OBJECTIVE: To explore the expression and distribution of a disintegrin and metalloprotease with thrombospondin motif (ADAMTS)-2 and transforming growth factor (TGF) -β1 in patients with or without cirrhosis, and to determine their relation. METHODS: The liver tissues from 16 patients with cirrhotic portal hypertensive and 8 patients with liver injury were collected in Wuhan General Hospital from March to June, 2010. Immunohistochemistry and Western blot were applied to detect the protein expression of ADAMTS-2 and TGF-β1. RESULTS: Immunohistochemistry showed that the expression of ADAMTS-2 and TGF-β1 was significantly higher in the cirrhotic tissues than that in normal tissues (P<0.05). Western blot also showed the expression of ADAMTS-2 and TGF-β1 in the cirrhosis tissues was significantly higher than that in normal tissues (P<0.05). There was a positive correlation between ADAMTS-2 and TGF-β1 (r=0.862, P<0.01). CONCLUSION ADAMTS-2 and TGF-β1 may have a synergistic reaction in promoting liver cirrhosis.
ADAM Proteins ; metabolism ; ADAMTS Proteins ; ADAMTS4 Protein ; Blotting, Western ; Humans ; Immunohistochemistry ; Liver ; metabolism ; pathology ; Liver Cirrhosis ; metabolism ; Procollagen N-Endopeptidase ; metabolism ; Transforming Growth Factor beta1 ; metabolism

OBJECTIVETo study the effect of RNA interference (RNAi)-mediated aggrecanase-1 gene silencing on extracellular matrix metabolism of cultured rat costochondral chondrocytes.

METHODSRat costochondral chondrocyte monolayers were obtained by microdissection and digestion. The growth and morphological changes of the chondrocytes were observed after RNAi of aggrecanase-1 gene. The mRNA expression of aggrecanase-1 was detected by RT-PCR method, and aggrecan content was determined by Western blotting.

RESULTSThe specific inhibition of aggrecanase-1 by RNAi produced no adverse effect on the morphology and growth of the chondrocytes. The mRNA of aggrecanase-1 decreased and aggrecan content increased significantly after transfection of the chondrocytes.

CONCLUSIONInhibition of aggrecanase-1 decreases aggrecan degradation in cultured rat chondrocytes. RNAi technique can be a useful means for studying extracellular matrix metabolism in the cartilage.

ADAM Proteins ; genetics ; metabolism ; ADAMTS4 Protein ; Aggrecans ; metabolism ; Animals ; Cells, Cultured ; Chondrocytes ; cytology ; metabolism ; Extracellular Matrix ; metabolism ; Female ; Procollagen N-Endopeptidase ; genetics ; metabolism ; RNA Interference ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Transfection

BACKGROUND/AIMS: SKI306X, a mixed extract of three herbs, Clematis mandshurica (CM), Prunella vulgaris (PV), and Trichosanthes kirilowii (TK), is chondroprotective in animal models of osteoarthritis (OA). The objectives of this study were to investigate its effect on interleukin (IL)-1beta-induced degradation of glycosaminoglycan (GAG) and the basis of its action in human OA cartilage, as well as to screen for the presence of inhibitors of matrix metalloproteinase (MMP)-13 and a disintegrin and metalloprotease with thrombospondin motifs (ADAMTS)-4 in SKI306X and its component herbs, as well as in fractions from SKI306X. METHODS: Human OA chondrocytes and cartilage explants were obtained during total knee replacements and incubated with IL-1beta +/- oncostatin M with or without SKI306X or its component herb extracts. GAG degradation was assayed in cartilage explants using a commercial kit. Expression of genes involved in cartilage destruction was measured by real-time polymerase chain reaction using chondrocyte RNA. SKI306X was fractionated by preparative liquid chromatography to test for the presence of inhibitors of MMP-13 and ADAMTS-4. RESULTS: SKI306X and PV inhibited IL-1beta-induced GAG release from cartilage explants, and SKI306X, CM, PV, and TK inhibited IL-1beta-induced MMP gene expression. Unexpectedly, SKI306X greatly stimulated IL-1beta + oncostatin M-induced ADAMTS-4 gene expression, probably due to its TK component. Some fractions of SKI306X also inhibited ADAMTS-4 activity. CONCLUSIONS: SKI306X and its herbal components inhibit GAG degradation and catabolic gene expression in human OA chondrocytes and cartilage explants. SKI306X likely also contains one or more ADAMTS-4 inhibitor.
ADAM Proteins/antagonists & inhibitors ; Cartilage, Articular/*drug effects/*metabolism ; Cells, Cultured ; Chondrocytes/drug effects/metabolism ; Down-Regulation/drug effects ; Drugs, Chinese Herbal/*pharmacology ; Glycosaminoglycans/*metabolism ; Humans ; Interleukin-1beta/metabolism ; Matrix Metalloproteinase 13/metabolism ; Matrix Metalloproteinase Inhibitors/pharmacology ; Oncostatin M/metabolism ; Osteoarthritis, Knee/drug therapy/genetics/metabolism ; Procollagen N-Endopeptidase/antagonists & inhibitors