Purpose To identify matrix metalloproteinase (MMP) in human vitreous samples of diabetic vitreoretinopathy (DR) and other ocular diseases (non-DR) and to probe the related factors of MMP expression． Methods Thirty-one diabetic and 17 non-diabetic vitreous samples (nine macular hole and eight epiretinal membrane patients) were examined． Samples were concentrated and subjected to substrate zymography to conduct a quantitative analysis of MMP-2,9 activity． The technology of Western blotting against anti-human MMP-2,9 was performed to identify MMP in vitreous samples． Results Vitreous samples both from DR patients and from non-DR patients showed a single band at the position of 72 kDa, correspondin g to MMP-2． Quantitative analysis revealed that diabetic vitreous showed higher MMP-2 activity than non-DR, although the difference was not significant.45.2% of DR patients showed MMP-9, but no expression in non-DR．Among DR samples, the positive ratio of MMP-9 in partial posterior vitreous detachment (PVD)(66．7%) was significantly higher than that of complete PVD (15．4%)． Western blotting study confirmed the expression of MMP-2 and MMP-9． Conclusion There is no obvious difference of MMP-2 activity between DR and non-DR． MMP-9 may be involved in the pathogenesis of diabetic vitreor etinopathy and the deterioration of proliferative change． (Chin J Ocul Fundus Dis, 2001,17:195-197
Abstract: Objective To observe the expression of integrinlinked kinase (ILK) and matrix metalloproteinases9 (MMP9) in human nonsmall cell lung cancer (NSCLC) and investigate the correlation of ILK and MMP9 expression with the prognosis of NSCLC. Methods The expression of ILK and MMP9 in 75 specimens of NSCLC resected from January 2002 to January 2004 were detected by immunohistochemistry. According to the median of integral optical density (IOD), all patients were divided into the high or low ILK expression group and the high or low MMP-9 expression group. The relativity of ILK and MMP9 was determined, and the relationship of survival time with clinical features including expression of ILK and MMP-9 was compared by Logrank test. Results Both ILK and MMP-9 were expressed in NSCLC specimens. The expression between ILK and MMP-9 was positively correlated in 75 patients of our group (r=0.79, Plt;0.05). Patients with lower expression of ILK and MMP9 had a significantly longer survival time than those with higher expression of ILK and MMP-9 in the postoperative followup (χ2=15.067,14301,Plt;0.05). The survival time was not correlated with sex,age,smoking history or pathological type(χ2=0450,0078, 1.460, 1.623，Pgt;0.05), while tumor diameter, lymph node metastasis, TNM stage, the expression of ILK and MMP-9 significantly influenced the survival time (χ2=3.963, 15.169,20.529, 15.067，14.301，Plt;0.05). Conclusion The expression of ILK and MMP9 affects the prognosis of NSCLC. MMP-9 may advance infiltration and metastasis of tumor cells through ILK pathway. In summary, the expression of ILK and MMP9 may play an important role in the evaluation of prognosis for patients with NSCLC.
Objective To investigate the expression of matrix metallo proteinase (MMP)-2 and MMP-9 in rats′optical nerves after extrusion wound. Methods We set up the model of rats with extrusion wound of the optical nerves, detected activity changes of MMP-2 and MMP-9 in the optical nerves by gelatin zymography, identified the attribute by Western blotting, and verified the expression of mRNA of MMP-2 and MMP-9 by reverse transcriptase-polymerase chain reaction (RT-PCR ). Results MMP-2 existed in normal optial nerves and optical nerves with extrusion wound, while MMP-9 was only detected in the latter. The expression of MMP-9 was the highest 1 day after the extrusion wound, while that of MMP-2 was the highest 7 days after the extrusion wound. Conclusions MMP-2 and MMP-9 may participate in the pathological recovery process of optical nerves after extrusion wound. The glial cells in the optical nerves may be one of the sources of MMP-2 and MMP-9. (Chin J Ocul Fundus Dis，2003，19：269-332)
Objective To investigate the feasibil ity of alendronate (ALN) in treating osteoarthritis (OA) by observing the effects of ALN on interleukin 1β (IL-1β) induced chondrocytes of rat in vitro. Methods The chondrocytes of knee articular surface from 15 SD rats (1-month-old, female or male, weighing 100-150 g) were cultured. The chondrocytes were observed by inverted phase contrast microscope and identified by toluidine blue staining and HE staining. The third passage chondrocytes were divided into 3 groups: the chondrocytes were cultured with DMEM for 5 days in group A, with 10 ng/mL IL-1β for 2 days and with DMEM for 3 days in group B, and with 10 ng/mL IL-1β for 2 days and with 1 × 10-6 mol/L ALN for 3 days in group C. Immunocytochemistry and real-time PCR were performed to determine the expression levels of collagen type II (Col II), matrix metalloproteinase 13 (MMP-13), and β-catenin. Results Toluidine blue staining proved that the cultured cells were chondrocytes. The integrated absorbency (IA) value of Col II in group C (10.290 7 ± 0.499 2) was lower than that in group A (15.377 0 ± 0.571 8) and higher than that in group B (5.463 2 ± 0.450 4), showing significant differences (P lt; 0.05). The IA value of MMP-13 in group C (3.068 6 ± 0.205 6) was significantly lower than that in group B (6.998 1 ± 0.329 7, P lt; 0.05), but there was no significant differenc when compared with group A (2.777 5 ± 0.199 6, P gt; 0.05). The IA value of β-catenin in group C (6.611 7 ± 0.381 8) was lower than that in group B (11.799 9 ± 0.348 7) and higher than that in group A (4.390 3 ± 0.551 9), showing significant differences (P lt; 0.05). The mRNA expression of Col II in group C was significantly higher than those in groups A and B (P lt; 0.05), the mRNA expression of MMP-13 in group C was significantly lower than that in group B (P lt; 0.05) but there was no significant difference when compared with group A (P gt; 0.05). The mRNA expression of β-catenin in group C was significantly lower than that in group B (P lt; 0.05) and higher than that in group A (P lt; 0.05). Conclusion ALN can protect rat chondrocyte from OA induced by IL-1β in vitro possibly by upregulating Col II and inhibiting the expression of MMP-13 and β-catenin in the chondrocytes.
ObjectiveTo explore the inhibition action of valproic acid to inflammatory cells and smooth muscle cells then to find out that valproic acid (VPA) can repress rat thoracic aortic aneurysm or not. MethodsThe model of rat thoracic aortic aneurysm was built through the method of soaking the adventitia of artery using porcine pancreatic elastase (PPE). The rats were divided into three groups:a normal saline blank control group (a C group), an adventitia soaked PPE group (a P group), and adventitia soaked PPE plus intraperitoneal injection by injecting intraperitioneal VPA 200 mg/kg for seven days (a PV group).The animals of the three groups were all using vascular ultrasound to detect blood vessel diameter. Animals were killed after operation to observe the general morphology of vascular aneuysm and do the immunohistochemial, morphological, protein analysis of interleukin 1 (IL-1), interleukin 6 (IL-6), smooth muscle 22 alpha (SM22α), matrix metallopeptidase 2 (MMP-2), MMP-9 and Western blot by drawing animals on the 14th day. ResultsThe vessels diameter in the PV group was narrower than that in the P group (P value<0.05). HE staining, immunohistochemistry and Western blot displayed that the cells in the P group were in disorder arrangement and interstitial disorder while the cells in the PV group maintained better albumin layer. The protein expressions of IL-1, IL-6, MMP-2, and MMP-9 in the PV group decreased except that SM22α increased. ConclusionVPA can inhibit phenothpic transforming of aneurysm inflammatory cells and smooth muscle cells, reduce the levels of cell proliferation, decrease the secretion of matrix metalloproteinases, and depress tumor growth of rat thoracic aorta.
Objective Astragalus polysaccharide (APS) has promoting angiogenesis function. To explore the effects of APS collagen sponge on enhancing angiogenesis and collagen synthesis so as to provide evidence for the future tissue engineering appl ication as a kind of angiogenic scaffold. Methods APS collagen sponges were prepared by covalent binding with collagen polypeptides by using of crossl inking agents at the ratio of 1 ∶ 1 (W/W). Twenty 10-week-old SpragueDawley rats (10 males and 10 females, and weighing 200-250 g) were selected. Longitudinal incision was made at both sides of the back to form subcutaneous pockets. APS collagen sponges of 5 mm × 5 mm × 5 mm at size were implanted into the left pockets as the experimental group, collagen sponges without APS of the same size into the right pockets as the control group. The general conditions were observed after operation. At 3, 7, 14，and 21 days, 5 rats were sacrificed and the samples were harvested to count the number of microvessels, to measure the contents of the hydroxyprol ine (Hyp), and to detect the mRNA expressions of angiopoetin 1 (Ang1), matrix metalloproteinases 9 (MMP-9), and tissue inhibitors of metalloproteinases 1 (TIMP-1). Results All rats were al ive during experiment period. The number of microvessels increased gradually, and reached the peak at 14 days in 2 groups; the expermental group was significantly higher than the control group (P lt; 0.05). The contents of Hyp increased gradually in 2 groups, and the experimental group was significantly higher than the control group (P lt; 0.05). The mRNA expressions of Ang1 and MMP-9 in the experimental group were significantly higher than those in the control group at 3, 7, and 14 days (P lt; 0.05); the mRNA expression of TIMP-1 in the experimental group was significantly lower than that in the control group at 3 days and was significantly higher at 14 and 21 days (P lt; 0.05). Conclusion The APS collagen sponges can improve angiogenesis and collagen synthesis in wound heal ing by regulating the expressions of Ang1, MMP-9, and TIMP-1.
ObjectiveTo summarize the biological function of extracellular matrix metalloproteinase inducer (EMMPRIN) in tumor progression, and its roles in clinical diagnosis and treatment in recent years. MethodsLiteratures about the recent studies on molecular structure of EMMPRIN and biological function in tumor progression were reviewed according to the results searched from PubMed database. ResultsEMMPRIN play important roles in the tumor progression, involved in inducing the degradation of extracellula matrix, promoting angiogenesis, inhibiting apoptosis, enhancing chemoresistance and so on. ConclusionEMMPRIN could be a potential therapeutic target in turmor.
Objective To observe the effect of epidermal growth factor (EGF) on the proliferation, adhesion, invasiveness and the activation of nuclear factor-κB (NF-κB), matrix metalloproteinases (MMPs) expression and explore related mechanisms in pancreatic cancer cells. Methods Cell invasion assay, proliferation assay and adhesion assay were used to examine the proliferation, adhesion and invasiveness of pancreatic cancer cells, respectively. NF-κB activity was detected by electrophoretic mobility shift assay (EMSA), and MMPs protein and mRNA expressions were investigated by gelatin zymography, Western blot and reverse transcriptase-polymerase chain reaction (RT-PCR). Results EGF increased the invasiveness of pancreatic cancer cell in a dose-dependent manner (P＜0.05), but did not affect cell proliferation or adhesion. The expressions of MMP-9 mRNA and protein significantly increased after induction by EGF and were highest when EGF concentration was 50 ng/ml, while there was no effect on the expressions of MMP-2 mRNA and protein. Furthermore, NF-κB activity increased with increased concentration of EGF in a concentration-dependent manner (P＜0.05). In addition, NF-κB activity and the expressions of MMP-9 mRNA and protein by pretreatment with both pyrrolidine dithiocarbamate (PDTC) and EGF decreased when compared that by pretreatment with EGF alone. The invasiveness of pancreatic cancer cell by pretreatment with both PDTC and EGF decreased when compared that by pretreatment with EGF alone and nothing (P＜0.05).Conclusion The findings indicate that the NF-κB-mediated MMP-9 induction is essential for EGF-induced invasiveness in pancreatic cancer cells, which can be inhibited by PDTC.
ObjectiveTo investigate the expression of amniotic fluid levels and blood serum levels of matrix metalloproteinases-8 (MMP-8) and interleukin-6 (IL-6) in women with preterm delivery. MethodsBetween January 2010 and December 2012, we collected the amniotic fluid of 102 preterm pregnant women and 98 full term pregnant women and analyzed the MMP-8 levels and IL-6 levels in amniotic fluid and blood serum. Meanwhile, we also collected the amniotic fluid to do bacterial culture. ResultsThe amniotic fluid levels of MMP-8 in preterm pregnant women were higher than those in full term pregnant women [(320.45±59.88) vs (153.72±29.12) ng/mL, P<0.05], but there was no obvious discrepancy in the blood serum levels of MMP-8 in the two groups [(9.56±2.11) vs (9.42±2.01) ng/mL, P>0.05]. Both amniotic fluid levels and blood serum levels of IL-6 in preterm pregnant women were significantly higher than the full term pregnant women [(90.5±16.3] vs (20.6±12.5) μg/L, P<0.05; (159.2±20.4) vs (22.3±11.8) μg/L, P<0.05]. The positive bacterial culture rate of preterm pregnant women was higher than the full term pregnant women (8.8% vs 1.0%, P<0.05). ConclusionInfection is the most important reason for preterm pregnancy. MMP-8 level increases in the amniotic fluid, and the level of IL-6 in amniotic fluid and blood serum is a valuable clinical index for identifying premature delivery.
Objective To study the expressions and clinical significance of matrix metalloproteinases-2 (MMP-2) and vascular endothelial growth factor-C (VEGF-C) in patients with papillary thyroid carcinoma (PTC). Methods SP immunohistochemical technique was used to detect the expressions of MMP-2 and VEGF-C in 78 cases of PTC and 18 cases of thyroid benign tumors.Results The positive expression rates of MMP-2 and VEGF-C in PTC （80.77%, 75.64%） were significantly higher than those of the thyroid benign tumor （11.11%, 22.22%）, P＜0.05. The expressions of MMP-2 and VEGF-C were correlated to the degree of infiltration and lymph node metastasis in PTC: In those which infiltrated to or over the thyroid capsular, or had clinical neck lymph node metastasis, the positive expression rates were significantly higher than those in the other cases which had confined invasion of thyroid capsular or non-clinical metastasis of neck lymph node (P＜0.05). And during the follow-up of 41 patients who didn’t have clinical neck lymph node metastasis before operation, the positive expression rates of those who had clinical neck lymph node metastasis were significantly higher than those in the other patients who didn’t have neck lymph node metastasis (P＜0.05). There was significantly positive correlation between the expressions of MMP-2 and VEGF-C in PTC （Gamma=0.846， P＜0.05）. Conclusions MMP-2 and VEGF-C may be used to distinguish malignant and benign thyroid tumor; The expressions of MMP-2 and VEGF-C are correlated with the degree of infiltration and neck lymph node metastasis in PTC; Combined detection of MMP-2 and VEGF-C will be more accurate to predict condition of lymph node metastasis.