Bone Morphogenetic Protein-7 and Connective Tissue Growth Factor: Novel Targets for Treatment of Renal Fibrosis?

Renal fibrosis is the major determinant in progression of kidney disease and results from an inappropriate response to acute and chronic kidney injury. Transforming growth factor (TGF)-β1 is the driving force behind renal fibrosis and has since long been regarded as the key factor to be targeted in prevention and treatment of renal fibrosis. Despite the impressive results obtained in experimental renal fibrosis, TGF-β1 blockade has not yet translated into an effective and safe therapeutic in human patients. Therefore, it remains important to explore the role of additional growth factors which are involved in renal regeneration and fibrosis. Recently, bone morphogenetic protein (BMP)-7 and connective tissue growth factor (CTGF) have both emerged as novel modulators of profibrotic TGF-β1 activity. The expression of BMP-7 is decreased in various models of renal disease, while CTGF is strongly upregulated in experimental and human renal fibrosis. In experimental kidney injury, administration of BMP-7 or inhibition of CTGF have been sufficient to result in striking improvement of renal function and structure. This review summarizes the current knowledge of BMP-7 and CTGF in the kidney, and discusses their therapeutic potential in renal fibrosis.     

CTGF siRNA质粒对UUO小鼠肾脏间质纤维化的影响

目的利用单侧输尿管梗阻(UUO)小鼠肾纤维化模型,采用尾静脉注射的方法整体转染CTGF-siRNA质粒抑制肾脏结缔组织生长因子(CTGF)的表达,观察敲低CTGF表达对肾脏间质纤维化的影响。方法制备UUO模型及活体CTGF基因转染;应用免疫组织化学染色法检测α-平滑肌肌动蛋白(α-SMA)的表达;Western blot检测CTGF及α-SMA蛋白的表达;半定量RT-PCR检测α-SMA mRNA的表达。结果转染CTGF-siRNA质粒的UUO小鼠CTGF及α-SMA的基因表达明显下降(P<0.05)。结论经小鼠尾静脉注射CTGF siRNA质粒能显著抑制小鼠肾CTGF和α-SMA基因表达并能有效抑制小鼠肾脏间质纤维化,提示CTGF siRNA有潜力成为防治肾脏间质纤维化的一种新策略。     

干预CTGF在防治高血压早期肾损害肾脏纤维化中的价值

近年发现,非血流动力学机制在高血压早期肾损害的发病过程中同样起着重要的作用.在非血流动力学诸多因素中,纤维化因子在高血压早期肾损害的发病过程中有着重要意义,其可导致肾脏纤维化(肾小球的硬化和肾小管的间质纤维化).随着研究的深入,AngⅡ、TGFβ1、CTGF、血小板源生长因子、TNFα、ET-1、肝细胞生长因子、干扰素...     

水飞蓟素对肾小管间质纤维化大鼠CTGF的影响

目的探讨水飞蓟素对肾间质纤维化大鼠结缔组织生长因子（CTGF）表达的影响。方法56只雄性SD大鼠,随机分为假手术组,梗阻组,水飞蓟素治疗组。以单侧输尿管结扎术建立输尿管梗阻的动物模型。术后第7、14、21天取术侧肾组织观察病理变化并应用免疫组织化学方法检测肾组织CTGF的表达。结果水飞蓟素可以显著改善梗阻侧肾组织病理变化,下调CTGF表达。结论水飞蓟素可能是通过下调CTGF表达而抑制纤维化的发生,从而起到对肾脏的保护作用。     

结缔组织生长因子在糖尿病肾病发生发展中的作用

目的 动态观察大鼠糖尿病肾病(DN)的不同阶段肾皮质结缔组织生长因子(CTGF)表达的变化及其与肾功能及肾组织形态学变化的相关性,探讨尿液中CTGF含量的检测作为DN诊断指标的意义.方法 以链脲佐菌素(STZ)诱导糖尿病大鼠为研究对象.63只大鼠分为糖尿病组33只和对照组30只.应用逆转录聚合酶链反应、免疫组织化学及蛋白质印迹法研究在糖尿病的不同时期肾皮质CTGF表达的动态变化,并研究其与DN早期肾肥大、ECM的聚积及后期肾小球硬化及肾间质纤维化之间的关系,同时以ELISA法检测糖尿病大鼠尿液中CTGF含量.结果 糖尿病大鼠肾皮质mRNA及CTGF蛋白质表达2周时开始增加,12周时达高峰,分别为(0.905±0.028)、(0.802±0.028),为正常对照组的4.38倍和9.97倍(均P＜0.05);其表达早期出现在肾小球,而后随着病程的发展出现于肾小管间质区域.相关性分析显示CTGF的表达与肾指数、肾小球体积、系膜基质指数、肾小管间质纤维化程度呈正相关(均P＜0.01);糖尿病大鼠尿液中CTGF的含量随着病程的发展逐渐升高,最高为(277.679±10.372)ng/L,与24h尿微量白蛋白、BUN、Cr呈正相关(均P＜0.01).结论 CTGF表达的增加贯穿了DN发病的整个过程,早期介导了肾肥大,肾小球ECM的积聚,晚期又介导了肾小球硬化及肾小管间质纤维化的发生;尿液CTGF的检测可以作为DN诊断的一个指标.     

Inhibition of connective tissue growth factor attenuates paraquat‐induced lung fibrosis in a human MRC‐5 cell line

Chronic exposure to Paraquat (PQ) may result in progressive pulmonary fibrosis and subsequent chronic obstructive pulmonary malfunction. Connective tissue growth factor (CTGF) has been proposed as a key determinant in the development of lung fibrosis. We investigated thus whether knock down of CTGF can prevent human lung fibroblasts (MRC‐5) activation and proliferation with the subsequent inhibition of PQ‐induced fibrosis. MRC‐5 was transfected with CTGF‐siRNAs and exposed to different concentrations of PQ. The siRNA‐silencing efficacy was evaluated using western blotting analyses, qRT‐PCR and flow cytometry. Next, the viability and migration of MRC‐5 was determined. MMP‐2, MMP‐9, and TIMP‐1 accumulation were quantified to evaluate the lung fibrosis exposure to PQ. Over expression of CTGF mRNA was observed in human MRC‐5 cell as early as 6 h following PQ stimulation. CTGF gene expression in MRC‐5 cells was substantially reduced by RNAi, which significantly suppressed the expression of the lung fibrosis markers such as tissue inhibitor of metalloproteinase‐2 (TIMP‐2), Matrix metalloproteinase‐2 (MMP‐2) and Matrix metalloproteinase‐9 (MMP‐9) that were stimulated by PQ. Inhibition of CTGF expression suppressed impeded the proliferation and migration ability of MRC‐5 cells and resulted in cell‐extracellular matrix (ECM) protein accumulation in cells. Our results suggest that CTGF promoted the development of PQ‐induced lung fibrosis in collaboration with transforming growth factor β1 (TGFβ1). Furthermore, the observed arresting effects of CTGF knock down during this process suggested that CTGF is the potential target site for preventing PQ‐induced pulmonary fibrosis. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 1620–1626, 2016.     

CTGF在单侧输尿管梗阻大鼠肾组织中的表达以及活性维生素D_3干预治疗的研究

目的：观察单侧输尿管梗阻大鼠肾脏组织中CTGF的表达以及活性维生素D3（1,25-（OH）2D3）对其表达的影响。探讨1,25-（OH）2D3对大鼠肾间质纤维化的调节作用。方法：随机采用体重180~220g的清洁级Wistar大鼠,雄性,36只,随机分为假手术组（n=12）、UUO模型组（n=12）、活性维生素D3组（n=12）。造膜后活性维生素D3组给予罗盖全（骨化三醇）灌胃（3ng/100g）,假手术组及UUO模型组给予等量生理盐水灌胃。给药后第14天,28天分批（n=6）处死,取造膜侧肾脏。用HE染色及Masson染色方法观察肾脏组织纤维化程度,用免疫组化法测定结缔组织生长因子（CTGF）在各组大鼠造膜侧肾脏的表达情况。结果：与假手术组比较,UUO模型组大鼠的肾脏组织中CTGF的表达有显著增高（P〈0.05）,而在活性维生素D3治疗组中,CTGF的表达低于UUO模型组（P〈0.05）,肾间质纤维化程度也明显小于模型组（P〈0.01）。结论：CTGF在UUO大鼠肾脏中表达明显增高,1,25（OH）2D3能有效抑制UUO大鼠肾脏组织CTGF的表达,从而减轻UUO大鼠肾脏组织纤维化程度。     

罗格列酮对高血压大鼠肾保护作用的研究

目的：观察结缔组织生长因子（CTGF）在两肾-夹高血压大鼠肾间质中的表达及其与肾损害的关系,探讨罗格列酮对CTGF在肾间质表达的影响及高血压肾保护作用的机制。方法：两肾-夹高血压大鼠随机分为阳性对照组（B组）、罗格列酮治疗组（C组）,假手术组作为阴性对照组（A组）。监测各组大鼠各阶段血压的变化,检测尿＆微球蛋白（-MG）、尿素氮（Bun）、血肌酐（Cr）。免疫组化法检测CTGF在肾组织中的表达,并判断肾组织的病理学变化及肾间质纤维化程度。结果：治疗前B组与C组的尿B．MG均较A组明显升高（P〈0．01）,B组与C组之间的差异无统计学意义（P〉0．05）,而治疗后C组的尿＆-MG较B组明显降低（P〈0．05）;CTGF在B组和c组的表达较A组均明显增加（P〈0．01）,而C组CTGF的表达较B组降低（P〈0．05）。CTGF的表达与β2-MG及肾间质纤维化呈正相关（r=0．734,P〈0．01;r=0．777,P〈0．01）。结论：罗格列酮可以降低血压,减少蛋白尿,下调CTGF在肾间质的表达,从而延缓肾间质纤维化（RIF）的发展,发挥肾保护作用。     

Dual Therapeutic Effects of an Albumin-Based Nitric Oxide Donor on 2 Experimental Models of Chronic Kidney Disease

Chronic kidney disease (CKD) is accompanied by a variety of complications, typically renal anemia and kidney fibrosis. Accordingly, it is desirable to develop the novel therapeutics that can treat these CKD conditions. Since nitric oxide (NO) has multiple functions including hypoxia inducible factor stabilizing, anti-inflammatory, anti-oxidative, and anti-apoptoic activities, the use of NO for the CKD therapy has attracted considerable interest. Here, we evaluate the therapeutic impacts of S-nitrosated human serum albumin (SNO-HSA), a long-lasting NO donor, on 2 animal models of CKD. SNO-HSA increased the expression of erythropoietin (EPO), VEGF, and eNOS by stabilizing hypoxia inducible factor–1α in HepG2 and HK-2 cells. SNO-HSA increased hematopoiesis in both healthy and renal anemia rats, suggesting the promotion of EPO production. In unilateral ureteral obstruction–treated mice, SNO-HSA ameliorated kidney fibrosis by suppressing the accumulation of renal extracellular matrix. SNO-HSA also inhibited unilateral ureteral obstruction–induced α–smooth muscle actin increase and E-cadherin decrease, suggesting that SNO-HSA might suppress the accumulation of myofibroblasts, an important factor of fibrosis. SNO-HSA also inhibited the elevations of fibrosis factors, such as transforming growth factor–β, interleukin-6, and oxidative stress, while it increased EPO production, an anti-fibrosis factor. In conclusion, SNO-HSA has the potential to function as a dual therapeutics for renal anemia and kidney fibrosis.     

罗格列酮对糖尿病大鼠肾脏结缔组织生长因子表达和排泄的影响

目的探讨罗格列酮对糖尿病大鼠肾脏结缔组织生长因子(CTGF)表达和排泄的影响。方法Wistar大鼠分为正常对照组、糖尿病模型组、糖尿病罗格列酮(4mg.kg-1.d-1)治疗组,每组8只,检测各组第1、4、6、8周尿CTGF排泄率(ELISA法),进行组间差异比较;于第8周处死大鼠,每组各5只,应用免疫组化检测肾皮质CTGF蛋白表达。结果第4、6、8周糖尿病大鼠治疗组及糖尿病模型组尿CTGF排泄率及第8周肾皮质CTGF蛋白表达水平较正常组明显升高,治疗组上述指标均较模型组低(P<0.01)。结论罗格列酮可抑制肾脏CTGF表达和排泄,这可能与其保护肾脏有关。     

灯盏花素对大鼠糖尿病模型肾组织TGF-β1与CTGF表达影响的实验研究

目的　探讨灯盏花素对大鼠糖尿病模型肾组织转化生长因子 β1(TGF β1)与结缔组织生长因子 (CTGF)表达的影响。方法　建立STZ诱导的单侧肾切除糖尿病模型 ,随机分 :对照组、模型组、灯盏花素给药组 (2 0mg·kg-1·d-1,灌胃 )。观察 8wk后大鼠体重、肾重、肾重 /体重、2 4h尿白蛋白排泄率 (AER)、肾小球面积 (AG)和容量 (VG)及系膜区面积 (AM)的变化 ,并检测肾组织与尿MDA含量及肾组织SOD、CAT、GSH PX活性 ,通过免疫组化检测肾小球TGF β1与CTGF蛋白表达。结果　灯盏花素给药对大鼠糖尿病模型血糖、体重无明显影响 ,可明显抑制肾重、肾重 /体重、AER、AG、VG 及AM 的增加 (P <0 0 5 ) ;对肾组织及尿MDA含量的增加有明显抑制作用 (P <0 0 5 ) ,并明显提高肾组织SOD、CAT及GSH PX活性 (P <0 0 5 ,P <0 0 1)。免疫组化半定量分析显示 ,与对照组相比糖尿病大鼠肾小球TGF β1及CTGF表达明显增加 (P <0 0 1) ,灯盏花素给药对其有明显抑制作用 (P <0 0 5 )。结论　灯盏花素对大鼠糖尿病模型肾脏有明显保护作用 ,其机制部分与抑制肾组织TGF β1与CTGF过高表达有关     

肝细胞生长因子对心肌成纤维细胞结缔组织生长因子表达的影响

目的通过体外培养的新生大鼠心肌成纤维细胞研究肝细胞生长因子(HGF)对结缔组织生长因子表达(CTGF)的影响。方法差速贴壁法分离新生SD大鼠心肌成纤维细胞(CFs)及免疫组化法对其进行鉴定,选AngⅡ(10~(-6)mol/L)作诱导浓度,与HGF100ng/ml孵育48h,采用反转录聚合酶链反应(RT-PCR)技术测定CFsCTGFmRNA的表达,WesternBlot法测定CTGF蛋白的表达。结果肝细胞生长因子能够在mRNA及蛋白水平上大部分抑制AngⅡ对CTGF表达的诱导。结论初步证实在心肌纤维化的过程中,HGF与CTGF是一对拮抗生长因子,HGF可能通过抑制CTGF表达的途径逆转心肌纤维化。     

血清结缔组织生长因子在肝纤维化评估中的临床价值

目的探讨血清结缔组织生长因子(CTGF)和肝纤维化的关系,评估结缔组织生长因子在肝纤维化诊断中的临床价值。方法应用酶联免疫吸附法(ELISA)测定血清CTGF水平,探讨血清CTGF浓度和肝纤维化程度的关系;通过CTGF和一些已经证实的肝纤维化标记物的ROC曲线分析,评估CTGF诊断肝纤维化的价值。结果血清CTGF水平和肝纤维化分期明显相关,r为0.689(P<0.001),CTGF用于鉴别轻微纤维化和明显纤维化的ROC曲线下面积为0.841,95%可信区间0.762~0.920。结论血清CTGF和肝纤维化分期明显相关,提示血清CTGF水平可能是预测肝纤维化分期比较好的标记物,血清CTGF在肝纤维化的评估中是一个非常有价值的指标。     

Connective tissue growth factor: A role in airway remodelling in asthma?

1. Severe persistent asthma is accompanied by structural changes in the airway, referred to as remodelling. The mechanisms driving airway remodelling are poorly understood. 2. Transforming growth factor (TGF)-beta is increased in the airways of patients with asthma. Many of the effects of TGF-beta are mediated by connective tissue growth factor (CTGF). 3. Overexpression of CTGF is linked to many fibrotic diseases, but its exact role in airway remodelling is unknown. 4. Connective tissue growth factor mediates cell adhesion, migration, proliferation, survival, extracellular matrix synthesis and has a role in angiogenesis. 5. Current asthma therapies do not inhibit CTGF induction. 6. Understanding the mechanisms underlying the role of CTGF in airway remodelling may lead to new therapeutic strategies for asthma.     

醛固酮对人近端肾小管上皮细胞结缔组织生长因子mRNA表达的影响

目的探讨醛固酮(ALD)在体外是否能够促进人近端肾小管上皮细胞株(HK2)结缔组织生长因子(CTGF)的表达.方法采用体外培养的HK2细胞,分别观察不同浓度(10-11、10-10、10-9、10-8、10-7 mol/L)ALD处理48 h,或用10-9 mol/L ALD处理不同时间(24 h、48 h、72 h)对HK2细胞CTGF表达的影响,应用RT-PCR法检测CTGFmRNA水平的表达.结果HK2细胞培养48 h后,空白对照组即有CTGF的基础表达,ALD不同浓度组CTGF mRNA的表达均有增加,与空白对照组相比较差异有统计学意义(P＜0.05或P＜0.01);以10-9 mol/L浓度的ALD作用于HK2细胞,随着作用时间的延长,CTGF mRNA的表达量明显增加,与对照组相比较差异有统计学意义(P＜0.05或P＜0.01).结论ALD能够促进HK2细胞CTGF mRNA水平的表达,并具有剂量依赖性和时间依赖性,ALD可能通过促进CTGF的分泌,发挥其促进肾间质纤维化的作用.     

结缔组织生长因子在病理性瘢痕成纤维细胞中的表达

要目的探讨结缔组织生长因子(CTGF)人类增生性瘢痕成纤维细胞中的表达与瘢痕纤维化之摘在间的关系。方法应用半定量逆转录聚合酶链反应(RT-PCR)术,测了瘢痕组织原代培养的成纤维细技检胞中CTGFmRNA表达。结果CTGFmRNA在人类增生性瘢痕成纤维细胞中高度表达,与正常皮肤组织的成纤维细胞比较,有显著性差异(P<0.01)。结论CTGF在人类增生性瘢痕成纤维细胞中高度表达,提示其在增生性瘢痕的纤维化进程中起着重要的作用,这为临床上寻找安全、有效的治疗增生性瘢痕提供了新的思路。     

辛伐他汀抑制大鼠炎症性肠病结肠纤维化结缔组织生长因子的表达

目的探讨大鼠炎症性肠病(IBD)结肠纤维化结缔组织生长因子(CTGF)的表达及辛伐他汀对其影响。方法 32只SD大鼠均分为对照组、IBD模型对照组、辛伐他汀低剂量(5mg/kg)和高剂量(20mg/kg)干预组。21d后观察结肠病理组织学变化;RT-PCR和Western blot检测结肠Ⅰ型胶原、CTGF mRNA和蛋白表达。结果与对照组相比,模型组结肠黏膜及黏膜下层见大量炎症细胞浸润,纤维组织增生,Ⅰ型胶原和CTGF表达增多(P<0.01)。辛伐他汀干预后,病理组织学较模型组改善,Ⅰ型胶原和CTGF表达明显下降(P<0.05)。结论 CTGF参与结肠炎大鼠肠纤维化形成,辛伐他汀可能通过抑制CTGF的表达减轻IBD肠纤维化进展。     

Rosiglitazone inhibits angiotensin II-induced CTGF expression in vascular smooth muscle cells - role of PPAR-gamma in vascular fibrosis

Angiotensin (Ang) II plays a pivotal role in vascular fibrosis, which leads to serious complications in hypertension and diabetes. Connective tissue growth factor (CTGF) is a potent profibrotic factor implicated in the Ang II-induced pathologic fibrosis process. PPAR-gamma activators thiazolidinediones have been recently reported to have beneficial vascular effects. However, their effects and related molecular mechanisms on extracellular matrix (ECM) turnover in vascular smooth muscle cells (VSMCs) are unknown. The present study evaluated the regulation of Ang II-induced CTGF, ECM production and cell growth by rosiglitazone in VSMCs. In aorta of Ang II-infused rats, CTGF expression was markedly increased, and type III collagen and fibronectin overexpression was observed. Cotreatment with rosiglitazone diminished these changes, whereas increased nuclear PPAR-gamma expression in VSMCs. In growth-arrested VSMCs, rosiglitazone attenuated the proliferation and apoptosis, increased PPAR-gamma production and activation, and reduced CTGF and ECM production in response to Ang II in a dose-dependent fashion. These inhibitory effects were attenuated by the pretreatment of cells with PPAR-gamma antagonist GW9662 or bisphenol A diglycidyl ether (BADGE). Furthermore, rosiglitazone inhibited Ang II-induced Smad2 production and phosphorylation but had no effect on transforming growth factor-beta(1) (TGF-beta(1)) expression. These results suggest that in Ang II-stimulated VSMCs, rosiglitazone caused an antiproliferative, antiapototic effect and reduces ECM production through mechanisms that include reducing CTGF expression, and a crosstalk between PPAR-gamma and Smad may be involved in the inhibitory effects of rosiglitazone. This novel finding suggests a role of PPAR-gamma activators in preventing Ang II-induced vascular fibrosis.     

牛蒡子总木脂素对自发性糖尿病大鼠肾脏病变的影响

目的观察牛蒡子总木脂素对自发性糖尿病大鼠(GK大鼠)肾脏病变的影响。方法 20只GK大鼠造模成功后随机分为模型组和牛蒡子总木脂素组(300 mg·kg~(-1),ig,2次·d-1),同时采用10只同龄Wistar大鼠作为正常组。给药期间观察大鼠生长状况,每周监测体重。给药12 wk末,测定肾脏肥大指数、血尿素氮(BUN)、血肌酐(SCr)、空腹血糖(FBG)、糖化血红蛋白(HbA1c)、尿蛋白和尿微量白蛋白。并观察各组肾脏病变状况,采用Western blot法检测肾皮质中转化生长因子β_1(TGF-β1)和结缔组织生长因子(CTGF)的表达水平。结果至实验结束,3组动物的体重增长无显著差异。总木脂素组肾脏肥大指数为(3.62±0.10)×10~(-3),低于模型组[(4.50±0.49)×10~(-3),P<0.05],与正常组无显著差异[(3.27±0.31)×10~(-3),P>0.05],光镜下总木脂素组肾脏病理改变较模型组轻微。总木脂素组大鼠SCr、BUN、HbA_(1c)、FBG及尿微量白蛋白水平均低于模型组(P<0.01),其中SCr、BUN、HbA_(1c)及尿微量白蛋白水平与正常组无显著差异(P>0.05)。总木脂素组肾皮质中TGF-β_1和CTGF蛋白表达水平(22.98±5.08和21.30±2.21)低于模型组(26.58±2.35和24.93±4.26,P<0.05),TGF-β_1水平接近正常组(22.32±2.49,P>0.05),CTGF仍显著高于正常组(17.18±3.64,P<0.01)。结论牛蒡子总木脂素可以减轻GK大鼠肾脏病理改变,改善肾脏功能,可能与抑制肾脏中TGF-β_1和CTGF的表达有关。