结缔组织生长因子在输尿管梗阻大鼠肾间质中的表达

笔者以单侧输尿管结扎建立梗阻性肾病大鼠(UUO)的肾间质纤维化模型,检测转化生长因子-β(TGF-β)、结缔组织生长因子(CTGF)在肾间质中的表达,同时检测细胞外基质中纤维连结蛋白(FN)和平滑肌肌动蛋白(α-SMA)的表达。     

结缔组织生长因子mRNA表达在肾间质纤维化的作用

肾间质纤维化是多种肾脏疾病走向肾功能衰竭的共同病理过程.其发病机制与细胞因子表达、细胞凋亡等多因素相关。结缔组织生长因子（CTGF）mRNA在大鼠肾间质纤维化中作用相关研究已成为一个研究热点。本研究旨在通过观察CTGFmRNA的分布与表达,为CTGF在肾间质纤维化发病机制中的关键作用提供一定的证据。     

结缔组织生长因子在肾纤维化中的作用

肾纤维化(renal fibrosis)是所有肾脏疾病进展至终末期肾衰竭的必经之路.它主要表现为肾间质纤维化(renalinterstitialfibrosis)及肾小球硬化(gromerulosclerosis)两方面.主要特征为肾小管和间质毛细血管的丧失及细胞外基质(extracellular matrix,ECM)的过度积聚.     

结缔组织生长因子与肾纤维化的研究进展

作为TGF-β的下游效应因子,结缔组织生长因子因其具有多种生物学活性,尤其是在纤维化过程中的显著作用,引起人们越来越多的关注。本文就结缔组织生长因子的结构、生物学活性及其在肾纤维化过程中的作用进行综述。     

结缔组织生长因子在关木通相关肾小管间质肾病肾间质纤维化中的作用

目的 通过观察关木通相关肾小管间质肾病(GMT-TIN)肾小管间质内纤维化相关生长因子表达及细胞外基质(ECM)沉积的特点,探讨其纤维化病变发生与发展的有关环节。方法 22例GMT-TIN患者根据病理特点分为急性组8例、慢性轻型组8例和慢性重型组6例,对其纤维化程度进行半定量分析。应用免疫组化SP法观察肾活检组织标本中,肾小管间质内α-平滑肌肌动蛋白(α-SMA)、纤连蛋白(FN)、IV型胶原、结缔组织生长因子(CTGF)和转化生长因子 β1(TGF-β1)的表达情况。分析上述指标之间以及与纤维化病变之间的相关关系。结果 (1)各组肾小管间质内均检测到高表达的α-SMA、FN和IV型胶原,随病变慢性化加重3者表达均明显增强,并与肾间质纤维化程度正相关(P<0.01)。(2)各组肾间质内均出现CTGF及TGF-β1表达,2者间显著正相关(r=O.771,P<0.0001),但在时相上存在一定差异。(3)肾间质内CTGF表达分别与FN及IV型胶原沉积正相关(r=0.6855和0.5964,P<0.01)、与纤维化病变程度正相关(r=0.4941,P<0.05)。结论(1)GMT-TIN肾间质内CTGF的高表达、CTGF与ECM沉积以及纤维化病变的相关性,提示其在GMT-TIN肾间质纤维化病变中起重要作用。(2)CTGF与TGF-β1的相关关系及2者表达时相的差异,进一步支持CTGF可能作为TGF-β1的下游效应因子在纤维化病变     

肿瘤细胞上清液对成纤维细胞表型转变及血管内皮生长因子-A表达的影响

目的检测肿瘤细胞上清液对成纤维细胞的激活情况及激活后血管内皮生长因子-A(VEGF-A)表达的变化规律。方法 MTT法检测普通培养液、含转化生长因子-β1(TGF-β1)的普通培养液以及肿瘤细胞上清液组成的条件培养液对成纤维细胞生长情况的影响,用RT-PCR、Western blot及免疫组织化学法检测不同培养条件下成纤维细胞的α-平滑肌肌动蛋白(α-SMA)与VEGF-A的表达。结果肿瘤细胞上清液对成纤维细胞的生长有一定的促进作用。含TGF-β1的普通培养液比普通培养液更有利于成纤维细胞转化为肌成纤维细胞,并且能维持肌成纤维细胞的表型,但是二者均不表达VEGF-A;条件培养液能促进成纤维细胞稳定表达α-SMA和VEGF-A,二者在培养后第1天均开始表达,第3天表达量达到峰值,第3天以后表达稳定。结论肿瘤细胞上清液能够有效、稳定地激活成纤维细胞为肌成纤维细胞,成纤维细胞的激活程度影响VEGF-A的表达,二者均具有最佳的激活时间点,并且最佳时间点相一致,最佳激活点的成纤维细胞有望成为一种可用于移植的促进血管重建的细胞。     

黄芪当归合剂及依那普利对结缔组织生长因子在肾间质纤维化中表达的比较研究

目的 探讨结缔组织生长因子(CTGF)在肾间质纤维化中的表达及其在依那普利、黄芪当归合剂的肾脏保护作用中的意义。方法 采用单侧输尿管结扎致肾间质纤维化大鼠模型。将大鼠随机分为4组:假手术组、模型组、黄芪当归组、依那普利组。术后第9天处死各组大鼠,经免疫组织化学(组化)、Western蛋白印迹分析方法观察各组CTGF的表达部位及蛋白表达水平。用免疫组化半定量检测各组肾间质的α-平滑肌肌动蛋白(α-SMA)的表达。用天狼星红染色半定量检测以Ⅰ型胶原为主的肾间质胶原成份。Masson染色评定各组肾小管间质损害程度。结果 模型组CTGF、α-SMA、Ⅰ型胶原的表达及肾小管间质损伤指数明显高于假手术组(P<0.01),而黄芪当归组及依那普利组明显低于模型组(P<0.01)。两治疗组间比较,除结缔组织生长因子外(P<0.01),其余指标无显著性差异(P>0.05)。各项指标作相关分析,CTGF与肾小管间质损伤指数(r=0.788,P<0.01)、α-SMA(r=0.940,P<0.01)、Ⅰ型胶原(r=0.820,P<0.01)为正相关关系。结论 黄氏当归合剂及依那普利可能通过下调CTGF而减轻肾间质纤维化。     

结缔组织生长因子与肾脏纤维化研究进展

近年来，结缔组织生长因子在肾脏疾病中的作用倍受关注。本文对结缔组织生长因子的结构、生物学功能及其作用机制作一介绍，并综述了近年来国外关于结缔组织生长因子在肾脏中的表达以及它在肾脏纤维化中作用机制方面的研究进展。     

缺氧诱导因子-1α在肾间质纤维化中的表达及意义

目的：探讨缺氧诱导因子-1α（HIF-1α）致肾间质纤维化的作用。方法：60只雄性SD大鼠随机分为假手术组（SOR）和单侧输尿管梗阻（UUO）模型组。术后第3天,第7天,第14天各处死大鼠10只,肾组织行HE染色并观察病理变化。采用免疫组织化学和荧光定量逆转录聚合酶链反应（Q-RT-PCR）法检测肾脏组织中HIF-1α、CTGF、TGF-β1蛋白和mRNA表达。结果：与假手术组相比,模型组大鼠肾脏病理损害进行性加重;HIF-1α、CTGF、TGF-β1蛋白表达随梗阻时间的延长逐渐增加;与假手术组相比HIF-lα、CTGF、TGF-β1mRNA表达明显增加（P〈0.05）;相关分析显示,模型组第3天HIF-lαmRNA表达与CTGFmRNA,TGF-β1mRNA表达分别呈正相关（r=0.748,0.659,P〈0.05）,模型组第7天组HIF-1αmRNA分别和CTGFmRNA、TGF-β1mRNA呈正相关（分别为r=0.663,0.645,P〈0.05）,模型组第14天组HIF-1αmRNA分别和CTGFmRNA、TGF-β1mRNA呈正相关（分别为r=0.515,0.752,P〈0.05）。结论：HIF-1α可能通过调节CTGF、TGF-β1的表达参与了肾间质纤维化发生和发展的过程。     

结缔组织生长因子与肾脏纤维化研究进展

近年来 ,结缔组织生长因子在肾脏疾病中的作用倍受关注。本文对结缔组织生长因子的结构、生物学功能及其作用机制作一介绍 ,并综述了近年来国外关于结缔组织生长因子在肾脏中的表达以及它在肾脏纤维化中作用机制方面的研究进展。     

结缔组织生长因子在结直肠癌中的表达及临床意义

目的 探讨结缔组织生长因子(connective tissue growth factor,CTGF)在结直肠癌组织中的表达及临床意义.方法 采用免疫组化法检测62例结直肠癌及其对应的癌旁组织中CTGF的表达,并对结果进行统计学分析.结果 CTGF在结直肠癌组织和癌旁组织中表达阳性率分别为61.3%(38/62)和19.4%(12/62),其在结直肠癌组织中的表达明显高于其癌旁组织(P＜0.05).CTGF在结直肠癌组织中的表达与其分化程度、浸润深度及淋巴结转移有关,即癌组织分化程度越低,CTGF表达阳性率越低(P=0.030),随癌组织浸润深度的加深而CTGF表达阳性率明显降低(P=0.032),有淋巴结转移者CTGF的表达阳性率明显低于无淋巴结转移者(P=0.017),而与性别无关(P>0.05).结论 CTGF在结直肠癌的发生、发展过程中可能起重要作用,对指导临床治疗和判断预后有重要意义.     

转化生长因子-β对人肾小球系膜细胞结缔组织生长因子及细胞外基质成分表达的影响

目的:探讨转化生长因子-β(TGF-β)对人肾小球系膜细胞(HMCs)结缔组织生长因子(CTGF)及细胞外基质(ECM)成分表达的影响.方法:应用逆转录-聚合酶链反应(RT-PCR)和蛋白印迹技术,观察不同时间TGF-β1对HMCs CTGF mRNA及其蛋白质表达的影响,同时观察对Ⅰ型胶原、Ⅳ型胶原和纤维结合蛋白(FN)mRNA表达的影响.结果:在正常培养情况下,HMCs有CTGF mRNA及其蛋白质的表达.HMCs在TGF-β1刺激后,CTGFmRNA表达明显增加,6 h达到高峰,可持续增高到24 h;CTGF蛋白表达12 h达到高峰,可持续增高至24 h.Ⅰ型胶原、Ⅳ型胶原mRNA表达逐渐增加,24 h明显增加并达到高峰.FN mRNA12 h达到高峰,24 h后开始回落.结论:HMCs可表达CTGF.TGF-β可诱导体外培养的HMCsCTGF及ECM成分高表达.     

Connective Tissue Growth Factor Gene Expression Alters Tumor Progression in Esophageal Cancer

The ability of cancer cells to initiate specific fibroblast reactions may subsequently determine tumor evolution. In the present study we examined the coordinated expression of transforming growth factor-beta-1 (TGF-b1), its signaling receptors, and its downstream mediator—connective tissue growth factor (CTGF)—and their impact on tumor progression and fibrogenesis in esophageal carcinomas. Messenger ribonucleic acid (mRNA) expression of TGF-b1, CTGF, TGF-b receptor subtype I ALK5 (TbR-IALK5), and TGF-b receptor type II (TbR-II) was studied by Northern blot analysis in esophageal cancer and the normal esophagus. By means of immunohistochemistry and Western blot analysis, the respective proteins were localized in the tissue samples and the protein content was quantitated. Northern blot analysis revealed 3-fold and 4-fold increases (p <0.05) in TGF-b1 and CTGF mRNA levels, respectively, in esophageal cancer in comparison with normal controls, whereas TbR-I mRNA levels were significantly decreased and TbR-II mRNA levels were unchanged in the cancer samples. Immunostaining revealed results similar to those seen on the RNA level. TGF-b1 and CTGF immunoreactivity were increased, TbR-II was unchanged, and TbR-IALK5 immunoreactivity was decreased. CTGF immunoreactivity was mainly present in the stroma surrounding the cancer cells but was also present in the cancer cells. The degree of fibrosis was different in squamous and adenocarcinomas and was significantly related to CTGF mRNA expression levels. The presence of CTGF in squamous cell carcinomas was associated with longer survival, whereas in adenocarcinomas it influenced survival negatively. The findings indicate that TGF-b signaling is disturbed in esophageal cancer. CTGF, a downstream effector of TGF-b action, differentially influences the composition of tumor microenvironment and distinct cell-matrix interactions in the two histological types of esophageal carcinoma, resulting in differences in tumor progression and patient survival.     

Connective Tissue Growth Factor Is a Regulator for Fibrosis in Human Chronic Pancreatitis

OBJECTIVE: To evaluate the parameters that mediate fibrogenesis in chronic pancreatitis (CP). BACKGROUND: Connective tissue growth factor (CTGF), which is regulated by transforming growth factor beta (TGF-beta), has recently been implicated in skin fibrosis and atherosclerosis. In the present study, the authors analyzed the concomitant presence of TGF-beta1 and its signaling receptors-TGF-beta receptor I, subtype ALK5 (TbetaR-I(ALK5)), and TGF-beta receptor II (TbetaR-II)-as well as CTGF and collagen type I in the pancreatic tissue of patients undergoing surgery for chronic pancreatitis. PATIENTS AND METHODS: CP tissue samples were obtained from 40 patients (8 women, 32 men) undergoing pancreatic resection. Tissue samples of 25 previously healthy organ donors (12 women, 13 men) served as controls. The expression of TGF-beta1, TbetaR-I(ALK5), TbetaR-II, CTGF, and collagen type I was studied by Northern blot analysis. By in situ hybridization and immunohistochemistry, the respective mRNA moieties and proteins were localized in the tissue samples. RESULTS: Northern blot analysis showed that CP tissue samples exhibited concomitant enhanced mRNA expression of TGF-beta1 (38-fold), TbetaR-II (5-fold), CTGF (25-fold), and collagen type I (24-fold) compared with normal controls. In addition, TbetaR-I(ALK5) mRNA was increased in 50% of CP tissue samples (1.8-fold). By in situ hybridization, TGF-beta1, TbetaR-I(ALK5), and TbetaR-II mRNA were often seen to be colocalized, especially in the ductal cells and in metaplastic areas where atrophic acinar cells appeared to dedifferentiate into ductal structures. In contrast, CTGF was located in degenerating acinar cells and principally in fibroblasts surrounding these areas. Moreover, CTGF mRNA expression levels correlated positively with the degree of fibrosis in CP tissues. CONCLUSION: The concomitant overexpression of CTGF, collagen type I, TGF-beta1, and its signaling receptors in CP suggests that these proteins contribute to enhanced extracellular matrix synthesis and accumulation, resulting finally in the fibrogenesis observed in CP.     

绞股蓝总皂苷防治单侧输尿管结扎大鼠肾间质纤维化的实验研究

目的探讨绞股蓝总皂苷(GPs)对单侧输尿管结扎(UUO)大鼠肾组织结缔组织生长因子(CTGF)表达及肾间质纤维化的影响。方法采用UUO大鼠模型,将大鼠随机分为3组假手术组、模型组、GPs组。假手术组和模型组仅给予标准饲料30g,GPs组于术前3d至术后9d每天给予GPs200mg·kg-1·d-1灌胃,第9d处死各组大鼠。免疫组化法检测各组肾组织CTGF、转换生长因子β1(TGF-β1)和α-平滑肌肌动蛋白(α-SMA)的表达;RT-PCR方法检测各组CTGFmRNA含量;Masson染色评定各组肾小管间质损害程度。结果模型组CTGF、TGF-β1、α-SMA的表达及肾小管间质损伤指数明显高于假手术组(P<0.01),而GPs组各项指标明显低于模型组(P<0.01)。各项指标作相关分析,CTGF与肾小管间质损伤指数(r=0.788,P<0.01)、TGF-β1(r=0.879,P<0.01)、α-SMA(r=0.940,P<0.01)为正相关关系。结论绞股蓝总皂苷可以抑制肾纤维化时结缔组织生长因子表达,从而遏制肾纤维化的进展。     

结缔组织生长因子在BPH患者膀胱壁中的表达及意义

目的：探讨结缔组织生长因子（CTGF）在BPH患者膀胱壁中的表达,评价CTGF的表达和膀胱出口梗阻所致的膀胱壁纤维化之间的关系。方法：分别留取BPH患者膀胱壁标本25例和对照组膀胱壁标本19例,通过逆转录-聚合酶链反应（RT—PCR）和图像半定量分析,检测组织中CTGF基因表达水平。结果：前列腺增生症和对照组患者膀胱壁中CTGF mRNA指数分别为2.00±0．74、0,79±0．15,两组相比较差异显著,有统计学意义t=8．24（P〈0．01）。结论：CTGF在前列腺增生症患者的膀胱壁组织中表达明显增高,可能是其纤维化的重要促进因素。     

积雪草颗粒对单侧输尿管结扎大鼠肾组织结缔组织生长因子表达的影响

目的：研究积雪草颗粒（CTA）对单侧输尿管结扎（UUO）大鼠肾组织结缔组织生长因子（CTGF）表达的影响,为临床上应用CTA延缓肾衰竭的进展提供理论依据。方法：采用UUO大鼠模型,60只雄性SD大鼠随机分为6组：假手术（SOR）组、模型（UUO）组、积雪草颗粒大剂量组（CTAh）、积雪草颗粒中剂量组（CTAm）、积雪草颗粒小剂量组（CTAl）、蒙诺组（Monopril）。术后SOR、UUO组管饲与药物组同等容积的生理盐水;CTAh、CTAm、CTAl组于术后即每日分别管饲积雪草颗粒11.2mg.100g-1.d-1、5.6mg.100g-1.d-1、2.8mg.100g-1.d-1;Monopril组每日管饲蒙诺20mg.100g-1.d-1,术后21d处死各组大鼠。免疫组化法检测各肾组织CTGF的表达,RT-PCR方法检测各组CTGF mRNA含量。结果：UUO组CTGF的表达明显高于SOR组（P〈0.01）,而CTAh组和Monopril组各项指标明显低于UUO组（P〈0.01）,CTAh组和Monopril组无统计学差异（P〉0.05）,CTA组间比较大剂量组抑制CTGF表达优于中、小剂量组（P〈0.01）。结论：CTA减轻RIF的程度可能与下调CTGF的表达有关,且随剂量增加其抑制CTGF表达的生物学效应亦增大,呈量效依赖关系,CTA大剂量组抑制CTGF的效应与蒙诺组相似。     

Alteration of Epidermal Growth Factor Receptor Expression Following Ischaemia of Renal Tissue

This study was aimed to investigate Epidermal Growth Factor Receptor (EGF-R) expression after ischaemic injury in renal tissue and the effects of calcium channel blockers in the prevention of damage due to ischaemic insult. Simple nephrectomy was performed in a group of Sprague-Dawley rats, and kidneys were grouped according to cold ischaemia time (1, 6, 12, 24 and 48 hours, respectively) and to the type of calcium channel blockers (diltiazem and verapamil) used. EGF-R expression status was investigated in each group by immunohistochemistry on paraffin sections. Overall expression of EGF-receptor was detected in 8 822.8%) kidneys. In terms of localization of EGF-receptor expression cortical tubular staining was detected in 8 (100%) kidneys, medullar tubular staining in (62.5%) kidneys and glomerular mesangial staining in 5 (62.5%) kidneys. There was no difference between various ischaemia times and different calcium channel blockers used. It has been concluded that hypoxia and cold ischaemia causes widespread down-regulation of EGF-receptor expression in renal tissue regardless of treatment with calcium channel blockers.     

结缔组织生长因子(CTGF)在前列腺疾病中的研究

结缔组织生长因子(CTGF)是CCN家族的成员之一,在成年人的心脏、脑、胎盘、肺、肝脏、肌肉、肾脏等多个脏器的组织中均有所表达,具有促进有丝分裂、黏附、细胞凋亡,细胞外基质沉积以及细胞迁移等作用.随着研究的深入,发现其在前列腺疾病的发生发展中也发挥了重要作用,可能成为治疗前列腺疾病的新靶点.本文就结缔组织生长因子在前列腺疾病中的研究作一综述.     

Connective Tissue Growth Factor is Involved in Pancreatic Repair and Tissue Remodeling in Human and Rat Acute Necrotizing Pancreatitis

OBJECTIVE: To analyze the involvement of connective tissue growth factor (CTGF) in the transforming growth factor-beta (TGF-beta) pathway during acute necrotizing pancreatitis (ANP) in humans and rats. SUMMARY BACKGROUND DATA: Connective tissue growth factor is involved in several fibrotic diseases and has a critical role in fibrogenesis and tissue remodeling after injury. METHODS: Normal human pancreas tissue samples were obtained through an organ donor program from five individuals without a history of pancreatic disease. Human ANP tissues were obtained from eight persons undergoing surgery for this disease. In rats, ANP was induced by intraductal infusion of taurocholate. The expression of CTGF was studied by Northern blot analysis, in situ hybridization, and immunohistochemistry in both human and rat pancreatic tissue samples. RESULTS: Northern blot analysis revealed enhanced CTGF mRNA expression in human ANP tissue samples compared with normal controls. In addition, a concomitant increase in TGF-beta1 was present. By in situ hybridization, CTGF mRNA was localized in the remaining acinar and ductal cells and in fibroblasts. In regions of intense damage adjacent to areas of necrosis, CTGF mRNA signals were most intense. Inflammatory cells were devoid of any CTGF mRNA signals. By immunohistochemistry, CTGF protein was localized at high levels in the same cell types as CTGF mRNA. In ANP in rats, concomitantly enhanced mRNA levels of CTGF, TGF-beta1, and collagen type 1 were present, with a biphasic peak pattern on days 2 to 3 and day 7 after induction of ANP. CONCLUSIONS: These data indicate that CTGF participates in tissue remodeling in ANP. The expression of CTGF predominantly in the remaining acinar and ductal cells indicates that extracellular matrix synthesis after necrosis is at least partly regulated by the remaining pancreatic parenchyma and only to a minor extent by inflammatory cells. Blockage of CTGF, a downstream mediator of TGF-beta in fibrogenesis, might be useful as a target to influence and reduce fibrogenesis in this disorder.