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Effect of thrombin on blood brain barrier permeability and its mechanism

Chinese Medical Journal 2004年第11期117卷 1677-1681页

作者：关景霞孙圣刚曹学兵陈志斌童萼塘 Department of Neurology Union Medical College Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430022 China Department of Neurology Affiliated Hospital of Hainan Medical College Haikou 570001 China

Background Previous studies have indicated that thrombi n (TM) may play a major role in brain edema after intracerebral hemorrhages (ICHs). However, the mechanism of TM-induced brain edema is poorly understood. In this study, we explored the effect of TM on the permeability of the blood brain barrier (BBB) and investigated its possible mechanism, aiming at providing a potential target for brain edema therapy after *** TM or TM + cathepsin G (CATG) was stereotaxically injected into the right caudate nucleus of Sprague-Dawley rats in vivo. BBB permeability was measured by Evans-Blue extravasation. Brain water content was determined by the dry-wet weight method. Brain microvascular endothelial cells were then cultured in vitro. After TM or TM+CATG was added to the endothelial cell medium, changes in the morphology of cells were dynamically observed by phase-contrast light microscopy, and the expression of matrix metalloproteinase-2 (MMP-2) protein was measured by immunohistochemical *** BBB permeability increased at 6 hours after a TM injection into the ipsilateral caudate nucleus (P24 hours (P0.05). TM induced endothelial cell contraction in vitro in a time-dependent manner and enhanced the expression of MMP-2 protein. After incubation with TM+CATG, cell morphology and MMP-2 expression did not change significantly as compared to the control group (P>0.05).Conclusions Increased BBB permeability may be one of the mechanisms behind TM-induced cerebral edema. TM induces endothelial cell contraction and promotes MMP-2 expression by activating protease activated receptor-1 (PAR-1), possibly leading to t

关键词： thrombin cerebral edema blood brai n barrier protease activated receptor-1 matrix metalloproteinase-2 cathepsin G

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Alendronate inhibits cell invasion and MMP-2 secretion in human chondrosarcoma cell line

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Acta Pharmacologica Sinica 2007年第8期28卷 1231-1235页

作者： Te-jen Lai Sheng-feng HSU Te-mao li Horng-chaung HSU Juang-geng lIn Chin-jung HSU Ming-chih CHOU Meng-chih LEE Shun-fa yang Yi-chin FONG Institute of Medicine Chung Shah Medical University~3Department of PsychiatryChung Shan Medical University HospitalTaiwanChina Gruduate Institute of Acupuncture Science China Medical UniversityTaiwanChina Department of Orthopaedic Surgery China Medical University HospitalTaiwanChina Institute of Medicine Chung Shah Medical University School of Chinese Medicine China Medical UniversityTaiwanChina

Aim:Chondrosarcoma is a malignant primary bone tumor that responds poorly toboth chemotherapy and radiation *** aim of the present study was toinvestigate the effect of alendronate,a bisphosphonate,on the invasion andmigration of human chondrosarcoma cells(JJ012).Methods:JJ012 cells weretreated with alendronate of various concentrations up to 100μmol/L for a speci-fied period,and then gelatin zymography and matrigel invasion assay was per-formed to study the effects of alendronate on matrix metalloproteinase(MMP)-2activity and the invasion ability of JJ012 cells,***:Our datashowed that alendronate exerted a dose-and time-dependent inhibitory effect onthe invasion and migration of JJ012 ***,gelatin zymography andRT-PCR showed that alendronate treatment decreased the activity and mRNAlevels of MMP-2 in a concentration-dependent ***:Our find-ings suggest that alendronate may reduce MMP-2 secretion at the transcriptionaland translational levels,and inhibit the invasion of chondrosarcoma ***,alendronate may be a potential candidate for the systemic therapy of chondro-sarcomas,as well as other malignant diseases.

关键词： alendronate" bisphosphonate chondrosarcoma matrix metalloproteinase-2 invasion migration

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Anti-proliferative effect of olmesartan on Tenon's capsule fibroblasts

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International Journal of Ophthalmology(English edition) 2016年第5期9卷 669-676页

作者： Xuan Wang Ya-Zhi Fan Liang Yao Jian-Ming Wang Department of Ophthalmology the Second Affiliated Hospital of Xi'an Jiaotong University

AIM： To evaluate the inhibitive effect of olmesartan to fibroblast proliferation and the anti-scarring effect in Tenon＇s capsule, both in vitro and in vivo.· METHODS： Human primary Tenon＇s capsule fibroblasts were cultured in vitro, treated with up titrating concentrations of olmesartan. The rate of inhibition was tested with methyl thiazol tetrazolium（MTT） ***-time PCR was performed to analyze changes in m RNA expressions of the fibrosis-related factors： matrix metalloproteinase-2（MMP-2）, tissue inhibitor of metalloproteinase（TIMP-1,2） and proliferating cell nuclear antigen（PCNA）. Thirty rabbits were divided into5 groups（3, 7, 14, 21, and 28d）. A rabbit conjunctiva flap model was created in each eye. Olmesartan solution was injected subconjunctivally and then evaluated its anti-proliferation and anti-fibrosis effects through the histological morphology and immunohistochemistry of MMP-2 and PCNA in each group. Only the 7d group was treated with Masson＇s trichrome to compare the neovascularization in the subconjunctiva area.·RESULTS： In vitro, cultured Tenon＇s capsule human fibroblasts showed a dose dependent inhibition by olmesartan in MTT. Olmesartan reduced m RNA expressions of MMP-2 and PCNA but increased m RNA expressions of TIMP-1 and TIMP-2. In vivo, the rabbit eyes treated with olmesartan at 3rd, 7th, 14 thand 21stdays demonstrated a significant reduced expressions of MMP-2 and PCNA compared with control eye, no significant difference observed in 28 thday group. The cellular proliferation and neovascularization was suppressed by olmesartan in Masson＇s trichrome observation.·CONCLUSION： By inhibiting fibroblasts in vitro and in vivo, olmesartan prevents the proliferation and activity of fibroblasts in scar tissue formation, which might benefit glaucoma filtering surgery.

关键词： olmesartan trabeculectomy anti-proliferative matrix metalloproteinase-2 proliferating cell nuclear antigen

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Retinoic Aacid Diminished the Expression of MMP-2 in Hyperoxia-exposed Premature Rat Lung Fibroblasts through Regulating Mitogen-activated Protein Kinases

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2011年第2期31卷 251-257页

作者：李文斌常立文容志惠刘伟 Department of Pediatrics Tongji Hospital Tongji Medical University Huazhong University of Science and Technology

This study examined the effects of retinoic acid （RA）, PD98059, SP600125 and SB203580 on the hyperoxia-induced expression and regulation of matrix metalloproteinase-2 （MMP-2） and metalloproteinase-2 （TIMP-2） in premature rat lung fibroblasts （LFs）. LFs were exposed to hyperoxia or room air for 12 h in the presence of RA and the kinase inhibitors PD98059 （ERK1/2）, SP600125 （JNK1/2） and SB203580 （p38） respectively. The expression levels of MMP-2 and TIMP-2 mRNA were detected by semi-quantitative reverse transcription polymerase chain reaction （RT-PCR）. MMP-2 activity was measured by zymography. The amount of p-ERK1/2, REK1/2, p-JNK1/2, JNK1/2, p-p38 and p38 was determined by Western blotting. The results showed that：（1） PD98059, SP600125 and SB203580 significantly inhibited p-ERK1/2, p-JNK1/2 and p-p38 respectively in LFs; （2） The expression of MMP-2 mRNA in LFs exposed to hyperoxia was decreased after treatment with RA, SP600125 and SB203580 respectively （P0.01 or 0.05）, but did not change after treatment with PD98059 （P0.05）. Meanwhile, RA, PD98059, SP600125 and SB203580 had no effect on the expression of TIMP-2 mRNA in LFs exposed to room air or hyperoxia （P0.05）; （3） The expression of pro- and active MMP-2 experienced no change after treatment with RA or SP600125 in LFs exposed to room air （P0.05）, but decreased remarkably after hyperoxia （P0.01 or 0.05）. SB203580 inhibited the expression of pro- and active MMP-2 either in room air or under hyperoxia （P0.01）. PD98059 exerted no effect on the expression of pro- and active MMP-2 （P0.05）. It was suggested that RA had a protective effect on hyperoxia-induced lung injury by down-regulating the expression of MMP-2 through decreasing the JNK and p38 activation in hyperoxia.

关键词： hyperoxia retinoic acid lung fibroblasts premature rats matrix metalloproteinase-2 mitogen-activated protein kinases

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Involvement of MMP-2 in Adriamycin Resistance Dependent on ERK1/2 Signal Pathway in Human Osteosarcoma MG-63 Cells

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2012年第1期32卷 82-86页

作者：任晔郭风劲陈安民邓睿王江 Department of Orthopedics Tongji HospitalTongji Medical CollegeHuazhong University of Science and Technology

matrix metalloproteinase-2 (MMP-2) level and the ERK1/2 signal pathway are dependent factors for the growth and metastasis of ***,the impact of MMP-2 in combination with ERK1/2 in tumor patients with drug resistance is *** determine the relationship between MMP-2 and the ERK1/2 signal pathway,we established an adriamycin (ADM)-induced MG-63 (ADM-MG-63) cell *** the increase of the ERK1/2 pathway blocker PD98059,we detected the expression levels of MMP-2 and p-ERK1/2 by Western blot in ADM-MG-63 *** ADM-MG-63 cells transfected with MMP-2-siRNA,the expression of ERK1/2 was detected for understanding the function of the ERK1/2 signal *** siRNAs for MMP-2 (MMP-2-siRNA) were designed,and the optimal one was selected and tested at different time points of 24,48 and 72 *** an ADM-induced condition,ADM-MG-63 cells were finally stable living in the medium of ADM (200 ng/mL).PD98059 could effectively suppress the expression levels of p-ERK1/2 and *** the MMP-2 was silenced by using MMP-2-siRNA,the expression of p-ERK1/2 was *** is con-cluded that MMP-2 may be involved in ADM resistance dependent on ERK1/2 signal pathway,sug-gesting interference in ERK1/2 may be a new method of targeted therapy for tumor resistance.

关键词： adriamycin matrix metalloproteinase-2 osteosarcoma RNA interference

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Expression of MMP-2 and MMP-9 in retinoblastoma and their significance

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International Journal of Ophthalmology(English edition) 2011年第5期4卷 489-491页

作者： Hua Long, Fa-Gang Jiang Department of Ophthalmology Suizhou Central Hospital Suizhou 441322 Hubei Province China Department of Ophthalmology Union Hospital of Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 Hubei Province China

AIM: To investigate the expression of matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) in retinoblastoma (Rb), and their relationships with tumor development stage. METHODS: Immunohistochemical technique was used to detect the expression of MMP-2 and MMP-9 in 41 cases of paraffin embedded Rb samples. Quantitative analysis of the expression of MMP-2 and MMP-9 was assessed by HMIAS-2000 Color Pathologic Analysis System. The differences of the expression of MMP-2 and MMP-9 in each clinical and pathological stage were analyzed statistically. RESULTS: In all the 41 Rb specimens, MMP-2 and MMP-9 expression was found in tumor cells. The expression of MMP-2 and MMP-9 was significantly higher in tumors with optic nerve invasion than in tumors without optic nerve invasion (P<0.05); the expression of MMP-2 and MMP-9 was significantly higher in tumors of extra-ocular stage than in tumors of glaucomatous stage or intra-ocular stage( P<0.05). CONCLUSION: MMP-2 and MMP-9 exist in retinoblastoma cells. The level of MMP-2 and MMP-9 is related to optic nerve invasion and clinical stage of Rb, which suggests the expression of MMP-2 and MMP-9 could be connected to the invasion and development of tumor cells. Further research is needed for deeper understanding of the biological behavior and better evaluation of the prognosis of Rb.

关键词： etinoblastoma matrix metalloproteinase-2 matrix metalloproteinase-9 immunohistochemistry

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M-CSF TARGETING INTO LCL NUCLEUS BEHAVES AS A MALIGNANCY PROMOTOR

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Chinese Journal of Cancer Research 2003年第4期15卷 262-268页

作者：曹震宇吴克复宋玉华李戈林永敏饶青马小彤 State Key Laboratory for Experimental Hematology Institute of Hematology Chinese Academy of Medical Sciences & Peking Union Medical College Tianjin 300020

Objective: To investigate the functions of nM-CSF in malignant cells. Methods: recombinant M-CSF was targeted into cell nucleus by employing a eukaryotic expression plasmid vector pCMV/myc/nuc. The constructed plasmid was transfected into cells of EBV transformed lymphoblastoid cell line (LCL). RT-PCR, Western blot and immunofluorescent staining showed that recombinant M-CSF was localized into LCL cell nucleus. The transgenic cells showed elevated proliferation potential, enhanced resistance to apoptosis and increased ability of in vitro migration. Conclusion: Nucleus presenting M-CSF might act as a promoting factor in the processes of cell malignancy.

关键词： Macrophage colony-stimulating factor Nuclear localization sequence Lymphoblastoid cell line (LCL) matrix metalloproteinase-2 Apoptosis

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Expression of MMP-2 in residual VX2 liver tumor after transcatheter arterial embolization combined with portal venous embolization in an animal model

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Journal of Interventional Medicine 2020年第4期3卷 167-173页

作者： Ning Wei Ziqiang Wu Dong Lu Jingkun Xiao Chunze Zhou Senlin Chu Weifu Lv Department of Radiology The First Affiliated Hospital of USTCDivision of Life Sciences and MedicineUniversity of Science and Technology of ChinaHefeiAnhui230001China Department of Medical Imaging Center The First Affiliated Hospital of USTCDivision of Life Sciences and MedicineUniversity of Science and Technology of ChinaHefeiAnhui230036China

Objective:This study aimed to analyze the effects of transcatheter arterial embolization(TAE)combined with portal venous embolization(PVE)on the expression of MMP-2 in residual VX2 liver tumor tissues,liver function and non-embolic lobe ***:A total of 72 rabbits were randomly divided into Sham,TAE,PVE and TAE+PVE groups(n=18/group).The tissue samples from each group were taken at 6 h,3 days and 7 days after interventional operation,***-2 expression was detected by immunohistochemistry,Real-time PCR,and *** main indicators(such as AST,ATL,and TBIL)of liver function and the volume of non-embolized hepatic lobes were measured in each group after ***-way ANOVA and Kruskal-wallis method were used for statistical ***:The expression of MMP-2 mRNA and protein remained the highest in the Sham group,and the expression of MMP-2 mRNA and protein in TAE,PVE and TAE+PVE groups were successively increased,and the expression of MMP-2 in TAE+PVE group was always significantly higher than TAE *** AST and ALT levels in each group on day 7 after operation showed a significant declination,and all groups have recovered to the preoperative baseline level and TBIL has a slight fluctuation in each group after operation with no statistical *** day 7 after operation,the increasing volume of non-embolized liver lobes in TAE+PVE group showed a more significant effect than those in PVE group,but there was no statistical significance(37.62±1.54 ml VS 36.18±1.15 ml,P=0.881),and its volume was significantly higher than those in the sham group(27.03±1.11 ml).Conclusion:TAE+PVE is considered to be an efficient and safe approach for treating rabbit VX2 liver transplantation tumor,but the expression of MMP-2 increased fastest after TAE+PVE,which might promote tumor cell invasion and metastasis.

关键词： Transcatheter arterial embolization Portal vein embolization matrix metalloproteinase-2 VX2 tumor Liver regeneration

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The ratio of MMP-2 to TIMP-2 in hilar cholangiocarcinoma:a semi-quantitative study

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Hepatobiliary & Pancreatic Diseases International 2004年第4期3卷 599-602页

作者： Mei Xiao, Nin-Xing Zhou,Zhi-Qiang Huang,Ya-Li Lu, Ling-Hong Chen, Dian-Jun Wang and Wei-Lin Chang Beijing, China Department of Hepatobiliary Surgery, General Air- force of Hospital, Beijing 100036, China and General Hospital of People’ s Liberation Army, Beijing 100853 , China Department of Hepatobiliary Surgery of General Airforce of Hospital Beijing 100036 China. fengweiairforce@***

BACKGROUND: Hilar cholangiocarcinoma is associated with low resectability and poor survival. The aim of this study was to evaluate the roles of matrix metalloproteinases- 2 (MMP-2 ) and its tissue inhibitor of metalloproteinase-2 (TIMP-2) in tumor invasion or as a prognostic factor in patients with human hilar cholangiocarcinoma. METHODS: The expressions of MMP-2 and TIMP-2 were investigated in patients. Paraffinized tissue sections ob- tained from 50 patients with human hilar cholangiocarcino- ma were analysed. The expressions of MMP-2 and TIMP-2 were examined immunohistochemically. Image analysis with image-pro plus analysis software was used to semi- quantitatively determine the ratio of MMP-2 to TIMP-2. RESULTS: The expression levels of MMP-2 and TIMP-2 were strongly associated with tumor hepatic invasion in pa- tients with hilar cholangiocarcinoma. Significant diffe- rences in the ratio of MMP-2 to TIMP-2 between some pathologic factors were observed in patients with hilar cholangiocarcinoma. CONCLUSIONS: MMP-2 plays an essential role in tumor invasion and metastasis,while TIMP-2 is shown to strongly inhibit cancer invasion and metastasis. The ratio of MMP-2 to TIMP-2 may be a prognostic indicator for patients with hilar cholangiocarcinoma.

关键词： hilar cholangiocarcinoma matrix metalloproteinase-2 tissue inhibitor of metalloproteinase-2 immunohistochemical staining tumor invasion prognostic factor

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Fuzheng Huayu Formula(扶正化瘀方) Prevents Rat Renal Interstitial Fibrosis Induced by HgCl2 via Antioxidative Stress and Down-regulation of Nuclear Factor-Kappa B Activity

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Chinese Journal of Integrative Medicine 2017年第8期23卷 598-604页

作者： YUAN Ji-li TAO Yan-yan WANG Qing-lan SHEN Li LIU Cheng-hai Institute of Liver Diseases Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine Shangha 201203 China Shanghai Key Laboratory of Traditiona Chinese Clinical Medicine Shanghai 201203 China E-Institute of TCM Internal Medicine Shanghai Municipa Education Commission Shanghai 201203 China

Objective：To investigate the mechanism of action of Fuzheng Huayu Formula（扶正化瘀方,FZHY）against renal interstitial fibrosis（RIF）relating to oxidative injury and nuclear factor-kappa B（NF-κB）***：Thirty-two Sprague-Dawley rats were randomly divided into 3 groups：normal group,model group and FZHY treatment *** RIF model was induced by oral administration of HgC l2 at a dose of 8 mg/kg body weight once a day for 9 ***,rats in FZHY treatment group orally took FZHY at a dose of4.0 g/kg rat weight for 9 *** content of hydroxyproline（Hyp）and collagen deposition in kidney were *** activities of superoxide dismutase（SOD）and glutathione peroxidase（GSH-Px）,the content of glutathione（GSH）and malondialdehyde（MDA）of kidney were *** expressions of inhibitor-κappa B（IκB）,phospho-IκB（p-IκB）,tumor necrosis factor-α（TNF-α）,matrix metalloproteinase-2（MMP-2）andα-smooth muscle actin（α-SMA）were analyzed by Western blot.α-SMA expression was also observed by immunofluorescent ***-2 activity was measured by gelatin ***-κB activation was determined by electrophoretic mobility shift ***：Renal interstitial fibrosis was induced by Hg Cl2,demonstrated by remarkably increased Hyp contents and excessive collagen deposition in kidney（P〈0.01）.FZHY significantly inhibited renal interstitial collagen deposition and reduced Hyp content of the Hg Cl2-treated rats（P〈0.01）.GSH content decreased obviously,and MDA content increased significantly in HgC l2-treated rats compared with that of normal rats（P〈0.01）.FZHY significantly increased GSH content and decreased MDA content in the model rats（P〈0.01）.The expressionα-SMA was increased in model rats compared with that of normal rats,FZHY significantly decreased its expression（P〈0.01）.The expressions of p-IκB and TNF-αand MMP-2,MMP-2 activity,and NF-κB activation were increased in model group compared with that in normal group（P〈0.01）,FZHY significantly decreased NF-κB activation,MMP-

关键词： Fuzheng Huayu Formula renal interstitial fibrosis mercuric chloride oxidative stress nuclearfactor-kappa B matrix metalloproteinase-2 tumor necrosis factor- α Chinese medicine

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