MMP-2,-9及其抑制物TIMP-1在多囊卵巢综合征患者黄素化颗粒细胞上的表达

目的分析多囊卵巢综合征(PCOS)患者黄素化颗粒细胞上基质金属蛋白酶-2,-9(MMP-2,-9)及其组织抑制物TIMP-1的蛋白及mRNA在不同培养时间的表达水平,了解与细胞外基质合成、降解相关的MMPs/TIMPs系统参与PCOS的病理生理机制。方法收集2011年9月到2012年1月行体外受精(IVF)或卵胞浆内单精子注射(ICSI)治疗的PCOS患者30例,对照组为同期行IVF或ICSI治疗的不孕症患者30例。密度梯度法获得卵巢黄素化颗粒细胞,细胞贴壁培养,分别在培养24、48和72h收集颗粒细胞。采用Western blotting方法检测颗粒细胞中MMP-2、MMP-9和TIMP-1的蛋白表达;RT-PCR检测颗粒细胞中MMP-2、MMP-9和TIMP-1mRNA的表达。结果 (1)PCOS组黄素化颗粒细胞MMP-2,-9蛋白表达在各培养时间点(24、48、72h)均显著高于对照组(P均0.05);且MMP-2蛋白表达水平在48、72h相对于24h的比值,以及MMP-9蛋白表达水平在72h相对于24h的比值,PCOS组均显著高于对照组(P均0.01)。(2)PCOS组MMP-2mRNA在24、48、72h的表达,MMP-9mRNA在48、72h的表达均高于对照组,差异有统计学意义(P均0.05);MMP-2,-9mRNA表达水平在48、72h相对于24h的比值,PCOS组高于对照组,差异均有统计学意义(P均0.05)。(3)PCOS组TIMP-1蛋白及mRNA表达在24、48、72h与对照组比较无统计学差异(P0.05)。结论 PCOS患者黄素化颗粒细胞中MMP-2和MMP-9的表达升高,其MMPs/TIMPs的平衡状态向蛋白酶的活性增强方向移动,可能与PCOS患者黄体功能不足有关。     

多囊卵巢综合征患者血清Omentin、CTRP9、Vaspin表达水平及临床意义

目的 观察多囊卵巢综合征(PCOS)患者血清Omentin、CTRP9、Vaspin的表达水平,研究其临床意义。方法 选取2020年1—12月于邯郸市中心医院生殖医学科诊断为PCOS的62例患者为PCOS组,另选同期在本院进行健康体检的65例健康育龄女性作为对照组,采集所有研究对象的空腹外周静脉血,检测其激素指标、糖代谢指标及脂代谢指标等临床相关指标;通过酶联免疫实验(ELISA)检测血清中Omentin、CTRP9、Vaspin水平,并分析与临床指标的相关性;绘制受试者工作特征曲线(ROC曲线),根据曲线下面积AUC分析Omentin、CTRP9、Vaspin对PCOS的诊断价值。结果 (1)临床相关指标显示,PCOS患者体质量指数(BMI)、血清睾酮(T)、空腹血糖(FPG)、空腹胰岛素(FINS)水平、胰岛素抵抗指数HOMA-IR,以及总胆固醇(TC)、甘油三酯(TG)等指标值均显著高于对照组(P<0.05)。(2)ELISA检测结果表明,与对照组相比,PCOS患者Omentin、CTRP9水平显著下降(P<0.05),Vaspin水平显著升高(P<0.05)。...     

阿托伐他汀对急性冠脉综合征患者血清MMP-9、MMP-2及hs-CRP的影响

目的研究应用阿托伐他汀对急性冠脉综合征患者血清MMP-9、MMP-2及hs-CRP的影响。方法选择ACS患者61例,稳定型心绞痛患者19例及正常对照组30例。随机将ACS患者分为阿托伐他汀组（33例）和常规组（28例）,比较各组间血清MMP-9、MMP-2及hs-CRP的水平变化。结果 ACS组与SA组及正常对照组之间血清MMP-9、MMP-2及hs-CRP水平相比差异有统计学意义,两组干预后血清MMP-9、MMP-2及hs-CRP水平相比差异有统计学意义（P〈0.05）。结论阿托伐他汀可降低ACS患者血清MMP-9、MMP-2及hs-CRP水平,减少冠状动脉粥样斑块基质成分的降解和炎症反应,从而起到稳定动脉粥样硬化斑块的作用。     

中西医结合治疗多囊卵巢综合征体会

目的探讨补肾健脾化痰活血方联合克罗米酚治疗多囊卵巢综合征的效果。方法将75例多囊卵巢综合征患者随机分为2组,观察组38例接受补肾健脾化痰活血方联合克罗米酚治疗,对照组37例患者接受克罗米酚治疗。结果观察组妊娠率高于对照组,治疗后LH、T、E_2较对照组降低,FSH较对照组增加,差异均有统计学意义(P0.05)。结论补肾健脾化痰活血方联合克罗米酚治疗多囊卵巢综合征效果肯定,能有效提高患者妊娠率。     

血清IL-9、IL-18及IL-32水平对多发性骨髓瘤患者肾损伤的诊断价值

目的 探讨多发性骨髓瘤(multiple myeloma,MM)患者血清白细胞介素-9(inter-leukin-9,IL-9)、白细胞介素-18(interleukin-18,IL-18)及白细胞介素-32(interleukin-32,IL-32)的表达及其预测肾损伤的价值.方法 选取2016年1月至2019年12...     

慢性肾衰竭患者血清MMP-9和TIMP-1表达及意义

目的检测慢性肾衰竭（CRF）患者血清基质金属蛋白酶-9（MMP-9）及金属蛋白酶组织抑制因子-1（TIMP-1）表达状况,拟在探讨其与肾脏纤维化的关系。方法采用ELISA法测定60例CRF患者（氮质血症期15例,肾衰竭期24例,尿毒症终末期21例）和20例正常对照组血清MMP-9、TIMP-1水平,同时收集患者临床资料,分析血清MMP-9、TIMP-1水平和MMP-9／TIMP-1比值与肾功能损害等临床指标之间的相关性。结果CRF患者各期血清MMP-9、TIMP-1水平明显升高,MMP-9／TIMP-1比值显著降低,随着肾小球滤过率（GFR）下降,MMP-9、TIMP-1呈降低趋势,而MMP-9／TIMP-1比值却逐渐上升。血清TIMP-1水平和MMP-9／TIMP-1比值与血肌酐、GFR及血红蛋白呈高度相关性。结论血清MMP-9、TIMP-1参与了CRF的进展,血清TIMP-1水平或MMP-9／TIMP-1比值能较好地反映肾脏纤维化的程度。     

HOXA10基因在多囊卵巢综合征患者子宫内膜的表达

目的了解同源框基因A10（HOXA10）和胰岛素（INS）在多囊卵巢综合征（PCOS）患者子宫内膜的表达,探讨PCOS患者子宫内膜功能异常的机制。方法采集PCOS患者50例（PCOS组）子宫内膜,分为高胰岛素组（bINS,25例）和高睾酮组（hT,25例）;对照组20例取自月经周期规律妇女的增殖期子宫内膜。采用免疫组化、Western-blot和逆转录聚合酶链反应（RT—PCR）方法检测HOXA10、INS蛋白及mRNA在两组子宫内膜的表达。结果（1）免疫组化结果显示,HOXA00在PCOS组的表达较对照组明显减弱（P〈0．01）;而INS表达却均明显增高（P〈0．01）。（2）Western-blot结果也显示,HOXA10在PCOS组（包括hINS组和hT组）的表达显著低于对照组（P均〈0．01）,而INS的表达显著高于对照组（P均〈0．01）。hINS组与hT组比较,HOXA10表达更显著降低（P〈0．05）;而INS的表达水平在两组之间无显著差异（P〉0．05）。（3）RT-PCR结果显示,HOXA10mRNA的表达在PCOS组（包括hINS组和hT组）显著低于对照组（P均〈0．01）,而INSmRNA表达显著高于对照组（P均〈0．05）。结论PCOS患者的子宫内膜呈现HOXA10表达降低和INS表达增高的状态,可能是该类患者子宫内膜功能异常的原因之一。     

慢性前列腺炎患者前列腺液中IL-2、IL-8及IL-10水平分析

目的: 检测慢性前列腺炎(CP)患者前列腺液中细胞因子IL 2、IL 8及IL 10的水平,探讨这些细胞因子对CP发病机制及诊断方面的价值。　方法: 采用双抗体夹心ELISA法测定 31例CP患者前列腺液中IL 2、IL 8及IL 10的水平,并以 10例健康男性为对照。对每例患者进行两杯法尿液细菌培养、前列腺常规检查和美国国立卫生院前列腺炎症状指数评分 (NIH CPSI)。按NIH分类法将 31例CP患者分为 3型:Ⅱ型 5例,ⅢA型 13例,ⅢB型 13例。　结果: CP组与对照组比较,前列腺液IL 8含量显著升高(P<0. 05),IL 2和IL 10含量显著降低 (P均<0. 05)。Ⅱ型CP组前列腺液IL 2、IL 8和IL 10水平与ⅢA型CP组比较,差异均无显著性。ⅢB型CP组与Ⅱ型和ⅢA型CP组比较,前列腺液IL 8水平显著降低(P均<0. 05)。CP患者前列腺液IL 8水平与IL 10水平呈负相关(r=-0. 503,P<0. 05),与前列腺液白细胞计数呈正相关(r=0. 663,P<0. 05)。前列腺液IL 2、IL 8和IL 10水平与NIH CPSI评分无相关性(P>0. 05)。　结论: 前列腺液中IL 2、IL 8及IL 10在CP的发病过程中起重要作用,是诊断CP有价值的指标。     

胃癌组织中E-钙黏蛋白、MMP-2蛋白和MMP-9蛋白的表达及意义

目的探索胃癌组织及癌旁组织中E-钙黏蛋白、基质金属蛋白酶-2（MMP-2）及基质金属蛋白酶-9（MMP-9）的表达情况,并分析其表达与胃癌临床病理特征的关系。方法采用ABC免疫组化染色方法检测40例胃癌组织和癌旁组织中E-钙黏蛋白、MMP-2蛋白及MMP-9蛋白的表达情况,比较2种组织中3种蛋白的表达情况;并分析该3种蛋白与胃癌临床病理特征的关系。结果胃癌组织与癌旁组织比较,其E-钙黏蛋白的表达下调（P〈0．05）,而MMP．2蛋白和MMP．9蛋白表达均上调（P〈0．05）。T3＋T4期、N1～N3期及Ⅲ＋Ⅳ期胃癌组织中E-钙黏蛋白的表达阳性率分别低于T1＋T2期、NO期及Ⅰ＋Ⅱ期胃癌组织（P〈0．05）;MI期和Ⅲ＋Ⅳ期胃癌组织中MMP-2蛋白的表达阳性率分别高于M0期和I＋Ⅱ期胃癌组织（P〈0．05）;T3＋T4期、Ⅲ＋Ⅳ期及低-未分化的胃癌组织其MMP．9蛋白的表达阳性率分别高于T1＋T2期、I＋Ⅱ期及高-中分化胃癌组织（P〈O．05）。结论E-钙黏蛋白可能参与胃癌的浸润及淋巴结转移,MMP-2蛋白可能参与胃癌的远处转移,而MMP-9蛋白可能参与胃癌的分化及肿瘤浸润。该3种蛋白可作为判断胃癌分期和预后的指标。     

膀胱癌血清中MMP-9与VEGF水平检测及意义

目的　检测基质金属蛋白酶 9(MMP 9)与血管内皮生长因子 (VEGF)在膀胱癌患者血清中的水平 ,探讨其与肿瘤的分级、分期的关系。　方法　采用sandwich ELISA法检测 5 8例膀胱癌患者血清中MMP 9和VEGF水平 ,正常对照组 4 5例。　结果　膀胱癌患者血清中MMP 9和VEGF的水平分别为 737 12 μg/L和 114 8 88ng/L ,均显著高于对照组 4 2 3 5 1μg/L和 84 6 96ng/L(P <0 0 1)。两者的水平与肿瘤的分级、分期有关 ,其中局部浸润性癌显著高于浅表性癌 (P <0 0 1) ;远处转移组显著高于局部浸润组 (P <0 0 1) ;而浅表性癌与对照组无差别 (P >0 0 5 )。G3 级患者MMP 9和VEGF水平均显著高于G1和G2 级 (P <0 0 1)。　结论　膀胱癌患者血清中MMP 9和VEGF水平显著升高 ,并与肿瘤的分级、分期及远处转移有关 ,提示检测两者血清水平有助于对膀胱癌恶性程度的判断。     

克罗米芬/人绝经期促性腺激素小剂量刺激法在多囊卵巢综合征患者促排卵的应用

探讨克罗米芬 (CC) /人绝经期促性腺激素 (HMG)单次用药联合方案 (简称小剂量刺激法 )在多囊卵巢综合征患者中的疗效。对 38例共 45个周期采用 CC5 0 mg/ d×5 d后单次给予 HMG1 5 0 IU的联合方案的卵泡发育情况、排卵及妊娠情况进行了监测 ,并与 CC 32例 43个周期相比较。结果小剂量组妊娠率 34% (1 3/ 38) ,周期排卵率 84%(38/ 45 ) ,卵巢过度刺激综合征 (OHSS)发生 1例 ;CC组妊娠率 1 2 % (3/ 2 4) ,周期排卵率5 6 % (2 4/ 43) ,OHSS1例。结果表明小剂量刺激法是一种安全有效的诱导排卵方案。     

来曲唑或氯米芬联合人绝经期促性腺激素用于PCOS妇女促排卵疗效的比较

目的评估来曲唑(LE)或氯米芬(CC)联合人绝经期促性腺激素(HMG)用于多囊卵巢综合征(PCOS)妇女促排卵的效果。方法回顾性分析接受促排卵治疗的211例PCOS不孕妇女,按促排卵方案分为LE+HMG、CC+HMG以及单用HMG组,比较三组排卵率、妊娠率、单卵泡发育率、促排卵时间以及HMG剂量的差异。结果 LE+HMG组排卵率为91.5%,CC+HMG组86.8%,HMG组71.7%,LE+HMG和CC+HMG组显著高于单用HMG组(P<0.01);LE+HMG组单卵泡发育率显著高于HMG组(85.3%vs.60.5%,P<0.01),但CC+HMG与其他两组比较无统计学差异(P>0.05);LE+HMG、CC+HMG、HMG三组患者的妊娠率分别为25.3%、22.7%和26.3%,组间无统计学差异(P>0.05);LE+HMG组促排卵时间为(15.07±2.41)d,CC+HMG组为(15.27±2.47)d,HMG组为(16.91±2.66)d,三组中HMG剂量分别为(482.3±168.2)U、(486±169.0)U和(595.5±196.6)U,LE+HMG和CC+HMG组促排卵时间及HMG剂量均显著低于HMG组(P<0.01)。结论 LE或CC联合HMG不仅缩短促排卵时间和减少HMG用量,并且提高了排卵率;此外,LE联合HMG更易得到单优势卵泡发育。     

Differential release of matrix metalloproteinase-9 and nitric oxide following infusion of endotoxin to human volunteers

BACKGROUND: Roughly 400.000 cases of sepsis occur every year in the United States only and this is associated with a very high mortality. Bacterial lipopolysaccharide (LPS) triggers systemic inflammatory reactions in sepsis. However, down-stream cellular cascade initiated by LPS is still being elucidated. Nitric oxide (NO) and matrix metalloproteinases-2 and -9 (MMP-2 and MMP-9) are known to be induced by LPS. We have investigated the release of NO, MMP-2 and MMP-9 following infusion of LPS to volunteers. METHODS: IPS (2 ng kg-1) was infused to 10 healthy volunteers. Before the experiments were started the subjects had an intravenous catheters placed. An electrocardiogram was also placed and monitored constantly. Body temperature was measured by ear thermometer every 10 min Venous blood was collected and cell-free plasma assayed for the presence of MMP-2 and MMP-9 using zymography and NO using HPLC assay for NO metabolites, nitrite and nitrate. RESULTS: The administration of LPS resulted in increased body temperature and tachycardia. Time-dependent release of MMP-9(30 fold increase from the baseline) peaking at 2 h following infusion of LPS was observed. LPS did not significantly modify the activity of MMP-2 (P > 0.05). Infusion of LPS did not significantly change the levels of nitrite and nitrate (from 60 +/- 11 to 67 +/- 10 micro m, P > 0.05). CONCLUSION: The release of MMP-9, but not MMP-2 or NO, is a sensitive index of endotoxaemia in humans. MMP-9 release may contribute to the pathogenesis of sepsis via its pro-inflammatory effects on the vasculature.     

Increased levels of interleukin-6 and matrix metalloproteinase-9 are of cardiac origin in acute coronary syndrome

Objective--After off-pump coronary artery bypass (OPCAB) haemostasis might be better preserved compared with on-pump coronary artery bypass grafting (CABG). The aim of this study was to investigate whether this possibly better preserved haemostasis results in a procoagulant activity of the platelets. Design--Thirty patients were studied prospectively, 15 undergoing on-pump CABG and 15 undergoing OPCAB. Platelet function was evaluated four times within the first 24?h: preoperatively, postoperatively, 4?h and 1 day after surgery with a bedside whole blood clotting test. Results--A significant increase of platelet-activating-factor-induced platelet aggregation was observed postoperatively after OPCAB (p?&lt;?0.01). Only two patients did not reach preoperative values within 1 day postoperatively and four patients had a more than twofold increase. Platelet aggregation immediately after on-pump CABG was reduced to near half of preoperative values, but within 1 day postoperatively normal platelet aggregation was regained in half of the patients. Conclusion--This study has mainly indicated that platelets after OPCAB were more easily activated in the early postoperative period. After CABG with cardiopulmonary bypass we found a temporary platelet dysfunction which seemed to be overcome within the first postoperative day.     

Inhibition of inducible nitric oxide synthase and superoxide production reduces matrix metalloproteinase-9 activity and restores coronary vasomotor function in rat cardiac allografts.

OBJECTIVE: Oxidants such as nitric oxide (NO) and superoxide are involved in coronary endothelial dysfunction, an early event in the process of allograft coronary atherogenesis, possibly by activation of matrix metalloproteinases (MMPs) and extracellular matrix proteins. We investigated the contribution of inducible nitric oxide synthase (iNOS) derived NO and superoxide on (MMP)-9 activity and to changes in coronary vasomotor function in rat cardiac allografts. METHODS AND RESULTS: An allogenic (Brown Norway to Lewis rats) heterotopic cardiac transplantation model was used to study the effect of continuous treatment with a selective iNOS inhibitor; N-(3-(aminomethyl) benzyl) acetamidine (1400W), and polyethylene glycol conjugated superoxide dismutase (SOD) either alone or in combination on coronary vasomotor dysfunction. 1400W or SOD 24 h alone or their combination improved endothelium-dependent (bradykinin) and -independent (sodium nitroprusside) coronary flow reserve and inhibited enhanced MMP-9 protein and activity. In addition, histopathological study revealed that either 1400W or SOD or their combination reduced superoxide production and nitrotyrosine protein. CONCLUSION: The present study demonstrates for the first time that selective iNOS inhibition or SOD treatment reduces enhanced MMP-9 protein and activity associated with improvement of both, endothelium-dependent and -independent coronary vasomotor function in rat cardiac allografts. This is accompanied by reduction of nitrotyrosine and superoxide production. This suggests that the proteolytic enzyme MMP-9 is an effector molecule of oxidant-mediated coronary vasomotor dysfunction.     

一氧化氮增加人软骨肉瘤细胞表达基质金属蛋白酶-13的机制研究

目的研究一氧化氮(NO)是否通过MAPK和NF-κB通路增加基质金属蛋白酶-13(MMP-13)的表达,探讨骨关节炎(OA)发病机制中NO的作用。方法购买并传代人软骨肉瘤细胞(SW1353),用不同浓度的NO供体MAHMA-NONOate刺激后检测MMP-13以及MAPK和NF-κB通路中的蛋白激酶的表达水平。用不同浓度的MAPK和NF-κB通路中的蛋白激酶的抑制剂预先处理细胞后,再用MAHMA-NONOate刺激细胞,再次检测MMP-13以及MAPK和NF-κB通路中的蛋白激酶的表达水平。结果 MAHMA-NONOate增加了细胞MMP-13的表达水平,同时也增加了MAPK和NF-κB通路中的蛋白激酶的活性水平,MAPK家族中的JNK的抑制剂SP600125可以抑制MMP-13的表达水平,NF-κB的抑制剂SN5O也可以抑制MMP-13的表达水平。结论 NO可以通过MAPK家族中的JNK和NF-κB通路增加MMP-13的表达。     

Correlation between interleukin-6 and matrix metalloproteinase-9 in early wound healing in children

Interleukin-6 and matrix metalloproteinase-9 concentrations in the wound fluid and their associations to cellular changes were determined in early wound healing. Wound healing of 75 children who underwent elective operations was studied with the Cellstick(R) device, which was inserted into the wound at the end of the operation and removed 3 or 24 hours post-wounding. Differential counts of the wound cells and interleukin-6 and matrix metalloproteinase-9 concentrations in the wound fluid were analyzed. Interleukin-6 and the matrix metalloproteinase-9 concentrations increased in parallel (r = 0.81). The proportion of wound neutrophils increased (p < 0.0001) and lymphocytes decreased (p < 0. 0001) between the observation times. The number of wound neutrophils had a strong correlation with both interleukin-6 (adjusted R2 = 0.41, p < 0.0001) and matrix metalloproteinase-9 concentrations (adjusted R2 = 0.37, p < 0.0001). The extracellular matrix degradation process of the early wound healing seems to be closely linked to the inflammatory response. Both of these measured markers are associated significantly with the neutrophil proportion in the wound.     

Nitric oxide modulates expression of matrix metalloproteinase-9 in rat mesangial cells

Nitric oxide modulates expression of matrix metalloproteinase-9 in rat mesangial cells. BACKGROUND: High-output levels of nitric oxide (NO) are produced by rat mesangial cells (MCs) in response to proinflammatory cytokines such as interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) by the inducible isoform of NO synthase (iNOS). We tested modulatory effects of NO on the expression and activities of matrix metalloproteinases-9 and -2 (MMP-9 and MMP-2), respectively. Temporal and spatial expression of these MMPs and their specific inhibitors, the tissue inhibitors of metalloproteinases (TIMPs), seems to be critical in the extensive extracellular matrix (ECM) remodeling that accompanies sclerotic processes of the mesangium. Methods and Results. Using the NO donors S-Nitroso-N-acetyl-D,L-penicillamine (SNAP) and DETA-NONOate, we found strong inhibitory effects of NO mainly on the IL-1beta-induced MMP-9 mRNA levels. NO on its own had only weak effects on the expression of MMP-9 and MMP-2. The addition of the NOS inhibitor NG-monomethyl L-arginine (L-NMMA) dose dependently increased steady-state mRNA levels of cytokine-induced MMP-9, suggesting that endogenously produced NO exerts tonic inhibition of MMP-9 expression. MMP-9 activity in conditioned media from MCs costimulated with IL-1beta and NO donor contained less gelatinolytic activity than media of cells treated with IL-1beta alone. Exogenously added NO did not alter gelatinolytic activity of MMP-9 in cell-free zymographs. The expression levels of TIMP-1 were affected by NO similarly to the expression of MMP-9. CONCLUSION: We conclude that NO modulates cytokine-mediated expression of MMP-9 and TIMP-1 in rat MCs in culture. Our results provide evidence that NO-mediated attenuation of MMP-9 gelatinolytic activity is primarily due to a reduced expression of MMP-9 mRNA, and not the result of direct inhibition of enzymatic activity.