ; Female ; Humans ; Metabolism* ; Osteogenesis ; Puberty*

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