PRL-3和MMP-9与子宫内膜异位症侵袭、种植关系的研究

目的:观察促肝细胞再生磷酸酶3(PRL-3)和基质金属蛋白酶9(MMP-9)在子宫内膜异位症(EMs)的表达,探讨它与子宫内膜异位症侵袭、种植的关系。方法:用SABC免疫组织化学法检测PRL-3和MMP-9在31例EMs的异位内膜及在位内膜组织、27例正常子宫内膜组织的表达。结果:PRL-3与MMP-9表达的强弱顺序均为:EMs组异位内膜>EMs组在位内膜>对照组子宫内膜,组间差异有统计学意义(P<0.05);Ⅰ~Ⅱ期的异位内膜PRL-3及MMP-9表达强度均高于Ⅲ~Ⅳ期(P<0.05),但在位内膜二者表达强度在不同临床分期中无统计学差异(P>0.05)。EMs组异位内膜的PRL-3与MMP-9表达呈正相关(P<0.05),在位内膜的PRL-3与MMP-9表达则无明显相关性(P>0.05)。结论:PRL-3和MMP-9可能与EMs的侵袭、种植密切相关。     

压力性尿失禁患者阴道壁结缔组织MMP-2及其抑制物的表达

目的:探讨女性压力性尿失禁(SUI)患者阴道壁结缔组织中基质金属蛋白酶-2(MMP-2)和基质金属蛋白酶组织抑制物-2(TIMP-2)mRNA表达与SUI发病的关系。方法:7例无SUI妇女为对照组(A组),24例有完整随访资料的女性SUI患者的阴道壁结缔组织标本为研究组,分为<60岁组(B组)和>60岁组(C组)。应用逆转录-聚合酶链反应分别检测MMP-2和TIMP-2表达量。结果:SUI组患者的MMP-2均较非SUI组显著升高,差别有显著性(P<0.01),TIMP-2均显著下降,差异有统计学意义(P<0.01)。MMP-2在B组和C组之间的差异无显著性(P>0.05),TIMP-2的mRNA表达量C组较B组显著下降,差别有统计学意义(P<0.01)。MMP-2/TIMP-2从A组到B组再到C组显著升高,A组相似文献   

Matrix metalloproteinase-2, membranous type 1 matrix metalloproteinase,and tissue inhibitor of metalloproteinase-2 expression in ectopic and eutopic endometrium

OBJECTIVE: To investigate expression of matrix metalloproteinase-2 (MMP-2), membranous type 1 matrix metalloproteinase (MT1-MMP), and tissue inhibitor of metalloproteinase-2 (TIMP-2) in ectopic and eutopic endometrium from women with and without endometriosis throughout the menstrual cycle. DESIGN: Molecular studies in human tissue. SETTING: Reproductive immunology laboratory of a university medical center. PATIENT(S): Fifty-three premenopausal woman (23 with endometriosis and 30 without endometriosis) undergoing laparoscopic surgery. Endometrium and ectopic endometriosis tissue were obtained at the time of surgery. MAIN OUTCOME MEASURE(S): Messenger RNA and protein expression from eutopic and ectopic endometrium was analyzed by using quantitative competitive polymerase chain reaction, zymography, and Western blot assay. RESULT(S): Uterine endometrium from women with endometriosis expressed higher levels of MMP-2 and MT1-MMP and lower levels of TIMP-2 than did endometrium from normal women. CONCLUSION(S): Eutopic endometrium from patients with endometriosis may be more invasive and prone to peritoneal implantation because of greater expression of MMP-2 and MT1-MMP and lower expression of TIMP-2 messenger RNA, compared with endometrium from women without endometriosis. Thus, increased proteolytic activity may help to explain the invasive factors that result in endometriosis.     

Hypoxia inhibits invasion of extravillous trophoblast cells through reduction of matrix metalloproteinase (MMP)-2 activation in the early first trimester of human pregnancy

During early pregnancy, extravillous trophoblast (EVT) cells are exposed to very low pO₂ values. In this study, we investigated the proteolytic functions and invasiveness of human primary EVT cells under hypoxic conditions to show the early placental pathophysiology.Placental samples (from 5 to 10 weeks gestation) were obtained at termination of pregnancy. Cytotrophoblast cells were separated by Percoll^® gradient method and cultured on Matrigel^® to obtain an invasive phenotype (similar to EVT). The invasion capacity (Matrigel-coated invasion assay), migration of the cells (wound healing assay), activity and expression of matrix metalloproteinase (MMP)-2 and tissue inhibitor for MMP (TIMP)-2 (gelatin gel zymography, ELISA, and quantitative RT-PCR), and expression of membrane-type (MT)1-MMP (western blot) were investigated. All cultures (except for quantitative RT-PCR) were performed under 20% oxygen, 5% oxygen, and 5% oxygen with 3 repetitions of 0.1% oxygen hypoxic stimulation for 1 h.Invasion and MMP2 activity of the cells were significantly increased in 20% and decreased in 0.1% oxygen. There was no significant difference in cell migration among the oxygen environments. Concentrations of MMP2 in the supernatant and expression of MT1-MMP were increased in both the 0.1% and 20% oxygen environments. The MMP2 mRNA level was increased after 1-h stimulation with 0.1% oxygen. The TIMP2 concentration was increased only in 20% oxygen, but the mRNA level was decreased in 0.1% oxygen.These results suggested that hypoxia might inhibit the invasive capacity and MMP2 activation of EVT cells in the early first trimester of pregnancy. Decrease in TIMP2 production may reduce the MMP2/TIMP2/MT1-MMP complex and lead to this unique behavior of EVT cells under hypoxic conditions.     

呼吸道异物患儿肺泡灌洗液中羟脯氨酸基质金属蛋白酶9及其抑制物水平的研究

目的以肺泡灌洗液(BALF)中羟脯氨酸(HYP)、基质金属蛋白酶9(MMP-9)及其抑制物(TIMP-1)为观察指标，了解呼吸道异物吸入对气道损害、重建的影响。 方法对2002年6月至2003年3月在浙江大学儿童医院住院的46例呼吸道异物患儿，按异物滞留体内时间分为3组(A组：<7d，B组：~30d，C组：>30d)，对3组患儿的BALF进行细胞学分析，并对其中的HYP、MMP-9细胞阳性率和TIMP-1质量浓度进行测定。对照组为12例有异物吸入可疑病史，但最后排除的儿童。 结果对照组与患儿组间细胞学计数差异无显著性(P>0.05)，但肥大细胞仅在患儿BALF中发现。对照组及各组患儿的MMP-9细胞阳性率分别为33.3%、62.5%、80.0%和93.3%，TIMP-1细胞阳性率分别为16.3%、50.0%、80.0%和93.3%，总体差异均有显著性意义(P=0.006和P<0.001)；各组间HYP差异有显著性意义(P<0.001)。 结论气道异物促进MMP-9、TIMP-1及HYP的表达和分泌，提示气道异物可导致支气管肺组织破坏和重建，其程度与异物滞留时间有关。     

持续低血糖新生大鼠大脑基质金属蛋白酶-2活性和氧化型谷胱甘肽水平的变化

目的 研究7日龄新生大鼠持续低血糖对脑组织氧化应激和基质金属蛋白酶-2(matrix metalloproteinase-2,MMP-2)水平的影响,探讨低血糖引起新生儿脑损伤的机制. 方法 7日龄大鼠皮下注射普通胰岛素诱导持续性低血糖(n=6),观察36 h后收集脑皮质、海马和丘脑组织,采用谷胱甘肽活性测定试剂盒测定总谷胱甘肽( glutathione,GSH)和氧化型GSH( glutathione disulfide,GSSG)含量,计算GSSG/GSH,并采用酶谱法测定MMP-2活性.设正常血糖对照组6只.统计学方法采用t检验、双因素方差分析、单因素方差分析或Pearson相关性分析. 结果 持续低血糖36 h后,低血糖组脑组织总GSSG含量较对照组增高约1.5倍[(15.89±5.46)mg/g蛋白与(6.15±3.42) mg/g蛋白,t=3.704,P=0.004].GSSG/GSH比率也显著高于对照组[(5.58±1.79)％与(2.79±1.76)％,t=2.712,P=0.022)].低血糖组与对照组比较,大脑皮质和海马部位的GSSG/GSH比率差异无统计学意义,而丘脑GSSG含量和GSSG/GSH比率显著高于对照组[GSSG:(15.93±5.75)mg/g蛋白与(5.03±5.14)mg/g蛋白,P＜0.05; GSSG/GSH:(6.50±3.25)％与(2.41±3.12)％,P＜0.05].低血糖组脑组织MMP-2总活性较对照组显著增高(2.22±0.59与1.21±0.17,t=4.064,P=0.002).低血糖组与对照组脑组织不同部位比较,MMP-2活性均显著增高(皮质:2.14±0.51与1.17±0.27;海马:2.31±0.72与1.22±0.37;丘脑:2.22±0.68与1.24±0.18;三者P均＜0.0i).脑组织MMP-2活性与GSSG的含量和GSSG/GSH比率分别呈正相关(r=0.575和0.484,P=0.0002和0.0003). 结论 氧化应激是7日龄新生大鼠持续低血糖造成脑损伤的重要机制,其相关部位主要为丘脑.低血糖可使脑组织MMP-2活性升高,与GSSG和GSSG/GSH的含量有关.     

Matrix metalloproteinase-9 polymorphism and risk of pelvic organ prolapse in Taiwanese women

Objective

Matrix metalloproteinase-9 is known to play an important role in the pathophysiology of pelvic organ prolapse. We investigated whether the matrix metalloproteinase-9 gene polymorphisms were associated with pelvic organ prolapse by conducting a case–control association study in 92 women with pelvic organ prolapse and 152 women without pelvic organ prolapse.

Study design

Genotypes of the matrix metalloproteinase-9 gene polymorphisms (rs3918242, rs17576, and rs2250889) were determined by polymerase chain reaction, followed by restriction fragment length polymorphism analysis.

Results

There was significant difference between women with and without pelvic organ prolapse in the distribution of the matrix metalloproteinase-9 rs17576 genotypes evaluated. Using multivariable logistic regression, menopausal status, matrix metalloproteinase-9 rs17576 genotype AG, and matrix metalloproteinase-9 rs17576 genotype GG were significantly associated with pelvic organ prolapse.

Conclusion

The present study shows that the polymorphism of matrix metalloproteinase-9 rs17576 may be associated with pelvic organ prolapse.     

MMP-2/TIMP-2在人及裸鼠异位子宫内膜组织中的表达及与雌、孕激素的关系

目的:探讨MMP-2/TIMP-2在子宫内膜异位症发病机制中的作用及雌、孕激素对二者表达的影响。方法:(1)用RT-PCR及SP免疫组化法检测51例异位内膜组织及28例正常子宫内膜组织中MMP-2/TIMP-2mRNA及蛋白的表达;(2)建立裸鼠子宫内膜异位症模型并予以雌、孕激素干扰,随机分成4组(E组、P组、E+P组及对照组C),观察不同激素干扰下裸鼠内异症模型的种植成功率及其大体、镜下的形态表现;用免疫组化法检测MMP-2/TIMP-2蛋白在不同组别裸鼠异位内膜中的表达。结果:(1)四组裸鼠内异症模型中种植物均成活,总种植成功率86.4%(51/59),各组间成功率无显著差异。其中P组种植物数量较其他组明显增多,P组、E+P组种植物明显大于E组、C组。受外源性雌、孕激素影响,镜下种植物分别呈现增生期和分泌期特点;(2)MMP-2/TIMP-2mRNA及蛋白在人及各组裸鼠异位内膜组织中均有表达,MMP-2相对表达量明显高于正常子宫内膜,差异有统计学意义(P<0.05);TIMP-2表达与正常内膜组织则无显著差异,MMP-2/TIMP-2比值明显增高(P均<0.01)。(3)E组、E+P组MMP-2蛋白表达均明显高于C组,而P组与C组无差异;TIMP-2蛋白在C组的表达明显高于其他激素干扰组。结论:裸鼠模型是EMs早期临床研究的理想模型。MMP-2/TIMP-2在人及裸鼠异位组织中异常表达,动态平衡紊乱,在EMs发生发展中起重要作用。雌激素可上调MMP-2蛋白表达,促进异位内膜种植;孕激素未表现对抗雌激素的作用,且通过抑制TIMP-2表达,同样造成MMP-2/TIMP-2比值升高,使异位内膜侵袭性增高,表现出"孕激素抵抗"现象。     

MMP-2和MMP-9参与多种因素对胎盘滋养细胞侵袭力的调控

成功妊娠有赖于滋养细胞对子宫内膜的适度侵袭。侵袭功能出现异常,可导致滋养细胞对子宫内膜侵入过深或过浅、子宫螺旋动脉重塑障碍、胎盘缺氧缺血及母体全身血管内皮细胞损伤,导致多种不良妊娠结局以及妊娠相关疾病的发生。明胶酶基质金属蛋白酶2(matrix metallopeptidase 2,MMP-2)和MMP-9因可降解Ⅳ型胶原参与对滋养细胞侵入子宫内膜能力的调控。研究表明,激素、细胞因子、转录因子、环境污染物以及其他多种物理刺激等均可以影响滋养细胞的侵袭力,并且在探讨其影响机制时都最终以MMP-2和MMP-9表达水平的变化来解释。对影响滋养细胞侵袭力的多种因素进行综述,有助于以MMP-2和MMP-9为靶点的不良妊娠以及妊娠相关疾病的预防或治疗。     

基质金属蛋白酶9及其组织抑制物1的水平变化与胎膜早破发病的关系

目的探讨母血、羊水、脐血中基质金属蛋白酶9(MMP-9)及金属蛋白酶组织抑制物1(TIMP-1)的水平变化与胎膜早破发病的关系。方法采用双抗体夹心酶联免疫吸附法,动态检测58例胎膜早破患者(胎膜早破组)及38例正常孕妇(对照组)的母血、羊水、脐血中MMP-9及TIMP-1的水平,同时进行胎膜组织的病理检查,对合并绒毛膜羊膜炎的患者也进行相应的MMP-9及TIMP-1的水平检测。结果(1)胎膜早破组母血、脐血及羊水中MMP-9水平分别为(141·9±84·6)、(138·2±81·4)及(85·6±27·5)ng/L,其水平均明显高于对照组,两组比较,差异有统计学意义(P<0·05、P<0·05及P<0·01);胎膜早破组母血、羊水、脐血中TIMP-1水平分别为(378·1±220·2)、(44·6±24·0)及(257·2±98·8)ng/L,其水平均明显低于对照组,两组比较,差异有统计学意义(P<0·05、P<0·05及P<0·01)。(2)母血、羊水、脐血中MMP-9的水平随破膜时间的延长而升高,尤其破膜时间超过24h时升高更为明显;而TIMP-1则随破膜时间的延长而下降,尤其破膜时间超过24h时降低更为明显。(3)两组共合并绒毛膜羊膜炎患者15例,其中胎膜早破组13例(13/58,22%),绒毛膜羊膜炎患者的母血、脐血、羊水中MMP-9水平分别为(183·8±84·7)、(171·2±92·9)及(95·5±21·1)ng/L,其水平均明显高于非绒毛膜羊膜炎患者(81例),两者比较,差异有统计学意义(P<0·05、P<0·05及P<0·01)。绒毛膜羊膜炎患者的母血、羊水、脐血中TIMP-1水平分别为(269·7±144·4)、(32·1±16·6)及(210·6±81·9)ng/L,其水平均明显低于非绒毛膜羊膜炎患者,两者比较,差异有统计学意义(P<0·05、P<0·05及P<0·01)。(4)胎膜早破组的新生儿Apgar评分≤7分者,其母血、脐血、羊水中MMP-9水平分别为(234·4±79·4)、(222·1±120·1)及(108·5±42·2)ng/L,其水平均明显高于Apgar评分≥8分者,两者比较,差异有统计学意义(P<0·05、P<0·05及P<0·01);与之相反,胎膜早破组的新生儿Apgar评分≤7分者,母血、羊水、脐血中TIMP-1的水平分别为(225·3±121·7)、(25·2±15·8)及(181·7±135·2)ng/L,其水平均明显低于Apgar评分≥8分者,两者比较,差异有统计学意义(P<0·05、P<0·05及P<0·01)。结论MMP-9水平的异常升高及TIMP-1水平的显著下降,是胎膜早破发病的重要机制。MMP-9及TIMP-1的水平检测可作为一种新的生物学标志物用于胎膜早破并绒毛膜羊膜炎感染的诊断,尤其适用于亚临床感染状态的早期诊断,并有助于评估新生儿预后。     

Increased matrix metalloproteinases-1,-9 in the uterosacral ligaments and vaginal tissue from women with pelvic organ prolapse

Objectives

To investigate the possible association of increased matrix metalloproteinases (MMPs)-1,-9 with pelvic organ prolapse (POP) and to evaluate whether inflammatory processes contribute to its development.

Study design

Forty women who underwent hysterectomy, 20 with POP grade 2 and above, and 20 without POP, participated in the study. Biopsies from the uterosacral ligaments and vaginal mucosa were obtained from each woman. Each biopsy was sectioned and stained for MMP-1 and MMP-9 by immunohistochemical methods and with hematoxylin and eosin (H&E). MMP-1,-9 expressions were evaluated on the immunostained slides. H&E stained sections were examined for possible inflammatory changes.

Results

A higher stromal (extra-cellular) expression of MMPs-1,-9 was found in POP cases compared with controls in vaginal biopsies (MMP-1: p = 0.004; MMP-9: p = 0.042) as well as in uterosacral ligament biopsies (MMP-1: p = 0.011; MMP-9: p = 0.015). Increased intracellular expression of both MMPs was also demonstrated in fibroblasts in biopsies of women with POP (p < 0.001 for all). Most of these differences persisted after controlling for age. The degree of inflammatory changes reflected by the number of lymphocytes, plasma cells and capillary-sized blood vessels per 10 high power fields, was similar in specimens obtained from women with and without POP.

Conclusions

The expression of MMPs-1,-9 appears to be increased in tissues from women with POP. This supports an association, although not a causal relation, between increased MMPs-1,-9 and POP. Inflammation does not seem to play an important role in the pathogenesis of POP.     

早期自然流产患者绒毛KiSS-1、MMP-9和MMP-2基因及其蛋白表达

目的研究早期自然流产患者绒毛肿瘤转移抑制基因KiSS-1与基质金属蛋白酶(MMP-9、MMP-2)mRNA及蛋白的表达,探讨其与自然流产的关系。方法采用半定量RT-PCR、免疫印迹法检测30例正常早期妊娠和30例早期自然流产患者绒毛KiSS-1、MMP-9和MMP-2的mRNA及蛋白水平,用明胶酶谱分析法检测上述样本孵育液MMP-9、MMP-2的酶活性。结果(1)自然流产患者绒毛组织中MMP-9及MMP-2mRNA表达水平低于正常早期妊娠组(P<0.05),KiSS-1mRNA的表达高于正常早期妊娠组差异有统计学意义(P<0.05);(2)Westernblotting分析显示自然流产患者绒毛MMP-9、MMP-2蛋白表达低于正常早期妊娠差异有统计学意义(P<0.05),KiSS-1蛋白表达高于正常早期妊娠差异有统计学意义(P<0.05);(3)明胶酶谱分析结果显示,自然流产患者绒毛MMP-9、MMP-2酶活性低于正常早期妊娠者差异有统计学意义(P<0.05)。结论自然流产患者绒毛促浸润基因MMP-9、MMP-2mRNA及其蛋白表达水平降低,同时抑制浸润基因KiSS-1mRNA及其蛋白表达增加,造成滋养细胞浸润能力下降,可能与自然流产的发生有关。     

同型半胱氨酸对滋养细胞基质金属蛋白酶2、9表达的影响

目的探讨同型半胱氨酸（Hcy）对早孕期滋养细胞基质金属蛋白酶（MMP）2、9表达的影响及其在子病前期发病过程中的作用。方法分离培养早孕期滋养细胞,用Hcy刺激早孕期滋养细胞48h,对照组Hcy刺激浓度为0mmol／L,实验组的刺激浓度为1mmol／L;以实时荧光定量RT-PCR检测早孕期滋养细胞MMP-2、MMP-9mRNA的表达变化;明胶酶谱分析法测定早孕期滋养细胞MMP-2、MMP-9酶活性变化,以积分吸光度值表示。结果实验组与对照组比较,早孕期滋养细胞MMP-2mRNA表达降低21％,MMP-9mRNA表达降低11％;实验组与对照组比较,MMP-2蛋白表达降低14％,MMP-9蛋白表达降低52％。结论Hey能够抑制早孕期滋养细胞MMP02、MMP-9的表达,从而影响滋养细胞侵袭性。     

Raised serum levels of matrix metalloproteinase-9 in women with polycystic ovary syndrome and its association with insulin-like growth factor binding protein-1

Background.?It has been suggested in recent studies that matrix metalloproteinases (MMPs) may be implicated in the pathogenesis of polycystic ovary syndrome (PCOS) through regulating ovarian tissue remodeling. In addition to degrading the extracellular matrix, MMPs exhibit the ability to cleave insulin-like growth factor binding protein-1 (IGFBP-1), the major regulator of insulin-like growth factor-I (IGF-I) in serum. The present study aimed to investigate the possible role of MMPs in the pathophysiology of PCOS.

Methods.?Serum levels of MMP-9, tissue inhibitor of metalloproteinase-1 (TIMP-1), IGF-I and IGFBP-1 were measured in 42 patients with PCOS and 30 healthy women with regular menstruation, matched for age and body mass index. Correlation between IGFBP-1 and other parameters in the PCOS group was analyzed by Pearson's linear correlations.

Results.?Serum MMP-9 concentrations and MMP-9/TIMP-1 ratios were significantly higher in PCOS women than in controls. Serum levels of IGFBP-1 were markedly lower in the PCOS group. There was a negative correlation between serum IGFBP-1 and MMP-9 in women with PCOS.

Conclusion.?Our results raise the possibility that MMPs may be implicated in the pathophysiology of PCOS either by regulating ovarian tissue remodeling or indirectly by facilitating IGF-I bioavailability through proteolysis of IGFBP-1.     

MMP-2、TIMP-2在子宫内膜异位症组织中的表达及其意义

目的研究基质金属蛋白酶-2(MMP-2)及其抑制因子-2(TIMP-2)mRNA在子宫内膜异位症异位和在位内膜组织中的表达及临床意义。方法分别选取子宫内膜异位症异位内膜组织50例、在位内膜组织26例和对照组正常子宫内膜组织22例为研究对象,应用半定量逆转录聚合酶链反应技术(RT-PCR)检测MMP-2、TIMP-2mRNA的表达。结果(1)MMP-2mRNA在异位症异位内膜组织中相对表达量明显高于正常子宫内膜组织和在位内膜,差异均有显著性(P<0.01,P<0.01);TIMP-2mRNA在子宫内膜异位症异位内膜和在位内膜组织中均低表达,其相对表达量分别明显低于正常子宫内膜,差异均有显著性(P<0.01;P<0.01);(2)异位症异位内膜组织中MMP-2mRNA、TIMP-2的表达与r-AFS临床分期无关,差异无显著性(P>0.05)。结论异位和在位内膜组织中MMP-2高表达,TIMP-2低表达,使MMP-2与TIMP-2平衡失调,异位内膜组织侵袭降解ECM的能力增强,从而在子宫内膜异位症中的发生发展中起了重要作用。     

白介素24在子痫前期胎盘的表达及与基质金属蛋白酶2关系的研究

目的:研究IL-24在子痫前期胎盘的表达及rhIL-24对TEV-1细胞MMP-2蛋白活性表达的影响,探讨二者与子痫前期发病机制的关系。方法:RT-PCR检测子痫前期及正常足月孕妇胎盘中IL-24 mRNA的表达;明胶酶谱检测0、10、50、100、500ng/mlrhIL-24作用后TEV-1细胞的MMP-2活性;Western blot检测不同浓度rhIL-24作用后TEV-1细胞的MMP-2蛋白表达。结果:子痫前期组胎盘组织中IL-24 mRNA表达显著高于正常足月组(P<0.05)。当rhIL-24浓度≥50ng/ml后,TEV-1细胞中MMP-2蛋白活性表达均明显降低(P<0.05),并呈一定的浓度依赖性。结论:IL-24异常升高,抑制了滋养细胞MMP-2的分泌及活性,使其侵袭力下降,从而在子痫前期发病中起一定作用。