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 regu...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.展开更多
BACKGROUND: Interstitial collagenase has been considered as an essential enzyme for collagenolysis in liver fi-brosis, because type Ⅰ and Ⅲ collagens increase predominantly in liver fibrosis. The present study aimed...BACKGROUND: Interstitial collagenase has been considered as an essential enzyme for collagenolysis in liver fi-brosis, because type Ⅰ and Ⅲ collagens increase predominantly in liver fibrosis. The present study aimed to demonstrate the gene expression of matrix metalloproteinases-13 (MMP-13) in the progressive phases of ethanol induced experimental liver fibrosis in rats. METHODS: Thirty-four Sprague-Dawley rats were randomly divided into two groups. The experimental group (24 rats) was given ethanol (44% , 7 g/kg) every day and the control group (10) was given normal saline. Liver samples were harvested from experimental rats at 4, 12 and 24 weeks respectively. The kinetics of MMP-13 mRNA expression was assayed by semi-quantity reverse transcriptase-poly-merase chain reaction (RT-PCR). RESULTS: In normal rat liver, a faint band for MMP-13 mRNA was observed by RT-PCR (0.24±0.41). The gene expression of MMP-13 was increased in the liver of the rats treated with ethanol for 4 weeks (0. 62 ±0. 54), but it was not considered statistically significant (P >0.05). And the livers from 12-week-treated rats showed a marked mRNA expression(1.65 ±0.47, P <0. 01). Once fibrosis became prominent (24 weeks), a faint band of MMP-13 mRNA was observed (0.39±0.25). CONCLUSION: MMP-13 participates in the degradation of newly-formed matrix in the early phase of rat liver fibrosis induced by ethanol, and it was induced in a distinct time frame.展开更多
AIM:To examine the association between the single nucleotide polymorphisms (SNPs)of matrix metalloprotease-9 (MMP-9) gene and primary angleclosure glaucoma(PACG)in a Chinese Han population.METHODS:DNA samples were ext...AIM:To examine the association between the single nucleotide polymorphisms (SNPs)of matrix metalloprotease-9 (MMP-9) gene and primary angleclosure glaucoma(PACG)in a Chinese Han population.METHODS:DNA samples were extracted from peripheral-blood mononuclear cells of 214 PACG patients and 224 healthy controls.Genotyping of rs3918249,rs3918254,rs17577 and rs3787268 in MMP-9was performed using polymerase chain reaction restriction fragment length polymorphism(PCR-RFLP)analysis and the direct sequencing technique.The association between these genetic polymorphisms and risk of PACG was estimated by χ2 test.RESULTS:The distributions of rs3918249,rs3918254,rs17577 and rs3787268 genotypes among cases and healthy controls were compatible with that from HardyWeinberg equilibrium(HWE,P】0.05).The increased frequency of CC and CT genotypes of rs3918254 were observed in PACG patients compared to healthy controls[P=0.006,P corrected(Pcorr)=0.048].The haplotype analysis showed that the CCGG haplotype was nominal associated with PACG(P=0.015),however,the significant was lost when the Bonferroni correction was used(Pcorr=0.105).CONCLUSION:Our results revealed that rs3918254 in MMP-9 may be a susceptible locus to PACG in China,people with the CC and CT genotypes of rs3918254 are more susceptible to PACG.The susceptibility to PACG inChinese Han patients may be not influenced by SNPs rs3918249,rs3787268 and rs17577 in MMP-9.展开更多
Identification and quantification of low abundance growth factors and regulators in complex biological samples still present a challenging task in analytical biochemistry. Immunoassays are often used for such purpose ...Identification and quantification of low abundance growth factors and regulators in complex biological samples still present a challenging task in analytical biochemistry. Immunoassays are often used for such purpose but immunoassays face limitation of both availability and qualities of antibody reagents that are necessary for development of immune assays. With genomics data base available, mass spectrometry (MS) can analyze protein tryptic peptides directly for quantitative determination of proteins. In this study, we report a method for detection of matrix metalloproteinase 1 (MMP1), an important extracellular matrix modulator, in human breast cancer cells by quadrupole time-of-flight (Q-TOF) MS. Absolute quantification of MMP1 was conducted using the selected reaction monitoring (SRM) on a triple quadrupole (Triple-Quad) MS via transitions selected from MMP1 tryptic peptides using non isotope labeled MMP1 protein as a titration standard. In comparison with immune based assay, this MS method showed picogram level sensitivity for quantitative determination of MMP1 intotal cell lysates. Our results demonstrated the feasibility of absolute quantification of low abundance proteins using label-free protein standard by mass spectrometry. Therefore, this method provides not only advantages of high sensitivity but also cost saving in comparison with the commonly used mass spectrometry that currently employs isotype labeled proteins for quantitative analysis.展开更多
At present,there are no resto rative therapies in the clinic for spinal cord injury,with current treatments offering only palliative treatment options.The role of matrix metalloproteases is well established in spinal ...At present,there are no resto rative therapies in the clinic for spinal cord injury,with current treatments offering only palliative treatment options.The role of matrix metalloproteases is well established in spinal cord injury,howeve r,translation into the clinical space was plagued by early designs of matrix metalloprotease inhibitors that lacked specificity and fears of musculos keletal syndrome prevented their further development.Newe r,much more specific matrix metalloprotease inhibitors have revived the possibility of using these inhibitors in the clinic since they are much more specific to their to rget matrix metalloproteases.Here,the evidence for use of matrix metalloproteases after spinal cord injury is reviewed and researche rs are urged to overcome their old fears rega rding matrix metalloprotease inhibition and possible side effects for the field to progress.Recently published work by us shows that inhibition of specific matrix metalloproteases after spinal cord injury holds promise since four key consequences of spinal cord injury could be alleviated by specific,next-gene ration matrix metalloprotease inhibitors.For example,specific inhibition of matrix metalloprotease-9 and matrix metalloprotease-12 within 24 hours after injury and for 3 days,alleviates spinal cord injury-induced edema,blood-s pinal co rd barrier breakdown,neuro pathic pain and resto res sensory and locomotor function.Attempts are now underway to translate this therapy into the clinic.展开更多
基金supported by grants from General Project of Yunnan Basic Research Program(No.202301AT070104)the Joint Project of Kunming Medical University and Science and Technology Department of Yunnan Province(No.202001AY070001-185)+1 种基金the Joint Project of Kunming Medical University and Science and Technology Department of Yunnan Province(No.202101AY070001-119)Yunnan Provincial Orthopedic and Sports Rehabilitation Clinical Medicine Research Center(No.202102AA310068).
文摘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.
文摘BACKGROUND: Interstitial collagenase has been considered as an essential enzyme for collagenolysis in liver fi-brosis, because type Ⅰ and Ⅲ collagens increase predominantly in liver fibrosis. The present study aimed to demonstrate the gene expression of matrix metalloproteinases-13 (MMP-13) in the progressive phases of ethanol induced experimental liver fibrosis in rats. METHODS: Thirty-four Sprague-Dawley rats were randomly divided into two groups. The experimental group (24 rats) was given ethanol (44% , 7 g/kg) every day and the control group (10) was given normal saline. Liver samples were harvested from experimental rats at 4, 12 and 24 weeks respectively. The kinetics of MMP-13 mRNA expression was assayed by semi-quantity reverse transcriptase-poly-merase chain reaction (RT-PCR). RESULTS: In normal rat liver, a faint band for MMP-13 mRNA was observed by RT-PCR (0.24±0.41). The gene expression of MMP-13 was increased in the liver of the rats treated with ethanol for 4 weeks (0. 62 ±0. 54), but it was not considered statistically significant (P >0.05). And the livers from 12-week-treated rats showed a marked mRNA expression(1.65 ±0.47, P <0. 01). Once fibrosis became prominent (24 weeks), a faint band of MMP-13 mRNA was observed (0.39±0.25). CONCLUSION: MMP-13 participates in the degradation of newly-formed matrix in the early phase of rat liver fibrosis induced by ethanol, and it was induced in a distinct time frame.
基金Supported by National Natural Science Foundation of China(No.81270990No.81070723)
文摘AIM:To examine the association between the single nucleotide polymorphisms (SNPs)of matrix metalloprotease-9 (MMP-9) gene and primary angleclosure glaucoma(PACG)in a Chinese Han population.METHODS:DNA samples were extracted from peripheral-blood mononuclear cells of 214 PACG patients and 224 healthy controls.Genotyping of rs3918249,rs3918254,rs17577 and rs3787268 in MMP-9was performed using polymerase chain reaction restriction fragment length polymorphism(PCR-RFLP)analysis and the direct sequencing technique.The association between these genetic polymorphisms and risk of PACG was estimated by χ2 test.RESULTS:The distributions of rs3918249,rs3918254,rs17577 and rs3787268 genotypes among cases and healthy controls were compatible with that from HardyWeinberg equilibrium(HWE,P】0.05).The increased frequency of CC and CT genotypes of rs3918254 were observed in PACG patients compared to healthy controls[P=0.006,P corrected(Pcorr)=0.048].The haplotype analysis showed that the CCGG haplotype was nominal associated with PACG(P=0.015),however,the significant was lost when the Bonferroni correction was used(Pcorr=0.105).CONCLUSION:Our results revealed that rs3918254 in MMP-9 may be a susceptible locus to PACG in China,people with the CC and CT genotypes of rs3918254 are more susceptible to PACG.The susceptibility to PACG inChinese Han patients may be not influenced by SNPs rs3918249,rs3787268 and rs17577 in MMP-9.
文摘Identification and quantification of low abundance growth factors and regulators in complex biological samples still present a challenging task in analytical biochemistry. Immunoassays are often used for such purpose but immunoassays face limitation of both availability and qualities of antibody reagents that are necessary for development of immune assays. With genomics data base available, mass spectrometry (MS) can analyze protein tryptic peptides directly for quantitative determination of proteins. In this study, we report a method for detection of matrix metalloproteinase 1 (MMP1), an important extracellular matrix modulator, in human breast cancer cells by quadrupole time-of-flight (Q-TOF) MS. Absolute quantification of MMP1 was conducted using the selected reaction monitoring (SRM) on a triple quadrupole (Triple-Quad) MS via transitions selected from MMP1 tryptic peptides using non isotope labeled MMP1 protein as a titration standard. In comparison with immune based assay, this MS method showed picogram level sensitivity for quantitative determination of MMP1 intotal cell lysates. Our results demonstrated the feasibility of absolute quantification of low abundance proteins using label-free protein standard by mass spectrometry. Therefore, this method provides not only advantages of high sensitivity but also cost saving in comparison with the commonly used mass spectrometry that currently employs isotype labeled proteins for quantitative analysis.
文摘At present,there are no resto rative therapies in the clinic for spinal cord injury,with current treatments offering only palliative treatment options.The role of matrix metalloproteases is well established in spinal cord injury,howeve r,translation into the clinical space was plagued by early designs of matrix metalloprotease inhibitors that lacked specificity and fears of musculos keletal syndrome prevented their further development.Newe r,much more specific matrix metalloprotease inhibitors have revived the possibility of using these inhibitors in the clinic since they are much more specific to their to rget matrix metalloproteases.Here,the evidence for use of matrix metalloproteases after spinal cord injury is reviewed and researche rs are urged to overcome their old fears rega rding matrix metalloprotease inhibition and possible side effects for the field to progress.Recently published work by us shows that inhibition of specific matrix metalloproteases after spinal cord injury holds promise since four key consequences of spinal cord injury could be alleviated by specific,next-gene ration matrix metalloprotease inhibitors.For example,specific inhibition of matrix metalloprotease-9 and matrix metalloprotease-12 within 24 hours after injury and for 3 days,alleviates spinal cord injury-induced edema,blood-s pinal co rd barrier breakdown,neuro pathic pain and resto res sensory and locomotor function.Attempts are now underway to translate this therapy into the clinic.
