基质金属蛋白酶组织抑制剂-2基因转染对平滑肌细胞的影响

目的探讨基质金属蛋白酶组织抑制剂(TIMP)2基因对体外培养大鼠主动脉平滑肌细胞(SMCs)增殖和迁移能力的影响。方法利用重组腺病毒载体,将目的基因导入宿主细胞,建立转基因细胞株。应用逆转录聚合酶链反应(RTPCR)和酶联吸附试验(ELISA)检测TIMP2表达。同时应用细胞直接计数法和侵袭小室研究转染细胞体外增殖和侵袭情况。结果转染TIMP2基因的平滑肌细胞基因组中存在有590bp目的基因特异性片断,TIMP2蛋白在转染后平滑肌细胞中获得稳定表达和分泌。转染后平滑肌细胞生长受到明显抑制。AdhTIMP2组透过人工基底膜的细胞数为21.38±12.81,与正常对照组和AdCMV组(53.64±11.27,44.23±12.75)比较,差异有统计学意义(P<0.05)。结论重组腺病毒载体介导TIMP2基因转染对体外生长主动脉平滑肌细胞增殖和侵袭有明显抑制作用。     

基质金属蛋白酶及抑制因子与腹主动脉瘤临床病理特点的相关性研究

目的：研究基质金属酶（MMP）-2、MMP-9及抑制因子TIMP-1在腹主动脉瘤中的表达及与临床病理特征之间的关系。方法：应用免疫组化PV-9000通用型二步法对70例腹主动脉瘤和15例正常腹主动脉标本中的MMP-2、MMP-9及TIMP-1表达进行检测。结果：腹主动脉瘤组织中MMP-2和MMP-9蛋白表达阳性率明显高于正常腹主动脉组织,TIMP-1蛋白表达阳性率和正常腹主动脉没有统计学差异,（X^2=0.103,P=0.991）;MMP-2蛋白的表达与腹主动脉瘤的直径呈负相关（X^2=13.785,P=0.032）,MMP-9蛋白的表达与患者临床症状,腹主动脉瘤直径、破裂有相关性,（P〈0.05）,TIMP-1蛋白表达阳性率与临床病理特征无相关性（X^2=0.103,P=0.991）。结论：腹主动脉瘤组织中MMP高表达和TIMP的相对弱表达在腹主动脉瘤发生、发展过程中起重要作用,MMP-9可以预测腹主动脉瘤的自然病程从而作为腹主动脉瘤手术治疗的指征之一。     

血吸虫肝纤维化中基质金属蛋白酶及抑制剂的表达

目的观察基质金属蛋白酶(MMP)-13,基质金属蛋白酶组织抑制剂(TIMP)-3及α-平滑肌肌动蛋白(α-SMA)在晚期日本血吸虫病家兔肝脏和正常家兔肝脏中的表达。方法应用免疫组织化学和半定量Western免疫印迹方法检测了血吸虫病肝纤维化模型家兔42只及正常家兔8只中的MMP-13、TIMP-3、α-SMA的表达。结果正常对照组的MMP-13、TIMP-3、α-SMA的表达分别是(3.20±0.45)、(0.76±0.32)、(0.92±0.28)%;血吸虫病纤维化肝脏中的MMP-13、TIMP-3、α-SMA的表达分别是(10.56±3.62)、(6.50±2.56)、(5.78±2.98)%,较正常对照组明显增加(P<0.01),主要表达于成纤维细胞,肌成纤维细胞中,以纤维间隔及汇管区最明显,且三者来源定位基本一致。结论在血吸虫病肝纤维化中,成纤维细胞及肌成纤维细胞是MMP-13、TIMP- 3、α-SMA表达的主要细胞,TIMP-3的表达增高在肝纤维化的发生、发展中具有重要作用。     

Matrix metalloproteinase 9 and tissue inhibitor of metalloproteinase 1 in vesicoureteral reflux

The aim of this study was to investigate whether urine levels of matrix metalloproteinase 9 (uMMP9) and tissue inhibitor of metalloproteinase 1 (uTIMP1) are novel biomarkers of vesicoureteral reflux (VUR) and to determine the optimal cut-off levels of these enzymes to predict VUR in children. The study group consisted of 67 children with VUR and 20 healthy children. Urine MMP9 and TIMP1 levels were measured by an enzyme-linked immunosorbent assay. Children with VUR had significantly higher uMMP9 (1,539.8 vs. 256.4 pg/mL; p = 0.0001) and uTIMP1 (182 vs. 32.6 pg/mL; p = 0.0001) levels than healthy children. For the prediction of VUR, the sensitivity of uMMP9 was 67%, with a specificity of 85% [cut-off value 1,054 pg/mL; area under the curve (AUC) 0.77], and the sensitivity of uTIMP1 was 74%, with a specificity of 65% (cut-off value 18.7 pg/mL; AUC 0.73). Both uMMP9 and uTIMP1 levels were significantly higher in patients with renal scar (uMMP9: 3,117.3 vs. 1,234.15 pg/mL; p = 0.0001; uTIMP1: 551.05 vs. 128.64 pg/mL; p = 0.0001). Urine MMP9 levels had a sensitivity of 81.2%, with a specificity of 85% to predict renal scar in the VUR group (cut-off 1,054 pg/mL; AUC 0.88). The sensitivity of uTIMP1 was 75%, with a specificity of 90% to predict renal scar (cut-off 243.7 pg/mL; AUC 0.82). Based on these results, we suggest that uTIMP1 may be a useful marker to predict renal scarring with a different cut-off value from VUR and a high specificity at this cut-off point. Although uMMP9 seemingly cannot distinguish renal scar from VUR, the simultaneous increase in the level of both markers may indicate ongoing renal injury due to VUR.     

基质金属蛋白酶在腹主动脉瘤组织中的表达

目的探查基质金属蛋白酶类（MMPs）在腹主动脉瘤（AAA）组织中产生的源泉。方法采用间质胶原酶（MMP-1）和明胶酶以MMP-9）的mRNA探针在20例AAA组织及4例正常人腹主动脉组织的切片上行原位杂交实验结果MMP-1及MMP-9在巨噬细胞、平滑肌细胞和淋巴细胞均有表达,其中巨噬细胞的MMPs表达强烈。结论MMPs在AAA的形成和扩张中发挥重要作用,炎性细胞是产生MMPS的主要源泉,并影响问质细胞的MMPS表达。     

Clinical significance of tissue inhibitor of metalloproteinase and matrix metalloproteinase mRNA expression in thymoma

BACKGROUND: Matrix metalloproteinases (MMPs) are proteolytic enzymes which degrade extracellular matrix and basement membrane. There is much evidence that their increased expression is correlated with tumor aggressiveness in various carcinomas. Tissue inhibitor of metalloproteinases (TIMPs) are the specific inhibitors of MMPs. MMPs and TIMPs are considered to play an important role in carcinoma invasion and metastasis. We hypothesized that MMPs and TIMPs also play an important role in thymoma. MATERIALS AND METHODS: This study included 34 thymoma cases. The mRNA levels of MMP-1, -7, and -9, TIMP-1 and -2, and GAPDH were quantified by real-time polymerase chain reaction using LightCycler. We also performed immunohistochemistry for TIMP-1. RESULTS: The TIMP-1/GAPDH mRNA expression level was significantly higher in invasive (stage II-IV) thymomas (means +/- SE, 81.4 +/- 28.1) than in noninvasive (stage I) thymomas (30.9 +/- 8.3, P = 0.026). The MMP-1/GAPDH mRNA expression level was also higher in invasive thymomas (19.7 +/- 7.5) than in non invasive thymomas (2.26 +/- 1.72, P = 0.020). Immunopositivity of TIMP-1 was localized in stromal cells adjacent to the advancing margin of the tumor. CONCLUSIONS: These findings suggest that TIMPs and MMPs play an important role in the invasion of thymoma.     

基质金属蛋白酶-9及金属蛋白酶组织抑制剂-1对甲状腺乳头状癌诊断的意义

目的:探讨基质金属蛋白酶-9(MMP-9)和金属蛋白酶组织抑制剂-1(TIMP-1)在甲状腺肿瘤组织中的表达。方法:制备组织芯片,采用免疫组织化学和原位杂交技术检测56例甲状腺癌组织、56例癌旁组织和40例良性甲状腺病变组织中MMP-9和TIMP-1的表达情况。结果:MMP-9和TIMP-1蛋白在甲状腺癌组织的阳性表达率为71.4%和57.1%,MMP-9 mRNA和TIMP-1 mRNA在甲状腺癌组织中阳性表达率为67.9%和62.5%,均明显高于癌旁和良性甲状腺病变组织(P<0.05)。在甲状腺癌组织中,MMP-9、TIMP-1蛋白和MMP-9、TIMP-1 mRNA的表达,分别呈负相关性(RS=-0.309、-0.264,P<0.05)。结论:MMP-9和TIMP-1在组织中的检测有助于甲状腺癌的判断,并可作为预后评估的重要参考。     

基质金属蛋白酶抑制剂对胶质瘤细胞和胶质瘤干细胞体外侵袭抑制作用的比较

目的 观察基质金属蛋白酶(MMP)抑制剂对常规贴壁培养的人脑胶质瘤细胞和胶质瘤干细胞侵袭能力的影响及两者间的差异.方法 应用一种三维胶原凝胶侵袭模型,将常规贴壁培养的胶质瘤细胞与无血清、悬浮细胞球条件下培养的胶质瘤干细胞(1. 0×104～1.5×104个/样本)分别接种于该模型中,加入不同浓度的MMP抑制剂GM6001(0、25、50、75、100 μmol/L)培养4 d,通过观察各组细胞侵袭距离及其差异,判断该抑制剂对两种细胞侵袭能力的影响及差异.结果 两组细胞对GM6001的抑制作用表现出不同程度的剂量依赖性,75μmol/L GM6001对常规培养的胶质瘤细胞侵袭抑制最明显,而25 μmol/L GM6001对胶质瘤干细胞的侵袭抑制最显著.结论 MMP抑制剂在体外对胶质瘤细胞具有侵袭抑制作用,胶质瘤干细胞显示出对MMP抑制剂更高的敏感性.     

胃癌病人之血浆金属蛋白酶和组织抑制物水平及其临床意义

目的:探讨不同分期胃癌病人之血浆MMP-2、MMP-9及TIMP-1水平与病人个体预后之间的关系。方法:运用酶联免疫分析法(ELISA)测定不同分期的胃癌病人、良性胃病病人和健康志愿者血浆中MMPs/TIMP水平,逐例随访胃癌病人。结果:胃癌病人血浆MMP-2、MMP-9及TIMP-l的水平分别为(21.7±6.3)ng/ml(、17.9±9.2)ng/ml和(39.5±13.3)ng/ml;较良性胃病病人和健康志愿者显著升高,差异有显著性意义,尤其是Ⅳ期胃癌病人。胃癌病人血浆中MMP-2水平与肿瘤浸润深度、转移(包括淋巴结和远处)相关,T1MP-1与转移相关,MMP-2水平也与病人生存期相关。结论:血浆MMP-2可以作为预测胃癌病人预后的标志物。     

Presence of matrix metalloproteinases and tissue inhibitor of matrix metalloproteinase in human sperm

The matrix metalloproteinases (MMPs) are a family of proteolytic enzymes that degrade protein components of the extra-cellular matrix. The necessity of breakdown of physical barriers in the fertilization process suggests that MMPs, along with their tissue inhibitors (TIMPs), might be involved in this task. We have examined the presence of MMP and TIMP in normal and abnormal human sperm samples by gel zymography and Western blot analysis. Thirty-five normal sperm samples and 35 abnormal sperm samples were examined in this study. Gel zymography showed 92-, 72-, 62-, and 28-kd molecular-weight bands exhibiting gelatin-degrading activity in both normal and abnormal sperm samples. The 92-, 72-, and 62-kd bands with gelatinolytic activity are consistent with pro-MMP-9, pro-MMP-2, and active MMP-2, respectively (pro-MMP being the zymogen of MMP). Western blot analysis showed the presence of TIMP-1 in both normal and abnormal sperm samples. A higher 28-kd activity and a lower 92-kd MMP activity in normal sperm samples relative to abnormal samples were detected. No marked difference in TIMP-1, 72-kd, and 62-kd release was observed between normal and abnormal sperm samples. In conclusion, this is the first report of MMP activity in normal and abnormal human sperm samples and of TIMP presence in sperm samples. The data indicate a different MMP profile between normal and abnormal sperm samples, with a higher 28-kd activity and a lower 92-kd MMP activity in normal relative to abnormal samples.     

川崎病患儿基质金属蛋白酶-9及其组织抑制物-1血清水平变化的临床意义

目的通过对川崎病(KD)患儿血清基质金属蛋白酶(MMP)-9及其抑制物组织基质金属蛋白酶抑制物(TIMP)-1水平的测定,探讨MMP-9 及TIMP-1与川崎病发病及其冠状动脉并发症发生发展的关系.方法采用酶联免疫吸附检测(ELISA)法对30例KD无冠状动脉病变(CAL)及9例KD合并CAL患儿急性期和恢复期血清MMP-9、TIMP-1水平进行检测,并与15例其他发热性疾病患儿及18例正常健康儿童进行对照.结果急性期KD有CAL组和无CAL组血清MMP-9、TIMP-1、MMP-9/TIMP-1分别是896.2±81.7、342.6±43.1、2.5±0.6和284.3±40.9、389.5±20.8、0.8±0.2,均较发热对照组及正常对照组(87.9±18.9、251.5±13.0、0.3±0.1和24.6±2.8、90.0±4.2、0.3±0.02)明显增加(P<0.01),且有CAL组的MMP-9及MMP-9/TIMP-1较无CAL组增高更为显著(P<0.01);恢复期KD无CAL组患儿血清MMP-9、TIMP-1、MMP-9/TIMP-1降至正常水平(26.4±7.6、95.6±5.8、0.2±0.1),而有CAL组患儿虽较急性期亦明显下降(220.0±28.3、258.9±15.0、0.9±0.3),但仍然明显高于正常对照组(分别P<0.01和P<0.05),发热对照组血清MMP-9及TIMP-1水平亦较正常对照组明显增高(P<0.01),但其MMP-9/TIMP-1与正常对照组比较差异无显著性(P>0.05).结论 MMP-9及TIMP-1参与了川崎病的病理生理过程,血清MMP-9升高及MMP-9/TIMP-1持续失衡可能与川崎病冠状动脉炎及动脉瘤的形成有关.     

终末期肾脏病患者血管钙化与基质金属蛋白酶抑制剂

终末期肾脏患者中,普遍存在血管钙化。钙化不仅是钙盐沉积的过程,也是一个细胞介导的活跃过程。动脉粥样硬化的形成过程是细胞外基质（ECM）重建的过程,而基质金属蛋白酶（MMPs）及其抑制剂（TIMPs）在ECM的降解重建过程中有重要作用,提示MMP／TIMP系统在动脉粥样硬化中起重要作用。本文就ESRD患者血管钙化机制及MMP／TIMP与血管动脉粥样硬化方面的研究作一综述。     

直肠癌术前动脉插管化疗后手术时机的选择

探讨直肠癌患者术前动脉管化疗后的手术时机。方法对２７例Ｄｕｋｅｓ’Ｂ－Ｃ期直肠癌患者,选择肠系膜下动脉或左髂内动脉,以５－氟脲嘧啶６００ｍｇ／ｍ＾２、丝裂霉素１５ｍｇ／ｍ＾２、表阿霉素３０ｍｇ／ｍ＾２、丝裂霉素１５ｍｇ／