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BACKGROUND:At present, there is no effective treatment strategy for cavernous transformation of portal vein and basic research about its etiology is rarely reported. OBJECTIVE:To establish the models of cavernous transformation of portal vein, detect the expression of matrix metal oproteinase-2,-9 (MMP-2, MMP-9) and tissue inhibitors 1, 2 of metal oproteinase (TIMP-1, TIMP-2) in rat portal vein and peripheral tissue, and discuss the roles in the process of peripheral angiogenesis. METHODS:Eighty Sprague-Dawley rats were randomly divided into three groups. The rat models of cavernous transformation of portal vein were established with partial coarctation in portal vein by using 21 G blunt pinhead. Control group was normal rats without operation (samples were harvested after portal vein radiography). Model group and sham operation group were divided into three groups respectively according to different time points, namely 2, 4 and 6 weeks after operation. Rats of each group were randomly chosen at week 2, 4 and 6 after operation to observe the formation of col ateral circulation of portal vein and its peripheral tissues by performing portal vein radiography. CD31 was detected by immunohistochemistry. The expression of MMP-2, MMP-9, TIMP-1, TIMP-2 mRNA and protein in portal vein and peripheral tissue were determined by RT-PCR and immunohistochemistry respectively. RESULTS AND CONCLUSION:Peripheral angiogenesis of model group was increased obviously by portal vein radiography and immunohistochemistry. RT-PCR and immunohistochemistry results demonstrated that, compared with the control group and sham operation group, the expression of MMP-2 mRNA and protein in model group were significantly increased at weeks 2, 4, and 6 (P0.05). Ratio of MMP-2/TIMP-2 of model group was significantly higher than that of control group and sham operation group (P<0.05) at week 2. the rat models of cavernous transformation of portal vein have low mortality, high success rate and are stable. Upregulation of the expression of MMP-2, MMP-9 and the disbanlance of the ratio of MMP-2/TIMP-2 might contribute to the peripheral angiogenesis in rats with cavernous transformation of portal vein.
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BACKGROUND:It is presumed that urinary trypsin inhibitor could have protective effects on local and systemic tissues and could inhibit osteoclast proliferation and activation under long-term chronic inflammation conditions and in ischemic and anoxic environment which was induced by prosthetic wear. OBJECTIVE:To investigate the inhibitory effect of ulinastatin on receptor activator for nuclear factor-κb ligand-induced differentiation, proliferation and osteoclastogenesis of RAW264.7 cells and its effects on matrix metal oproteinase-2, matrix metal oproteinase-9 expression level and activity. METHODS:Mouse monocyte/macrophage cellline RAW264.7 was treated with different concentrations of urinary trypsin inhibitor (0, 500, 5 000 U/mL) for 24, 48 and 72 hours. Experiments were divided into four groups:the blank group (RAW264.7 cells), receptor activator for nuclear factor-κb ligand-induced group (0 U/mL ulinastatin), 500 U/mL ulinastatin group and 5 000 U/mL ulinastatin group. RESULTS AND CONCLUSION:(1) MTT results indicated that there was no significant difference on the proliferation of RAW264.7 cells treated with urinary trypsin inhibitor at 0-5 000 U/mL (P>0.05) (2) Tartrate-resistant acid phosphatase staining results revealed that compared with receptor activator for nuclear factor-κb ligand-induced group, the number of tartrate-resistant acid phosphatase-positive cells was significantly less in the ulinastatin group (P<0.05), showing a time-dose dependent manner. (3) Immunohistochemisical results found that compared with receptor activator for nuclear factor-κb ligand-induced group, the percentage of matrix metal oproteinase-9-positive cells was apparently lower in the ulinastatin group. (4) Western blot assay results demonstrated that matrix metal oproteinase-9 expression was low in the RAW264.7 cells alone. At 48 hours after addition of receptor activator for nuclear factor-κb ligand, matrix metal oproteinase-9 protein expression was large. At 72 hours after culture in the 5 000 U/mL ulinastatin group, matrix metal oproteinase-9 protein expression was evidently reduced. (5) Gelatin zymography results showed that compared with the receptor activator for nuclear factor-κb ligand-induced group, matrix metal oproteinase-9 expression was significantly lower in the 5 000 U/mL ulinastatin group (P<0.05). Results suggested that urinary trypsin inhibitor inhibited receptor activator for nuclear factor-κb ligand-induced osteoclastogenesis and diminished matrix metal oproteinase-9 expression and activity.
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BACKGROUND:Cel transplantation offers a new promise of rebuilding the damaged myocardium. But the results of them are not consistent. It is not clear if the transplanted cel s can permanently improve heart function and the mechanism underlying this therapeutic effect. OBJECTIVE:To study the effect of intracoronary autologous bone marrow mononuclear cel transplantation on cardiac function, and angiogenesis and cytokine production in canines with acute myocardial infarction. METHODS:Left anterior descending coronary artery ligation was used to produce acute myocardial infarction models in hybrid canines. Bone marrow mononuclear cel s were harvested by using puncture of anterior crest and posterior superior iliac spine to prepare cel suspension. Sixteen hybrid canines were randomly divided into transplantation group (n=10) and control group (n=6). Bone marrow mononuclear cel s (transplantation group, n=10) or normal saline (control group, n=6) were intracoronarily infused into infarction-related arteries 2 hours after acute myocardial infarction. To evaluate the heart function, we used echocardiography at 2 hours and 6 weeks after acute myocardial infarction. Capil ary density was assessed 6 weeks after transplantation by using von Wil ebrand factor test. The mRNA levels of vascular endothelial growth factor 188, vascular endothelial growth factor 164, basic fibroblast growth factor and matrix metal oproteinase-9 in the infarct area were determined by reverse transcription-PCR at 6 weeks after transplantation. RESULTS AND CONCLUSION:In contrast to the control group, ejection fraction and stroke volume at 6 weeks after transplantation increased significantly in the transplantation group. The transplantation group had a greater amount of new vessels in the peri-infarct area than the control group. Compared with the control group, the mRNA levels of vascular endothelial growth factor 188, vascular endothelial growth factor 164, and basic fibroblast growth factor significantly increased in the transplantation group, but the mRNA level of matrix metal oproteinase-9 significantly decreased in the transplantation group. These findings suggest that intracoronary transplantation of autologous bone marrow mononuclear cel s may improve the cardiac function, and increase capil ary density, especial y in the border zone of infarcted myocardium. Otherwise, bone marrow mononuclear cel transplantation can increase the mRNA levels of vascular endothelial growth factor 188, vascular endothelial growth factor 164, and basic fibroblast growth factor, but decrease the mRNA level of matrix metal oproteinase-9.