TIMP-1、MMP-2、MMP-9在大肠癌组织中表达的临床意义

目的 研究TIMP-1、MMP-2和MMP-9在大肠癌组织中表达的临床意义.方法 采用免疫组化SP法检测50例大肠癌、10例正常大肠黏膜组织中TIMP-1、MMP-2和MMP-9的表达.结果 TIMP-1、MMP-2、MMP-9在正常组织中的表达均较低,而它们在大肠癌组织中阳性率均较高(P<0.05).结论 TIMP-1、MMP-2、MMP-9与大肠癌临床病理参数有关系.TIMP-1、MMP-2和MMP-9可能是大肠癌侵袭转移的分子标记物.     

基质金属蛋白酶-9和组织金属蛋白酶抑制剂-1在食管鳞癌中的表达

目的探讨基质金属蛋白酶-9（MMP-9）和组织金属蛋白酶抑制剂-1（TIMP-1）在食管鳞癌中的表达及其临床意义。方法用免疫组化和Western blot法分别检测41例食管鳞癌患者的癌及相应正常组织中MMP-9和TIMP-1的表达变化。结果食管鳞癌组织中MMP-9阳性表达率与食管癌淋巴结及静脉转移有关；MMP-9的阳性表达率与表达量均显著高于TIMP-1；MMP-9和TIMP-1的表达呈负相关。结论MMP-9与食管鳞癌的侵袭转移有关，其机制可能与食管鳞癌组织中的MMP-9／TIMP-1平衡失调有关；MMP-9与TIMP-1联合检测有助于食管鳞癌生物学行为的判断。     

恶性肿瘤外周血基质金属蛋白酶2及其抑制物表达的临床研究

目的：研究基质金属蛋白酶2（MMP-2）及金属蛋白酶抑制剂-2（TIMP-2）表达在胃癌、肠癌转移及预后判断中的作用。方法：采用逆转录荧光实时定量PCR的方法检测58例结肠癌患者,52例胃癌患者外周血中MMP-2和TIMP-2 mRNA的表达水平。结果：胃癌、肠癌患者MMP-2表达均高于正常对照（P〈0．01）;其中,有局部浸润、淋巴结肿大、远处脏器转移者的MMP-2 mRNA和TIMP-2／MMP-2比值与未转移者差异有统计学意义（P〈0．01）;TIMP-2／MMP-2比值对判断胃癌、肠癌瘤转移的灵敏度分别为86．6％、89．2％,特异性为81．8％、83．3％。结论：基质金属蛋白酶2的表达升高与恶性肿瘤的转移密切相关,TIMP-2／MMP-2比值是恶性肿瘤转移和预后判断的良好指标。     

Disordered beta-catenin expression and E-cadherin/CDH1 promoter methylation in gastric carcinoma

AIM: To investigate the distribution of beta-catenin in nuclei or membrane/cytoplasm of gastric carcinoma cells, the relationship between E-cadherin gene methylation and its expression, and the role of beta-catenin and E-cadherin as potential molecular markers in predicting tumor infiltration. METHODS: Twenty-nine cases of gastric carcinoma, classified as diffuse and intestinal variants, were selected for study. Nuclear and cytoplasmic proteins were purified and beta-catenin content was detected by ELISA. DNA methylation of E-cadherin/CDH1 gene promoter was studied by methylation-specific PCR and compaired with E-cadherin expression detected by immunohistochemistry. RESULTS: In 27 cases of gastric carcinoma, the ratio of beta-catenin content between nuclei and membrane/cytoplasm was correlated with the T-classification (r = 0.392, P = 0.043). The significance was present between T2 and T3 groups. No correlation was detected between diffuse and intestinal variants in terms of their beta-catenin distribution. In 21 cases of diffuse variants of gastric carcinoma, there was a difference in E-cadherin expression between CDH1 gene-methylated group and non-methylated group (29 % vs 71 %, P = 0.027). No correlation between CDH1 gene methylation and T-classification was found, neither was the significance between E-cadherin expression and tumor infiltration grade. CONCLUSION: Comparative analysis of nuclear and membrane/cytoplasmic beta-catenin can predict local tumor infiltration. E-cadherin/CDH1 gene methylation is an important cause for its gene silence in diffuse variant gastric carcinoma. Methylation of CDH1 gene in the absence of E-cadherin is an early event in gastric carcinogenesis.     

Disordered beta-catenin expression and E-cadherin/CDH1 promoter methyiation in gastric carcinoma

AIM: To investigate the distribution of beta-catenin in nuclei or membrane/cytoplasm of gastric carcinoma cells, the relationship between E-cadherin gene methylation and its expression, and the role of beta-catenin and E-cadherin as potential molecular markers in predicting tumor infiltration. METHODS: Twenty-nine cases of gastric carcinoma, classified as diffuse and intestinal variants, were selected for study. Nuclear and cytoplasmic proteins were purified and beta-catenin content was detected by ELISA. DNA methylation of E-cadherin/CDH1 gene promoter was studied by methylation-specific PCR and compaired with E-cadherin expression detected by immunohistochemistry. RESULTS: In 27 cases of gastric carcinoma, the ratio of beta-catenin content between nuclei and membrane/ cytoplasm was correlated with the T-classification (r = 0.392, P = 0.043). The significance was present between T2 and T3 groups. No correlation was detected between diffuse and intestinal variants in terms of their beta-catenin distribution. In 21 cases of diffuse variants of gastric carcinoma, there was a difference in E-cadherin expression between CDH1 gene-methylated group and non-methylated group (29 % vs 71 %, P = 0.027). No correlation between CDH1 gene methylation and T-classification was found, neither was the significance between E-cadherin expression and tumor infiltration grade. CONCLUSION: Comparative analysis of nuclear and membrane/cytoplasmic beta-catenin can predict local tumor infiltration. E-cadherin/CDH1 gene methylation is an important cause for its gene silence in diffuse variant gastric carcinoma. Methylation of CDHl gene in the absence of E-cadherin is an early event in gastric carcinogenesis.     

MMP-9/TIMP-1表达失衡在子宫内膜异位症中的临床意义

目的探讨基质金属蛋白酶9(MMP9)及其抑制剂(TIMP1)在子宫内膜异位症(EMs)发生发展过程中的作用。方法收集50例EMs患者的异位内膜组织标本和20例在位内膜组织标本,采用免疫组化方法分别检测MMP9、TIMP1在两组组织中的表达。结果MMP9、TIMP1在异位内膜中多呈阳性或强阳性表达,但两者表达程度不同,前者随美国生育协会(rAFS)分期增高表达上调,后者则随rAFS分期增高表达下调,两者相比较差异具有显著性意义(P<005)。正常对照组的在位内膜中多呈弱阳性表达或无表达,两组比较差异极具显著意义(P<001)。结论MMP9/TIMP1表达失衡在EMs发生发展过程中可能有着重要作用。     

胃癌组织MMP-9基因与血清中蛋白相关性研究

目的了解胃癌组织中基质金属蛋白酶-9（MMP-9）基因表达与术前血清中蛋白水平关系，探讨MMP-9表达与胃癌临床病理参数之间相关性。方法45例胃癌组织通过半定量逆转录聚合酶链反应（semi-RT-PCR）、免疫组织化学染色（IHC）、酶联免疫吸附（ELISA）方法检测组织中mRNA及蛋白表达情况，测定术前血清中蛋白水平。结果肿瘤组织基质金属蛋白酶-9mRNA表达程度、蛋白染色阳性率、血清中蛋白水平均显著高于正常对照组（P〈0．01）；胃癌组织蛋白表达阳性率与肿瘤是否侵犯至浆膜层相关（P〈0．05）；术前血清中蛋白水平与肿瘤TNM分期、淋巴结转移显著相关（P〈0．01）。结论检测基质金属蛋白酶-9表达对胃癌有一定诊断意义；胃癌组织中蛋白表达率及术前血清中蛋白水平对临床肿瘤病情评估具有指导意义；也许多种机制参与了胃癌术前血清中MMP-9蛋白的分泌。     

Clinicopathological correlations of TIMP-1 and TIMP-2 in Hodgkin's lymphoma

OBJECTIVES: The influence of matrix-tumour interactions in Hodgkin's lymphoma is poorly characterised, although a large part of the tumour often consists of reactive tissue. The aim of the present study was to assess the clinicopathological role of two main inhibitors of matrix metalloproteinases, TIMP-1 and TIMP-2, in Hodgkin's lymphoma. MATERIALS AND METHODS: The TIMP-1 and TIMP-2 protein expressions were studied from paraffin-embedded tumour sections of 68 patients with Hodgkin's lymphoma by using immunostaining with TIMP-1 and TIMP-2-specific antibodies. The results of the stainings were compared with the clinicopathological disease characteristics. RESULTS: A total of 33.3% of the tumour tissue sections expressed TIMP-1 and 46.8% expressed TIMP-2. The expression of the TIMP-1 protein was found to be strongly associated with the nodular sclerosis subtype (P = 0.015) and the existence of a bulky tumour (P = 0.004) in Hodgkin's lymphoma. The expression of the TIMP-2 protein, on the other hand, correlated with the occurrence of B symptoms (P = 0.032). CONCLUSIONS: These results provide the first clinical evidence suggesting that TIMP-1 could promote growth of Hodgkin's lymphoma, and may be linked to connective tissue turnover in the nodular sclerosis subtype. However, TIMP-2 is shown to correlate with systemic symptoms.     

基质金属蛋白酶-9及其抑制物TIMP-1表达与醋柳黄酮治疗大鼠缺血-再灌注损伤心肌的关系研究

目的：观察基质金属蛋白酶（MMP）-9及其抑制物TIMP-1表达与醋柳黄酮治疗大鼠缺血-再灌注损伤心肌的关系研究,探讨醋柳黄酮对心肌缺血再灌注损伤的保护机制。方法：将50只大鼠随机分组：模型组、氯沙坦组、醋杉P黄酮大小剂量组及假手术组。结扎左冠状动脉前降支,30min后松开制作心肌缺血再灌注模型,免疫组织化学检测再灌注心肌组织MMP-9及其抑制物TIMP-1表达。结果：免疫组化结果示模型组较假手术组MMP-9、TIMP-1表达增高;醋柳黄酮大小剂量组较模型组MMP-9表达减少,TIMP-1表达增多。且有统计学意义。结论：醋柳黄酮对再灌注损伤心肌有保护作用,其作用机制与抑制心肌中MMP-9表达,促进TIMP-1表达有关。     

糖皮质激素对轻度持续性哮喘患者痰液中MMP-9/TIMP-1的影响

目的 观察糖皮质激素对轻度持续性哮喘患者痰液中MMP-9/TIMP-1的影响,探讨糖皮质激素降低气道重塑的机制.方法 轻度持续性支气管哮喘患者23例,采用酶联免疫吸附法(ELISA)测定使用吸入糖皮质激素前后痰液中MMP-9、TIMP-1、MMP-9/TIMP-1水平的变化并与肺功能指标进行相关性分析.结果 使用吸入糖皮质激素布地奈德后哮喘患者痰液MMP-9、MMP-9/TIMP-1水平显著降低(P<0.05).PEF、FEV1占预计值百分比与MMP-9/TIMP-1比率成负相关性(r=-0.402,r=-0.364,P<0.05).结论 糖皮质激素降低气道重塑的机制可能与其纠正MMP-9/TIMP-1失衡有关.     

Expression of matrix metalloproteinases (MMP-2 and -9) and tissue inhibitors of metalloproteinases (TIMP-1 and -2) in acute myelogenous leukaemia blasts: comparison with normal bone marrow cells

We compared the expression of matrix metalloproteinases (MMP-2 and MMP-9) and tissue inhibitors of metalloproteinases (TIMP-1 and TIMP-2) in bone marrow acute myelogenous leukaemia (AML) blasts and leukaemic cell lines (HEL, HL-60, K-562 and KG-1) with their expression in normal bone marrow cells. All AML samples and leukaemic cell lines tested expressed MMP-9 and/or MMP-2 mRNA and, accordingly, these gelatinases were secreted into media. Moreover, TIMP-1 and TIMP-2 mRNA and secreted proteins were demonstrated in all the AML samples. Although all the leukaemic cell lines expressed TIMP-1, the HL-60 cells also expressed TIMP-2. In contrast, normal steady-state bone marrow immature progenitor cells (CD34+ cells) did not express or secrete either MMP-2 or MMP-9, but more mature mononuclear cells from normal bone marrow expressed and secreted MMP-9. Also, normal bone marrow CD34+ cells and mononuclear cells expressed TIMP-1 and TIMP-2 mRNA, but these proteins were not detectable by reverse zymography. Furthermore, whereas bone marrow fibroblasts and endothelial cells secreted only latent MMP-2, the activated form of this enzyme was found in media conditioned by cells obtained from long-term cultures of normal and AML bone marrow adherent layers. Our finding of up-regulated production of gelatinases, TIMP-1 and TIMP-2 by leukaemic cells suggests that these proteins may be implicated in the invasive phenotype of AML.     

葶苈生脉饮对压力负荷大鼠心肌组织MMP-9及TIMP-1表达的影响

治疗CHF的作用机制之一.     

金属基质蛋白酶9及其组织抑制物1在肾组织中的表达和调控

目的:观察STZ鼠肾组织及高糖状态下正常人类系膜细胞(NHMC)中金属基质蛋白酶9(MMP-9)、金属基质蛋白酶组织抑制物1(TIMP-1)表达状况及磷酸肌酸激酶(PKC)抑制剂对其表达的影响,探讨糖尿病肾病(DN)时PKC抑制剂在细胞外基质降解中的作用。方法:Wistar大鼠随机分为正常对照组、DN模型组、PKC抑制剂组。PKC抑制剂组采用根皮素10mg/(kg.d)混悬液灌胃进行干预。第8周处死大鼠(每组6只)。检测24h尿蛋白定量、血肌酐水平。用免疫组化方法检测肾脏组织MMP-9、TIMP-1的表达。用ELISA方法检测肾脏组织PKC活性。体外实验,将NHMC置37℃,5%CO2培养箱中进行培养。并将NHMC分为N组(对照组):糖浓度5mmol/L,H组(高糖组):糖浓度30mmol/L,P组(高糖加PKC抑制剂):糖浓度30mmol/L加chelery thrine chloride 10-5mmol/L,M组(甘露醇组):甘露醇30mmol/L。于培养24、48、72h后用MTT法测定细胞增值。采用ELISA方法检测四组PKC活性。分别用RT-PCR及Western Blot检测各组MMP-9、TIMP-1mRNA及蛋白表达。结果:DN模型组尿蛋白排泄明显增加(P<0.01),血肌酐上升(P<0.05),PKC活性明显增高,MMP-9和TIMP-1出现表达,MMP-9/TIMP-1比值降低。PKC抑制剂干预后其尿蛋白排泄明显减少,血肌酐水平降低,PKC活性下降,MMP-9、TIMP-1表达上调,其MMP-9/TIMP-1比值增高。体外实验中,高糖能促进NHMC增殖,且NHMC的增殖状况随时间的递增而明显增加。高糖(30mmol/L)能增加系膜细胞PKC的活性,MMP-9、TIMP-1较高表达,MMP-9/TIMP-1比值降低。而PKC抑制剂使PKC的活性降低同时,MMP-9、TIMP-1表达上调,MMP-9/TIMP-1比值增高。结论:高糖可诱导PKC活性,PKC抑制剂能使MMP-9、TIMP-1表达上调,MMP-9/TIMP-1表达比值升高,推测PKC的活性状况可影响DN细胞外基质降解过程。     

MMP-9、TIMP-1在子宫内膜异位症患者子宫内膜中的表达及意义

目的观察基质金属蛋白酶-9（MMP-9）和基质金属蛋白酶组织抑制剂-1（TIMP-1）在子宫内膜异位症（EMs）患者子宫内膜中的表达,并探讨其在EMs发病机制中的作用.方法利用免疫组化法检测MMP-9和TIMP-1在35例EMs患者（观察组,其中未孕16例、已孕19例）异位内膜及在位内膜组织和35例正常人（对照组）子宫内膜中的表达.结果①MMP-9在观察组异位内膜中的表达高于在位内膜和对照组（P均＜0.05）,在位内膜中的表达高于对照组（P均＜0.05）;②TIMP-1、MMP-9/TIMP-1在观察组异位内膜中的表达低于在位内膜和对照组（P均＜0.05）;观察组未孕与已孕患者MMP-9和TIMP-1表达以及MMP-9/TIMP-1均无统计学差异（P均＞0.05）.结论MMP-9高表达以及MMP-9/TIMP-1的比例失调,使子宫内膜更具侵袭性,在EMs的发生、发展中起重要作用.     

E-cadherin and beta-catenin expression in Epstein-Barr virus-associated gastric carcinoma and their prognostic significance

AIM： To examine the role of E-cadherin and betacatenin in carcinogenesis and to assess their prognostic implication in Epstein-Barr virus-associated gastric carcinomas （EBV-GCs）. METHODS： We compared the frequency of E-cadherin and beta-catenin expression in 59 EBV-GCs and 120 non-EBV-GCs, and examined the association between patients＇ prognosis and the expressions of these proteins.
RESULTS： Neither the cellular-membranous nor the cytoplasmic E-cadherin expression showed any difference between EBV-GCs and non-EBV-GCs. On the other hand, loss of membranous expression of beta- catenin occurred more frequently in non-EBV-GCs than EBV-GCs [odds ratio = 0.41; 950 confidence interval （CI）, 0.19-0.90]. Furthermore, the nuclear and/or cytoplosmic expression of beta-catenin was seen more frequently in EBV-GCs than non-EBV-GCs （odds ratio = 2.23; 95% CI, 0.97-5.09）, and was observed in a larger proportion of carcinoma cells of EBV-GCs than non-EBV-GCs （P = 0.024）. Survival analysis for non-EBV-GC revealed that lymph node metastasis was significantly associated with poor prognosis （P 〈 0.001）. Among EBV- GCs, the depth of invasion （P = 0.005）, lymph node metastasis （P = 0.004） and an intestinal type by Lauren classification （hazard ratio = 9.47; 95% CI, 2.67-33.6） were significantly associated with poor prognosis. On the other hand, nuclear and/or cytoplasmic expression of beta-catenin was associated with a better prognosis in patients with EBV-GC （hazard ratio = 0.32; 95% CI, 0.11-0.93）. CONCLUSION： We observed more frequent preservation of beta-catenin in cell membrane and accumulation in nuclei and/or cytoplasm in EBV-GCs than in non-EBV- GCs. Factors involved in the prognosis of EBV-GCs and non-EBV-GCs are different in the two conditions.     

基质金属蛋白酶-9及其抑制剂在COPD合并肺间质纤维化大鼠中的作用

目的 观察基质金属蛋白酶-9(MMP-9)及组织型蛋白酶抑制剂-1(TIMP-1)在慢阻肺继发肺间质纤维化(PIFCOPD)大鼠肺组织中的变化,评价其与PIF-COPD的病理变化及发病机制的相关性.方法 雄性大鼠24只,随机分为正常对照组(1组),21天(2组)和42天(3组)模型组.以烟熏联合气管内滴注脂多糖建立PIF-COPD模型.免疫组化法检测肺组织中MMP-9及TIMP-1水平.结果 MMP-9和TIMP-1在1组有少量表达;在2和3组MMP-9为0.1970±0.0017和0.2414 ±0.0017;TIMP-1为0.1265 ±0.0018和0.2171 ±0.0023(与l组相比P＜0.05)水平逐渐增高且与肺间质纤维化的严重程度成正相关.结论 MMP-9及TIMP-1的水平与PIF-COPD的发病机制及病程有关.     

咳喘宁对支气管哮喘大鼠气道重塑及MMP-9、TIMP-1的影响

目的 观察咳喘宁对支气管哮喘大鼠气道形态学和基质金属蛋白酶-9（MMP-9）、基质金属蛋白酶组织抑制剂-1（TIMP-1）的影响.方法 40只SD大鼠随机分为正常组、模型组、咳喘宁高、低剂量组（27 和13.5 g生药/kg体重）、桂龙咳喘宁胶囊对照组（0.41 g/kg体重）,每组8只.除正常组外以卵蛋白致敏并吸入激发法制备大鼠支气管哮喘模型,各治疗组均从第1次哮喘激发开始（造模第3周）至处死前每天灌胃给药,激发并给药4 w后处死大鼠,取肺组织HE染色,彩色图像分析仪测量支气管管壁厚度、平滑肌厚度,采用ELISA双抗体夹心法测定肺组织MMP-9、TIMP-1含量.结果 与正常组比较,模型组大鼠支气管管壁和平滑肌厚度、肺组织MMP-9、TIMP-1含量及二者比值明显增加（P＜0.01）;与模型组比较,各治疗组均可显著降低支气管管壁和平滑肌厚度（P＜0.01）,降低肺组织MMP-9、TIMP-1含量以及二者比值（P＜0.05或P＜0.01）;且咳喘宁高、低剂量组优于桂龙咳喘宁胶囊组（P＜0.05或P＜0.01）.结论咳喘宁可通过降低肺组织MMP-9和TIMP-1含量,调节二者比值,抑制支气管哮喘大鼠气道壁增厚和平滑肌增生肥大,从而抑制支气管哮喘大鼠气道重塑.     

罗格列酮对兔动脉粥样硬化MMP-2、TIMP-2表达的影响

目的:通过比较罗格列酮和辛伐他汀对兔动脉粥样硬化基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶抑制因子-2(TIMP-2)表达的影响,探讨罗格列酮抗动脉粥样硬化的作用机制。方法:将36只雄性新西兰兔随机分为对照组、模型组、辛伐他汀组和罗格列酮组;采用皮下注射同型半胱氨酸硫内酯及高脂饲养的方法建立兔动脉粥样硬化模型。检测血清氧化低密度脂蛋白(ox-LDL)、sCD40L水平和主动脉粥样硬化组织中MMP-2和TIMP-2的表达。结果:与对照组相比,其他3组MMP-2蛋白表达显著升高,而TIMP-2蛋白表达显著降低;与模型组比较,辛伐他汀和罗格列酮组MMP-2表达显著降低,TIMP-2表达显著增加,血清ox-LDL、sCD40L水平显著降低。辛伐他汀组和罗格列酮组比较,差异无统计学意义。结论:罗格列酮可通过下调兔主动脉粥样硬化组织中MMP-2、上调TIMP-2的表达,降低血清ox-LDL和sCD40L水平,减轻大动脉内膜的炎性反应,发挥抗动脉粥样硬化的作用。