OBJECTIVETo observe the in vitro effect of compound 861 (Cpd 861) on tissue inhibitor of metalloprotenase 1 (TIMP1) mRNA levels of HSC-T6 cell.

METHODSHSC-T6 cells were exposed in different concentrations of Cpd 861 (0.25~1.0 mg/ml) for 48 hours. The TIMP1 level was measured by the quantitative reverse-transcription polymerase chain reaction (RT-PCR).

RESULTSThe TIMP1 mRNA levels of HSC-T6 cells at different concentrations of Cpd 861 were lower (2.50 0.71, 0.50 0.01, 0.11 0.03) than those of the normal control (3.78 0.67, P<0.05 or P<0.01).

CONCLUSIONSThe antifibrotic mechanism of Cpd 861 is partly due to its downregulation on TIMP1 mRNA levels of HSC-T6 cells.

Animals ; Down-Regulation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Gene Expression ; drug effects ; RNA, Messenger ; biosynthesis ; drug effects ; Rats ; Rats, Sprague-Dawley ; Tissue Inhibitor of Metalloproteinase-1 ; biosynthesis ; genetics

OBJECTIVETo study the effect of laminarin polysaccharide (LP) on the activity of matrix metalloproteinase of photoaging skins.

METHODKunming SPF mice were prepared with back hair shaved, and randomly divided into the control group, the model group, the LP low does group (LP-L, 1 mg x kg(-1)), the LP high dose group (LP-H, 5 mg x kg(-1)) and the Vit E (100 mg x kg(-1)) group. They were abdominally injected with drugs twice on a daily basis. Except for the control group, all groups were exposed to ultraviolet rays for 1 hour every day, five times on a weekly basis, with accumulated exposure dose of UVB being 21.60 J x cm(-2) and accumulated exposure dose of UVA being 84.02 J x cm(-2). Eight weeks later, exposed back skins were collected to detect thickness of dermis by HE stain, content of hydroxyproline (Hyp) by chemical colorimetry, and serum MMP-1 and TIMP-1 content by ELISA. In addition, matrix metalloproteinase-1 (MMP-1) mRNA and relative content of tissue inhibitor of metalloproteinase-1 (TIMP1) mRNA was analyzed with Real-time PCR.

RESULTCompared with the model group, the LP-H group could significantly increase the thickness of dermis, skin Hyp content and serum TIMP-1 level, and decrease relative content of MMP-1 mRNA in skin and MMP-1 content in serum.

CONCLUSIONLP can regulate the metabolism of collagen photoaging skins by adjusting the activity of matrix metalloproteinase.

Animals ; Female ; Glucans ; Matrix Metalloproteinase 13 ; biosynthesis ; genetics ; metabolism ; Mice ; Plant Extracts ; chemistry ; pharmacology ; Plants, Medicinal ; chemistry ; Polysaccharides ; chemistry ; pharmacology ; RNA, Messenger ; genetics ; metabolism ; Skin Aging ; drug effects ; physiology ; radiation effects ; Tissue Inhibitor of Metalloproteinase-1 ; biosynthesis ; genetics ; metabolism ; Ultraviolet Rays

OBJECTIVESTo study the effects of c9,t11-conjugated linoleic acid (c9,t11-CLA) on invasive ability of human gastric carcinoma cell line (SGC-7901) and to explore its possible mechanism.

METHODSReconstituted basement membrane invasion assay was used to evaluate invasive ability of cancer cells. Expression of TIMP-1, TIMP-2 and nm23-H(1) mRNA was measured by reverse transcription polymerase chain reaction (RT-PCR) in SGC-7901 cells.

RESULTSAt the concentrations of 200 micromol/L, 100 micromol/L and 50 micromol/L, c9,t11-CLA suppressed their reconstituted basement membrane invasion of SGC-7901 by 53.7%, 40.9% and 29.3%, respectively. c9,t11-CLA could induce the expression of TIMP-1, TIMP-2 and nm23-H(1) mRNA in SGC-7901 cells.

CONCLUSIONSThe invasion of SGC-7901 cells could be inhibited by c9,t11-CLA through reconstituted basement membrane. Anti-invasion action of c9,t11-CLA might be associated with induction of expression of TIMP-1, TIMP-2 and nm23-H(1) mRNA in tumor cells.

Adenocarcinoma ; pathology ; Gene Expression ; drug effects ; Humans ; Linoleic Acid ; pharmacology ; therapeutic use ; Monomeric GTP-Binding Proteins ; biosynthesis ; genetics ; NM23 Nucleoside Diphosphate Kinases ; Neoplasm Invasiveness ; prevention & control ; Nucleoside-Diphosphate Kinase ; RNA, Messenger ; biosynthesis ; drug effects ; Stomach Neoplasms ; pathology ; Tissue Inhibitor of Metalloproteinase-1 ; biosynthesis ; genetics ; Tissue Inhibitor of Metalloproteinase-2 ; biosynthesis ; genetics ; Transcription Factors ; biosynthesis ; genetics ; Tumor Cells, Cultured

OBJECTIVETo study the effect of SiO(2) on the expression of matrix metalloproteinase (MMP) and tissue inhibitor of metalloproteinase (TIMP) in human alveolar macrophages (AMs) associated with the pathogenesis of silicotic fibrosis.

METHODSAMs were collected from a silicotic patient by bronchoalveolar lavage, and exposed to SiO(2) (50 microg/ml), and cultured in DMEM without serum for different time (2, 6, 12, 18, 24, 36 h). Immunocytochemical method was used to detect the level of expression of MMP-9 and TIMP-1 in AMs.

RESULTSThe expression of MMP-9 in AMs exposed to silica was up-regulated, and reached the peak at 18 h [average optical density: (0.440 +/- 0.021) vs (0.390 +/- 0.011), P < 0.05]. After that, the expression reduced markedly. However, the expression of TIMP-1 of AMs were not significantly different from the control group [average optical density: (0.175 +/- 0.019) vs (0.162 +/- 0.044), P > 0.05].

CONCLUSIONSiO(2) could induce up-expression of MMP-9 in AMs. Degradation of basement membrane by MMP-9 produced by AMs at early stage of lung injury may associate with the immigration of various cells including lung fibroblasts into the injured region.

Bronchoalveolar Lavage Fluid ; cytology ; Cells, Cultured ; Humans ; Immunohistochemistry ; Macrophages, Alveolar ; drug effects ; metabolism ; Male ; Matrix Metalloproteinase 9 ; biosynthesis ; Middle Aged ; Silicon Dioxide ; pharmacology ; Silicosis ; pathology ; Tissue Inhibitor of Metalloproteinase-1 ; biosynthesis

Aldosterone ; pharmacology ; Animals ; Cells, Cultured ; Collagen Type I ; biosynthesis ; Collagen Type III ; biosynthesis ; Dose-Response Relationship, Drug ; Liver ; cytology ; drug effects ; metabolism ; Liver Cirrhosis ; etiology ; RNA, Messenger ; analysis ; Rats ; Tissue Inhibitor of Metalloproteinase-1 ; genetics

OBJECTIVETo study the effects of Yiqi Huayu Bushen Recipe (YHBR) and its disassembled recipes on mRNA expressions of collagen I, III, X, matrix metalloproteinase (MMPs) and tissue inhibitor of metalloproteinase (TIMP) in extracellular matrix of cervical disc in model rats of cervical vertebral disc degeneration.

METHODSThe mRNA expressions of collagens, MMP-13 and TIMP-1 were detected by RT-PCR. The strips were scanned by gel imaging system scanner, and the optical density was autocalculated by computer.

RESULTSCompared with those of the sham-operative group, the mRNA expressions of collagen I , Ill and X and MMP-13 of the model rats increased markedly (P < 0.01), which were lowered by YHBR and its disassembled recipes (P < 0.01 or P < 0.05), and the levels after YHBR treatment were significantly different to those after Western medicine treatment. However, no remarkable change was found in TIMP-1 mRNA expression in the model rats (P > 0.05).

CONCLUSIONIn the degenerated intervertebral disc the mRNA expressions of collagen I , III, X and MMP-13 increased, TIMP-1 mRNA expression decreased and the proportion of MMPs/TIMP was in imbalance. YHBR and its disassembled recipes could postpone the degeneration of intervertebral disc through regulating mRNA expressions of collagens and their correlated metabolic enzymes.

Animals ; Cervical Vertebrae ; Collagen ; biosynthesis ; genetics ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Extracellular Matrix ; drug effects ; metabolism ; Intervertebral Disc ; drug effects ; metabolism ; pathology ; Intervertebral Disc Displacement ; drug therapy ; genetics ; metabolism ; Male ; Matrix Metalloproteinase 13 ; biosynthesis ; genetics ; Phytotherapy ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Tissue Inhibitor of Metalloproteinase-1 ; biosynthesis ; genetics

OBJECTIVETo observe the influence of intra-articular injection of sodium hyaluronate (HA) on the mRNA expressions of matrix metalloproteinase-1,-3 (MMP-1,-3) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in cartilage and synovium of traumatic osteoarthritis (OA).

METHODSSixteen white rabbits underwent unilateral anterior cruciate ligament transection (ACLT) were divided into 2 groups randomly 5 weeks after transection. The experimental group rabbits received 0.3 ml of 1% HA by intra-articular injection once a week. Animals in the control group were treated under the same conditions using physiological saline. Ten weeks following surgery, cartilage and synovium were harvested. The mRNA expressions of MMP-1, MMP-3 and TIMP-1 were analyzed using reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSIn synovium, the mRNA expression of MMP-3 was suppressed in the HA injection group. HA treatment had no effect on the MMP-3 expression in cartilage. No significant difference of MMP-1 and TIMP-1 expressions in cartilage and synovium was found between the HA injection group and the control group.

CONCLUSIONSOne of the mechanisms of the therapeutic effect of HA may be the inhibition of expression of MMP-3 in synovium during early stage of traumatic OA.

Animals ; Anterior Cruciate Ligament ; drug effects ; metabolism ; Cartilage, Articular ; metabolism ; Hyaluronic Acid ; pharmacology ; Matrix Metalloproteinase 3 ; biosynthesis ; drug effects ; genetics ; Osteoarthritis, Knee ; metabolism ; pathology ; RNA, Messenger ; genetics ; Rabbits ; Reverse Transcriptase Polymerase Chain Reaction ; Synovial Membrane ; metabolism ; Tissue Inhibitor of Metalloproteinase-1 ; biosynthesis ; drug effects ; genetics

BACKGROUNDIt is known that ultraviolet irradiation can affect cellular function through a number of signaling pathways. (-)-epigallocatechin-3-gallate (EGCG) is the major effective component in green tea and can offer protection from ultraviolet-induced damage. In this study, we investigated the protective mechanism of EGCG on human dermal fibroblasts damaged by ultraviolet A (UVA) in vitro.

METHODSTranscription factor Jun protein levels were measured by Western blot. Matrix metalloproteinase 1 (MMP-1) and tissue inhibitor of metalloproteinase-1 (TIMP-1) mRNA were studied by reverse transcription-polymerase chain reaction (RT-PCR) analysis in conjunction with computer-assisted image analysis. MMP-1 and TIMP-1 proteins were quantified by enzyme-linked immunosorbent assay (ELISA).

RESULTSEGCG decreased transcription activity of Jun protein after induction by UVA. Both the mRNA and protein levels of MMP-1 were increased by UVA irradiation, while no significant changes were observed in TIMP-1 levels. The ratio of MMP-1 to TIMP-1 showed statistically significant differences compared with the control. EGCG decreased the ratio of MMP-1 to TIMP-1 by inhibiting UVA-induced MMP-1 expression (P < 0.05).

CONCLUSIONEGCG can protect human fibroblasts against UVA damage by downregulating the transcription activity of Jun protein and the expression of MMP-1. The ratio of MMP-1 to TIMP-1, rather than the levels of MMP-1 or TIMP-1 alone, may play a significant role in human skin photodamage.

Catechin ; analogs & derivatives ; pharmacology ; Cells, Cultured ; Fibroblasts ; metabolism ; radiation effects ; Gene Expression Regulation ; drug effects ; Humans ; Matrix Metalloproteinase 1 ; biosynthesis ; genetics ; Proto-Oncogene Proteins c-jun ; analysis ; RNA, Messenger ; analysis ; Radiation-Protective Agents ; pharmacology ; Reverse Transcriptase Polymerase Chain Reaction ; Tissue Inhibitor of Metalloproteinase-1 ; biosynthesis ; genetics ; Ultraviolet Rays

OBJECTIVEIn order to investigate the roles of metalloproteinase in inflammatory bone destruction in ankylosing spondylitis (AS), and analyze the mechanism of preventing inflammatory bone destruction of Bushen Qiangdu decoction (BSQDD) in AS cases. Comparisons were made on the expressions of matrix metalloproteinase 9 (MMP-9) and tissue inhibitor of metalloproteinase 1 (TIMP-1) by peripheral blood mononuclear cells (PBMC) between AS patients and healthy controls. The effect of BSQDD was investigated on the expression and of MMP-9 and TIMP-1 produced by PBMC in AS patients.

METHODSFrom March 2005 to March 2006, 30 active AS cases of Kidney-asthenia, Du-cold and blood-stasis syndrome were selected as patients group in the China-Japan Friendship Hospital. There are 27 male patients and 3 female patients. The age range is from 16 to 45, averaging (30.8 +/- 8.8) years. Disease duration is from 0.5 to 10 years. Cases received three-month BSQDD treatment were considered as the treatment group. Twenty healthy persons were included in the control group. Serum and PBMC were separated. The PBMC were stimulated by PHA and PMA, and the supernatant was collected. The mRNA expression of MMP-9 and TIMP-1 in PBMC was analyzed by RT-PCR. The content of MMP-9 and TIMP-1 in serum and culture supernatant of PBMC were detected by ELISA.

RESULTSCompared with health control group, the serum concentration of MMP-9 and TIMP-1 in patients group before treatment increased (P<0.01, P<0.05), but the level of MMP-9 and TIMP-1 in the serum of patients after treatment decreased compared with pre-treatment cases (P<0.05). Furthermore,compared with health control group, PBMC of patients group before treatment expressed higher levels of MMP-9 and TIMP-1 both on transcript level and at protein level (P<0.01, P<0.05), and the expression levels of MMP-9 and TIMP-1 in PBMC in patients after treatment both on transcript level and at protein level was down-regulated compared with pre-treatment (P<0.01, P<0.05).

CONCLUSIONPBMC of AS patients had a higher potential capacity for MMP-9 and TIMP-1. BSQDD possibly prevented inflammatory bone destruction of AS through inhibiting production of MMP-9 and TIMP-1 produced by PBMC.

Adolescent ; Adult ; Case-Control Studies ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Gene Expression Regulation ; drug effects ; Humans ; Leukocytes, Mononuclear ; drug effects ; metabolism ; Male ; Matrix Metalloproteinase 9 ; biosynthesis ; blood ; genetics ; Middle Aged ; RNA, Messenger ; genetics ; metabolism ; Retrospective Studies ; Spondylitis, Ankylosing ; blood ; drug therapy ; genetics ; metabolism ; Tissue Inhibitor of Metalloproteinase-1 ; biosynthesis ; blood ; genetics ; Young Adult

PURPOSE: The degradation of the extracellular matrix has been shown to play an important role in the treatment of hepatic cirrhosis. In this study, the effect of thalidomide on the degradation of extracellular matrix was evaluated in a rat model of hepatic cirrhosis. MATERIALS AND METHODS: Cirrhosis was induced in Wistar rats by intraperitoneal injection of carbon tetrachloride (CCl4) three times weekly for 8 weeks. Then CCl4 was discontinued and thalidomide (100 mg/kg) or its vehicle was administered daily by gavage for 6 weeks. Serum hyaluronic acid, laminin, procollagen type III, and collagen type IV were examined by using a radioimmunoassay. Matrix metalloproteinase-13 (MMP-13), tissue inhibitor of metalloproteinase-1 (TIMP-1), and alpha-smooth muscle actin (alpha-SMA) protein in the liver, transforming growth factor beta1 (TGF-beta1) protein in cytoplasm by using immunohistochemistry and Western blot analysis, and MMP-13, TIMP-1, and TGF-beta1 mRNA levels in the liver were studied using reverse transcriptase polymerase chain reaction. RESULTS: Liver histopathology was significantly better in rats given thalidomide than in the untreated model group. The levels of TIMP-1 and TGF-beta1 mRNA and protein expressions were decreased significantly and MMP-13 mRNA and protein in the liver were significantly elevated in the thalidomide-treated group. CONCLUSION: Thalidomide may exert its effects on the regulation of MMP-13 and TIMP-1 via inhibition of the TGF-beta1 signaling pathway, which enhances the degradation of extracellular matrix and accelerates the regression of hepatic cirrhosis in rats.
Actins ; Animals ; Carbon Tetrachloride/toxicity ; Collagen Type III/metabolism ; Down-Regulation ; Extracellular Matrix/metabolism ; Immunohistochemistry ; Immunosuppressive Agents/*pharmacology ; Liver Cirrhosis, Experimental/chemically induced/*metabolism/pathology/*prevention & control ; Male ; RNA, Messenger/analysis/metabolism ; Rats ; Rats, Wistar ; Thalidomide/*pharmacology ; Tissue Inhibitor of Metalloproteinase-1/biosynthesis/*drug effects ; Transcription Factor RelA/biosynthesis/drug effects ; Transforming Growth Factor beta1/biosynthesis/*drug effects ; Transforming Growth Factors/metabolism