基质金属蛋白酶-9及组织抑制剂-1与肾小管间质纤维化

肾小管间质纤维化（TIF）是所有慢性肾脏疾病（CKD）进展到终末期肾病（ESRD）的共同途径,其主要病理特征为肾间质成纤维细胞的增生和细胞外基质（如I、Ⅱ、Ⅲ、Ⅳ型胶原、纤维连接蛋白、层粘连蛋白）的过度积聚。基质金属蛋白蓐每／基质金属蛋白酶组织抑制剂（MMPs／TIMPs）是调控细胞外基质（ECM）降解的重要酶系,大量研究表明其在肾小管间质纤维化的发生与发展中起重要作用,其中以MMP-9及TIMP-1最为重要。现对MMP-9及TIMP-1与肾小管间质纤维化关系的研究进展作一综述。     

TIMP-2、MMP-2和MT1-MMP在活化肝星状细胞中的表达及对细胞外基质合成分泌的影响

目的 研究金属蛋白酶组织抑制剂-2(TIMP-2)、基质金属蛋白酶-2(MMP-2)和膜型基质金属蛋白酶-1(MT1-MMP)在活化肝星状细胞(HSCs)中的表达,观察其对细胞外基质(ECM)合成分泌的影响.方法 原代分离培养大鼠HSCs活化后,分别给予40～160 pmol化学合成经修饰抗TIMP-2 siRNA进行干预,检测培养细胞上清液透明质酸(HA)、Ⅲ型前胶原(PCⅢ)和羟脯氨酸(Hyp)的含量,采用荧光实时定量PCR法检测TIMP-2、MMP-2、MT1-MMP、MMP-13、COL Ⅰ和COL Ⅲ mRNA的表达,western印迹检测TIMP-2、MT1-MMP和MMP-13蛋白表达及明胶酶谱法检测MMP-2蛋白表达.结果 应用化学合成经修饰抗TIMP-2 siRNA后,TIMP-2、MMP-2、MT1-MMP、COL Ⅰ和COL Ⅲ的表达明显降低,而MMP-13的表达则明显增加,培养细胞上清液中HA、PCⅢ和Hyp的含量也明显减少.结论 TIMP-2通过MT1-MMP介导MMP-2的活化,抑制TIMP-2的表达,MT1-MMP和MMP-2的表达随之降低,而HSCs合成分泌ECM也相应减少.     

雷帕霉素在大鼠实验性三硝基苯磺酸结肠炎发病中的作用

目的 观察转录激活因子3(STAT3)特异性抑制剂雷帕霉素(RPM)对大鼠实验性三硝基苯磺酸结肠炎的作用,以及对基质金属蛋白酶(MMPs)、基质金属蛋白酶组织抑制剂(TIMPs)表达的影响,探讨信号转导和转录激活因子-3(STAT3)信号转导通路在大鼠结肠炎发病机制中的作用.方法 建实验性大鼠结肠炎模型后,给予RPM腹腔注射治疗一周后处死大鼠,观察结肠炎症改变并做出评分,SYBR Green Ⅰ实时荧光定量RT-PCR检测结肠组织MMP-1、MMP-2、MMP-3、TIMP-1及TIMP-2的mRNA表达.结果 RPM治疗组肠道损伤积分为(5.17±1.80),低于对照组(P<0.05);MMP-1、MMP-2的mRNA表达量在RPM组均明显降低(P<0.05),而MMP-3、TIMP-1和TIMP-2 mRNA表达无明显差异(P>0.05).结论 RPM通过阻断STAT3信号通路的活化能缓解大鼠结肠炎的病情,这一作用可能是通过抑制MMP-1和MMP-2的表达实现的.     

益心方对血管再狭窄小鼠基质金属蛋白酶与组织型基质金属蛋白酶抑制因子表达的影响

目的 探讨再狭窄血管中基质金属蛋白酶(MMPs)与组织型基质金属蛋白酶抑制因子(TIMPs)的表达及益心方对其的影响.方法 制备大鼠血管再狭窄模型,采用不同浓度的益心方药物浓度干预,观察颈总动脉形态学改变,免疫组化法检测颈总动脉MMPs与TIMPs的表达.结果 假手术组血管内膜和中膜处均可见少量散在分布的MMP-2、MMP-9、TIMP-1和TIMP-2的阳性染色表达颗粒,以MMP-2表达最为明显;与假手术组比较,球囊导管内皮剥脱术后,模型组血管中MMP-2、MMP-9、TIMP-1和TIMP-2表达均显著增加(P<0.05或P<0.01),主要在新生内膜中表达,胞浆和基质内出现大量特异性的阳性颗粒;与模型组比较,益心方治疗低、高剂量组能明显抑制血管壁MMP-2、MMP-9的表达(P<0.05或P<0.01),其特异性的阳性颗粒与模型组相比显著减少,对TIMP-1和TIMP-2的表达也有减少的趋势.结论 益心方抑制血管再狭窄,其机制与抑制血管平滑肌细胞MMPs与TIMPs表达有关.     

基质金属蛋白酶及其组织抑制物与肝纤维化

基质金属蛋白酶(matrix metalloproteinases,MMPs)是降解细胞外基质(ECM)的主要蛋白酶。基质金属蛋白酶组织抑制物(tissue inhibitors of metalloproteinases,TIMPs)是组织中MMPs主要的内源性抑制因子,至今共有4种。肝纤维化早期MMPs轻度升高;而肝纤维化的中晚期,MMPs活性则明显降低,TIMPs的表达显著增加。一旦肝纤维化逆转,MMPs的表达又显著增强,而TIMPs的表达逐渐降低。表明MMPs和TIMPs与肝纤维化的发生和发展密切相关。     

氯沙坦对高血压大鼠心肌MMP-2、Ⅲ型胶原和JNK1／2表达的影响

目的研究Ⅲ型胶原、基质金属蛋白酶-2(matrix metalloproteinases-2，MMP-2)和JNK1／2在高血压大鼠心肌组织中的表达及氯沙坦干预后3者的变化，探讨氯沙坦逆转心室重构的药理机制。方法采用SD大鼠建立两肾一夹型(Goldblatt)高血压模型，将大鼠随机分为对照组、高血压组和氯沙坦治疗组。采用鼠尾动脉测压法记录每周血压变化，实验末切取心脏，并计算心脏与体重比值。免疫组织化学方法测定心脏组织中的Ⅲ型胶原、基质金属蛋白酶-2和JNK1／2表达。结果左肾动脉结扎术后大鼠收缩压升高，心脏体重比值增加，心肌间质Ⅲ型胶原、基质金属蛋白酶-2、心肌细胞JNK1／2表达增多。氯沙坦治疗能显著降低大鼠收缩压，降低心脏体重比值，并且使心肌间质Ⅲ型胶原、基质金属蛋白酶-2、心肌细胞JNK1／2表达减少。结论两肾一夹型高血压大鼠重构心肌中MMP-2、Ⅲ型胶原、JNK1／2表达升高。氯沙坦能有效逆转心肌肥厚或心室重构，这一效应与其降低心肌中高表达的MMP-2，Ⅲ型胶原、JNK1／2有关。     

基质金属蛋白酶-2在高血压大鼠心脏中表达的意义及氨氯地平对其的影响

目的:探讨基质金属蛋白酶-2(MMP-2)在脱氧皮质酮-盐型高血压大鼠(DHR)心脏微小血管重塑中作用及其可能的调节机制。方法:30只雄性SD大鼠,切除左侧肾脏后1周,随机等分为3组。对照组,饮自来水;另2组分别为模型组和氨氯地平组,开始每周予脱氧皮质酮50 mg.kg-1,ip,连续5周;氨氯地平组用氨氯地平30 mg.kg-1.d-1灌胃,连续5周。5周末处死动物,按相应的要求留取血液和心脏标本,分别行血浆内皮素-1(ET-1)浓度、微小血管密度及MMP-2/MMP-2特异性组织抑制因子(TIMP-2)蛋白和基因表达的检测,评价在氨氯地平干预下微小血管病变与MMP-2/TIMP-2表达间关系。结果:在DHR左心室心内膜下心肌中存在微小动脉密度增加和毛细血管密度减少,MMP-2的mRNA和MMP-2/TIMP-2的蛋白表达上调;氨氯地平能抑制血压升高,明显减轻微小血管损害,下调MMP-2/TIMP-2的mRNA和蛋白表达。MMP-2的表达同微小血管密度间具有良好的相关性。结论:在DHR心脏中存在微小血管病变,MMP-2/TIMP-2表达可能参与微小血管病变的病理机制,血压可能是通过调节MMP-2/TIMP-2表达参与微小血管病变;干预MMP-2/TIMP-2的表达可能为防治高血压靶器官损害的新的靶点。     

2型糖尿病大鼠心肌组织MMP-2、MMP-9、TIMP-1的表达与胶原代谢的关系

目的 了解2型糖尿病（T2DM）大鼠模型心肌组织基质金属蛋白酶2（MMP-2）、基质金属蛋白酶9（MMP-9）及其组织抑制物1（TIMP-1）的mRNA表达变化、蛋白水平及组织定位，探讨MMPs／TIMPs在T2DM心肌病变发生发展中的作用。方法 Masson染色观察心肌胶原含量变化；RT-PCR方法观察心肌MMP-、MMP-9和TIMP-1mRNA表达；免疫组化方法测定MMP-2、MMP-9、TIMP-1的蛋白表达和组织定位。结果 T2DM大鼠心肌胶原纤维明显增多；MMP-2mRNA表达明显下调，蛋白水平下降；MMP-9、TIMP-1的mRNA表达均增加，MMP-9、TIMP-1的蛋白水平也升高，但MMP-9／TIMP-1的比值下降。结论 T2DM大鼠心肌组织胶原聚集，心肌纤维化。MMPs／TIMP-1比例失衡可能与T2DM心肌病变的发生有关。     

肾上腺髓质素缺失对单侧输尿管结扎小鼠MMP-1/TIMP-1表达的影响

目的 探讨肾上腺髓质素在肾间质纤维化实验动物肾脏基质金属蛋白酶-1/基质金属蛋白酶抑制剂-1(MMP-1/TIMP-1)表达中的作用.方法 结扎肾上腺髓质素基因敲除小鼠及野生小鼠单侧输尿管,制备肾间质纤维化实验动物模型并给予缬沙坦灌胃治疗,采用免疫组化、原位杂交、RT-PCR方法检测MMP-1及TIMP-1的表达.结果 单侧输尿管结扎 (UUO) 小鼠肾脏MMP-1的表达减弱,而TIMP-1的表达显著上调,基因敲除小鼠较野生小鼠更为明显,采用缬沙坦治疗可部分抑制TIMP-1的表达.结论 肾上腺髓质素可以通过纠正MMP-1/TIMP-1的失衡,对UUO介导的肾间质纤维化起到保护作用.     

硫化氢通过调控转化生长因子-β及基质金属蛋白酶-9/基质金属蛋白酶抑制剂-1改善糖尿病大鼠肾小管间质纤维化

目的探讨硫化氢(H_2S)气体信号分子对糖尿病大鼠肾小管间质纤维化及转化生长因子(TGF)-β、基质金属蛋白酶(MMP)-9/基质金属蛋白酶抑制剂(TIMP)-1表达的影响。方法单次腹腔注射链脲佐菌素(STZ)建立糖尿病模型将成年雄性SD大鼠64只随机分为正常对照组(Control组)、STZ组、STZ+H_2S组、H_2S组。造模72 h后采尾静脉血检测,若血糖>16.7 mmol/L表明造模成功。H_2S组和STZ+H_2S组大鼠腹腔注射Na HS溶液100μmol·kg~(-1)·d~(-1),Control组和STZ组腹腔注射同等量生理盐水。8 w后取标本,Masson染色观察大鼠肾脏组织病理形态学改变,Western印迹检测大鼠肾脏TGF-β、MMP-9、TIMP-1、Ⅳ型胶原蛋白的表达水平。结果与Control组相比,STZ组存在明显肾小管间质纤维化,同时肾脏组织中Ⅳ型胶原、TGF-β、TIMP-1蛋白的表达升高,MMP-9蛋白的表达水平下降(P<0.05);与STZ组大鼠相比,STZ+H_2S组肾小管间质纤维化减轻,肾组织TGF-β、TIMP-1、Ⅳ型胶原蛋白的表达降低,MMP-9蛋白表达水平升高(P<0.05)。结论 H_2S可改善糖尿病大鼠肾小管间质纤维化,其机制可能与调控MMP-9/TIMP-1失衡以及TGF-β蛋白的表达水平有关。     

Focal and segmental glomerular sclerosis in reflux nephropathy

Reflux nephropathy was diagnosed in 23 patients (14 per cent of all the patients who received transplants) between 1973 and 1977, and nephrectomy was performed in all. Histology and immunofluorescence revealed a glomerular sclerosis associated with the idiopathic nephrotic syndrome. No focal and segmental glomerular sclerosis was seen in kidneys removed from patients with nonglomerular renal disease. Twenty-four hour urinary protein excretion in grams was 3.1 ± 0.3 (mean ± SEM) and was greater than that in our patients with end-stage nonglomerular renal disease. Thirty-one renal transplants were performed in these 23 patients; thereafter, maximum protein excretion was 1.4 g. Focal and segmental glomerular sclerosis was seen in only one (chronic rejection, protein excretion < 0.5) of the 20 kidneys available for histologic study. Thus, focal and segmental glomerular sclerosis is extremely common in reflux nephropathy, accounts for “glomerular” proteinuria and may contribute importantly to progressive renal failure but, unlike that associated with the idiopathic nephrotic syndrome, rarely recurs after renal transplantation.     

19例家族性局灶节段性肾小球硬化的临床研究

目的:通过对家族性局灶节段性肾小球硬化(FSGS)患者家系调查和总结,探讨家族性FSGS特点. 方法:前瞻性对2006-01至2007-12入住我科并经肾活检确诊的非继发性FSGS患者进行家系调查,对其中有家族史的FSGS患者进行家系成员筛查,同时行IV型胶原和α半乳糖酐酶活性检查排除Alport综合征和Fabry病,比较家族性FSGS和散发性FSGS患者临床特点. 结果:我院2006年1月～2007年12月经肾活检确诊的非继发性FSGS共213例,其中家族性FSGS共19例,占非继发性FSGS总病例的8.9%.19例家族性FSGS中男性10例,女性9例,和散发性FSGS性别相比(男∶女＝100∶94)无显著差异(P>0.05);家族性FSGS患者平均24h尿蛋白水平(1.6±1.6)g,与散发性FSGS患者(1.8±1.7)g相比无显著差异(P>0.05);家族性FSGS先证者中血肌酐(167.5±166.6) μmol/L,与散发性FSGS (198.4±190.6) μmol/L相比无显著差异(P>0.05).19个家系中有5个家系各有2例患者经肾活检确诊为FSGS,7个家系为1例肾活检证实FSGS伴1例或1例以上ESRF,7个家系为1例肾活检证实FSGS伴有1例或1例以上亲属蛋白尿,或不明原因的肾功能不全. 结论:家族性FSGS发生率较高,达非继发性FSGS总数的8.9%,临床表现方面家族性和散发性FSGS相比无明显差异,较易漏误诊,通过家系调查可以明显提高家族性FSGS诊断率.重视家族性FSGS的筛查,也将有助于遗传性肾脏疾病的进一步研究.     

The prognosis of focal segmental glomerular sclerosis of adulthood

We describe 46 adults with idiopathic focal segmental glomerular sclerosis (FSGS). The mean age was 36.9 years (range, 15 to 80 years). Males represented 61%, and 65.2% were white. Hypertension was a presenting feature in 63% and 32.6% had microscopic hematuria. Twenty-nine patients had nephrotic proteinuria (greater than or equal to 3.0 g/24 h) at presentation, and 13 had renal insufficiency (serum creatinine concentration greater than 1.5 mg/dl). A mean follow-up of 59.8 months (range, 3 to 255 months) was obtained. In addition to segmental sclerosis, glomerular hyalinosis was observed in 65.3% of biopsies, and this was similar irrespective of the severity of proteinuria. Sixteen of the 29 patients with nephrotic proteinuria received prednisone therapy (60 mg/day) for at least 1 month. Three received cytotoxic agents in addition. A response to therapy was observed in 50%, 5 achieving a complete remission and 3 a partial remission. No patient with non-nephrotic proteinuria received prednisone therapy. The clinical course of each patient was evaluated based on the slope calculated by the linear regression method using the inverse of serum creatinine from the time of presentation to follow-up. Patients with non-nephrotic proteinuria had a better prognosis than nephrotics (P less than .05). Nephrotic patients responding to therapy had a better course than non-responders or patients not treated (P less than 0.01). At the time of last follow-up, 8 patients had progressed to end-stage renal disease, 6 of whom had presented with nephrotic proteinuria. No patient responding to therapy had progressed to end-stage renal disease.(ABSTRACT TRUNCATED AT 250 WORDS)     

乙型肝炎病毒引起局灶节段性肾小球硬化1例

1病历资料患者女,33岁。于2002年7月感冒后出现颜面及双下肢水肿、肉眼血尿,无发热、寒战、尿频、尿急、尿痛,无潮热、盗汗、咯血、胸痛、呼吸困难,无心悸、心累、端坐呼吸及夜间呼吸困难,无光过敏、脱发、口腔溃疡、关节红肿疼痛、皮肤黏膜瘀斑,无腹痛、腹泻、解黑大便等。在当地医院就诊,查     

Focal glomerular sclerosis: contrasting clinical patterns in children and adults.

In a retrospective clinicopathological study, 48 kidney biopsy specimens from 16 children (mean age, 7 years) and 17 adults (mean age, 33 years) with histological evidence of focal glomerular sclerosis (FGS) were examined using light, immunofluorescence and electron microscopy. The histopathological findings were related to the clinical course of each patient. At the clinical onset of the disease, the nephrotic syndrome was seen more commonly in children (12/16) than adults (7/17), while the incidence of both hypertension (children 1/16 versus adults, 9/17) and renal insufficiency (children, 0/16 versus adults, 7/17) was greater in adults. Despite a shorter average follow-up, (adults 3 10/12 years versus children, 7 years), the incidence of hypertension (adults, 13/17 versus children, 7/16) and renal functional impairment (adults, 13/17 versus children, 3/16) remained greater in the adult patients. One child and three adults died in renal failure while two adults underwent transplantation and on requires regular dialysis therapy. Nine of 15 pediatric patients treated with corticosteroids experienced partial or complete remission in either their nephrotic syndrome or level of urine protein excretion, while just 3 of 6 adult patients treated with corticosteroids experienced a partial remission, but never became protein-free. There was an excellent correlation in all patients between the degree of functional renal impairment and the extent of glomerular and nonglomerular histopathological damage in the kidney. It is concluded that in the adults, FGS represents a more severe and progressive disease process and is less responsive to therapy.     

Advanced glycation end products induce glomerular sclerosis and albuminuria in normal rats.

High levels of tissue advanced glycation end products (AGEs) that result from the spontaneous modification of proteins by glucose occur in diabetes and aging. To address the potential pathogenic role of AGEs in the glomerulosclerosis of diabetes or nephrosclerosis of aging, doses of AGE-modified rat albumin (25 mg per kg per day, i.v.) sufficient to elevate circulating AGE levels to the range of diabetic serum were administered daily to healthy rats alone or in combination with the AGE inhibitor aminoguanidine. After 5 months, the AGE content of renal tissues in AGE-treated rats rose to 50% above controls (P < 0.025), whereas serum contained 2.8-fold greater AGE levels (P < 0.025). Light and electron microscopy of kidneys from AGE-treated rats revealed a more than 50% increase in glomerular volume compared to controls (P < 0.001), significant periodic acid/Schiff reagent-positive deposits, basement membrane widening, and mesangial extracellular matrix increase and indicated significant glomerulosclerosis compared to untreated (P < 0.002) or albumin-treated controls (P < 0.002). These changes were associated with significant loss of protein (P < 0.005) and albumin (P < 0.002) in the urine of AGE-treated rats compared to controls. Cotreatment with aminoguanidine markedly limited both the structural and functional defects. These in vivo data demonstrate that AGEs influence glomerular structure and function in a manner leading to glomerulosclerosis. The effects are AGE-specific, as they are ameliorated by a pharmacological AGE inhibitor, aminoguanidine.     

Mice with focal segmental glomerular sclerosis are induced by adriamycin and its mechanism

<正>Objective To establish adriamycin-induced focal segmental glomerular sclerosis(FSGS)mice model,and observe the expressions of and relation between oxidative stress and p38 MAPK signal pathway in renal injury.Methods Eight-week-old male Balb/c mice were randomly divided into FSGS group(n=20)and control     

足细胞特异性因子nephrin及肿瘤蛋白-1在肾小球硬化过程中表达变化

目的：应用嘌罗霉素氨基核苷（PAN）模型，通过观察临床、病理指标及足细胞特异性因子nephrin和肿瘤蛋白-1（WT-1）在足细胞的表达，了解足细胞损害在肾小球硬化过程中的重要作用。方法尾静脉注射PAN制作大鼠肾病模型，测定不同时间点的生化指标，处死大鼠，取其肾脏，石蜡包埋切片，PAS及免疫组化染色，测定肾小球硬化指数（GSI），肾小球面积（GA），nephrin及WT-1的表达，进行统计学分析。结果 PAN组在早期（第8天）表现为典型肾病综合征，后期（20周）表现为慢性肾炎。形态学方面，PAN组的GSI在4，14，20周均明显高于对照组（P＜0.05）。PAN组GA在14周时大于对照组（P＜0.01），但在20周时小于对照组（P＜0.001），并且20周时GSI与GA之间呈强负相关关系。PAN组nephrin表达在4，14，20周时与对照组比较均有所减少（P＜0.05），与此同时，WT-1的表达在14和20周时较对照组明显减少（P＜0.001）。结论（1）在肾炎发展中，随着时间延长，GSI增高，GA缩小，并且GSI和GA之间呈负相关。（2）在肾小球硬化发展中， nephrin和WT-1的表达进行性减少，说明有明显足细胞数量的减少。因此，足细胞的损伤与肾小球硬化密切相关。