Inhibiting MMP13 Attenuates Deep Vein Thrombosis in a Mouse Model by Reducing the Expression of Pdpn

Objective:Matrix metalloproteinase 13(MMP13)is an extracellular matrix protease that affects the progression of atherosclerotic plaques and arterial thrombi by degrading collagens,modifying protein structures and regulating inflammatory responses,but its role in deep vein thrombosis(DVT)has not been determined.The purpose of this study was to investigate the potential effects of MMP13 and MMP13-related genes on the formation of DVT.Methods:We altered the expression level of MMP13 in vivo and conducted a transcriptome study to examine the expression and relationship between MMP13 and MMP13-related genes in a mouse model of DVT.After screening genes possibly related to MMP13 in DVT mice,the expression levels of candidate genes in human umbilical vein endothelial cells(HUVECs)and the venous wall were evaluated.The effect of MMP13 on platelet aggregation in HUVECs was investigated in vitro.Results:Among the differentially expressed genes,interleukin 1 beta,podoplanin(Pdpn),and factor VIII von Willebrand factor(F8VWF)were selected for analysis in mice.When MMP13 was inhibited,the expression level of PDPN decreased significantly in vitro.In HUVECs,overexpression of MMP13 led to an increase in the expression level of PDPN and induced platelet aggregation,while transfection of PDPN-siRNA weakened the ability of MMP13 to increase platelet aggregation.Conclusions:Inhibiting the expression of MMP13 could reduce the burden of DVT in mice.The mechanism involves downregulating the expression of Pdpn through MMP13,which could provide a novel gene target for DVT diagnosis and treatment.

Salvia miltiorrhiza-asarum ointment combined with Chinese medical massage alleviates symptoms of osteoarthritis in a rat model through the Notch1/matrix metalloproteinase-13 signaling pathway

OBJECTIVE:This study investigated the effect of salvia miltiorrhiza-asarum ointment(SMAO)plus Chinese medical massage on knee osteoarthritis in a rat model.METHODS:Hulth's method was used to establish a Sprague-Dawley rat model of knee osteoarthritis(OA).The levels of matrix metalloproteinase-13(MMP-13),collagen-Ⅱ,aggrecan,interleukin(IL)-1β,tumor necrosis factor-α(TNF-α),and IL-6 were measured by enzyme-linked immunosorbent assays.The joint space was assessed by a Perlove X-ray system.Histopathology was examined by hematoxylin-eosin and Safranin O staining.The mRNA and protein expression levels of Notch1,MMP-13,collagen-Ⅱ,and aggrecan were measured by quantitative reverse transcription-polymerase chain reaction and Western blotting,respectively.RESULTS:SMAO plus Chinese medical massage significantly decreased the levels of MMP-13,IL-1β,TNF-α,and IL-6,and increased serum collagen-Ⅱ and aggrecan levels.Pathological injury of the knee joint was improved by SMAO treatment.mRNA and protein expression of Notch1 and MMP-13 was remarkably downregulated,but collagen-Ⅱ and aggrecan levels were significantly upregulated in cartilage tissues.CONCLUSION:SMAO combined with Chinese medical massage effectively relieves OA symptoms,which may involve inhibiting inflammation through the Notch1/MMP-13 signaling pathway.

Effects of HDAC4 on IL-1β-induced matrix metalloproteinase expression regulated partially through the WNT3A/β-catenin pathway

Background::Histone deacetylase 4(HDAC4)regulates chondrocyte hypertrophy and bone formation.The aim of the present study was to explore the effects of HDAC4 on Interleukin 1 beta(IL-1β)-induced chondrocyte extracellular matrix degradation and whether it is regulated through the WNT family member 3A(WNT3A)/β-catenin signaling pathway.Methods::Primary chondrocytes(CC)and human chondrosarcoma cells(SW1353 cells)were treated with IL-1βand the level of HDAC4 was assayed using Western blotting.Then,HDAC4 expression in the SW1353 cells was silenced using small interfering RNA to detect the effect of HDAC4 knockdown on the levels of matrix metalloproteinase 3(MMP3)and MMP13 induced by IL-1β.After transfection with HDAC4 plasmids,the overexpression efficiency was examined using Real-time quantitative polymerase chain reaction(qRT-PCR)and the levels of MMP3 and MMP13 were assayed using Western blotting.After incubation with IL-1β,the translocation ofβ-catenin into the nucleus was observed using immunofluorescence staining in SW1353 cells to investigate the activation of the WNT3A/β-catenin signaling pathway.Finally,treatment with WNT3A and transfection with glycogen synthase kinase 3 beta(GSK3β)plasmids were assessed for their effects on HDAC4 levels using Western blotting.Results::IL-1βdownregulated HDAC4 levels in chondrocytes and SW1353 cells.Furthermore,HDAC4 knockdown increased the levels of MMP3 and MMP13,which contributed to the degradation of the extracellular matrix.Overexpression of HDAC4 inhibited IL-1β-induced increases in MMP3 and MMP13.IL-1βupregulated the levels of WNT3A,and WNT3A reduced HDAC4 levels in SW1353 cells.GSK-3βrescued IL-1β-induced downregulation of HDAC4 in SW1353 cells.Conclusion::HDAC4 exerted an inhibitory effect on IL-1β-induced extracellular matrix degradation and was regulated partially by the WNT3A/β-catenin signaling pathway.

Design, Synthesis and Evaluation of 6-Oxo-1, 6-dihydropyrimidine- 2,5-dicarboxamide Derivatives as MMP 13 Inhibitors

We designed and synthsized a series of novel 6-oxo-l,6-dihydropyrimidine-2,5-dicarboxamide derivatives and evaluated their inhibition effects on MMP 3, MMP 12 and MMP 13. The pharmacological results show that they have potent and highly selective activity of inhibiting MMP 13.

Modified Xiaochaihu Decoction(加味小柴胡汤) Promotes Collagen Degradation and Inhibits Pancreatic Fibrosis in Chronic Pancreatitis Rats

Objective:To investigate the effect of Modified Xiaochaihu Decoction(MXD,加味小柴胡汤)on collagen degradation in rats with chronic pancreatitis(CP).Methods:Rats were injected dibutyltin dichloride(DBTC,7 mg/kg of body weight)into the right caudal vein to induce CP model.Thirty heallhy male Wistar rats were randomly divided into three groups by a random number table:the control,the model and the treatment groups.Rats of treatment group were administered MXD(10 g/kg of body weight)orally once daily starting from the day post-model establishment.Pancreatic tissues were harvested after 28-day feeding and fibrosis was evaluated by picro-sirius red staining.The contents of collagen typeⅠandⅢwere detected using enzymelinked immunosorbent assay(ELISA),the expression of matrix metalloproteinase 13(MMP13)and tissue inhibitor of metalloproteinase 1(TIMP1)was analyzed by Western blot and real-time polymerase chain reaction(PCR).Results:The fibrosis scoring of pancreatic tissues,the concentrations of collagen typeⅠandⅢ,the expression levels of MMP13 and TIMP1 proteins and mRNA in the model group were all increased compared with the control group(P<0.05).After treatment with MXD,the fibrosis scoring of pancreatic tissues,the concentrations of collagen typeⅠandⅢ,the expression levels of MMP13 proteins and m RNA in the teatment group were all decreased compared with the model group(P<0.05),but there were no significant differences in the expression levels of TIMP1 proteins and m RNA(P>0.05).Conclusion:MXD could promote collagen degradation and reverse pancreatic fibrosis in CP rats via a mechanism involve up-regulation of MMP13 expression.