单侧输尿管梗阻大鼠肾脏转化生长因子β受体亚型表达的变化

目的 探讨转化生长因子β（TGF-β）Ⅰ型受体（RⅠ）、Ⅱ型（RⅡ）受体以及下游Smad蛋白在单侧输尿管梗阻（UUO）大鼠模型肾脏中表达及意义。 方法 90只雌性Wistar大鼠随机分为正常对照组（CON组）、假手术组（SOR组）和单侧输尿管梗阻组（UUO组）,分别于术后1、3、7、14、21 d处死,检测各组大鼠肾功能;PAS与Masson染色观察大鼠肾间质病理形态改变;实时定量PCR基因芯片分析正常大鼠和肾间质纤维化大鼠肾组织TGF-βⅠ、Ⅱ、Ⅲ型受体及Smad蛋白家族表达。筛选出差异表达的受体亚型,进一步应用实时荧光定量PCR、蛋白免疫印迹法、免疫荧光法检测和验证筛选出的差异受体亚型在不同分期肾间质纤维化大鼠肾组织的分布和表达。 结果 与CON组相比,UUO组大鼠的Scr及BUN于术后3 d开始升高（P < 0.05）,第21天达峰值（P < 0.01）;UUO组术后3 d肾间质可见明显炎性细胞浸润;14 d后出现明显肾小管萎缩;21 d可见明显肾间质纤维化。UUO组肾组织TGF-βⅠ型受体ALK-5、ALK-7和TGF-βRⅡ的mRNA表达于术后3 d上升并随梗阻时间延长逐渐增加（P < 0.05）,于14 d达到峰值（均P < 0.01）;ALK-6的mRNA表达于术后3 d下降（P < 0.05）并随梗阻时间延长逐渐减少,于14 d达谷值（P < 0.01）。ALK-5、ALK-6、ALK-7和TGF-βRⅡ蛋白表达与基因表达一致。Smad2/3及磷酸化（p）-Smad2/3的蛋白表达于术后3 d上升（均P < 0.05）并随梗阻时间延长逐渐增加,于14 d达到峰值（均P < 0.01）。 结论 在肾间质纤维化进展中不同TGF-β受体亚型存在不同的变化规律并与肾间质纤维化进展密切关联。     

肝细胞生长因子在单侧输尿管梗阻大鼠肾脏中的表达及其意义

目的　检测肝细胞生长因子 (HGF)在单侧输尿管梗阻 (UUO)大鼠肾脏中的表达并探讨其与小管细胞增殖与凋亡的关系。方法　免疫组织化学和脱氧核苷酸末端转移酶介导的核苷酸缺口末端标记 (TUNEL)法分别检测了第 0、3、6、9、1 2天UUO大鼠肾脏小管间质α 平滑肌肌动蛋白 (α SMA)、转化生长因子 β1 (TGF β1 )、HGF的表达以及小管细胞增殖性细胞核抗原 (PCNA)的表达和细胞凋亡的情况。噻唑蓝 (MTT)法检测重组HGF(rHGF)对培养的小管上皮细胞促增殖作用。结果　随着梗阻时间的延长 ,UUO大鼠肾脏α SMA、TGF β1蛋白的表达逐渐增多 ,HGF与PCNA蛋白的表达在第 6天达到高峰 ,随后逐渐下降 ,且它们之间呈正相关 (r =0 .870 5 ,P <0 .0 1 ) ,而细胞凋亡数随着时间的延长也逐渐增多。rHGF可促进体外培养的小管上皮细胞增殖。结论　HGF具有促进小管上皮细胞增殖的作用 ,同时它可能具有抗凋亡的作用     

血管内皮生长因子在肾脏疾病中的研究进展

血管内皮生长因子（VEGF）是一种促进血管内皮生长及渗透的分子，在血管渗透、血管的生长过程中起重要作用。检测VEGF的表达，对探讨肾脏疾病的发病机制、观察病情进展、指导临床治疗及判断预后有重要意义。     

大鼠缺血后肢骨骼肌血管内皮生长因子及其受体的表达

目的 研究大鼠缺血后肢侧枝代偿和血管内皮生长因子（VEGF）及其受体表达的动态变化。为外源性VEGF治疗下肢缺血性疾病提供理论依据。方法 切除SD大白鼠右后肢全长股动脉，随机分为9个时间组：造模后1、3d、1、2、3、4、6、8及12周，各组5只动物。分别于造模前后和观察期末检测双后肢大、小腿肌肉Fit-1、Flk-1蛋白及mRNA表达，各组观察期末实验动物后肢动脉DSA检查。结果 （1）缺血后3d，5只大鼠右后肢出现溃疡（11．11％）；2周后，4只大鼠后肢溃疡愈合，而1只趾端坏疽（2．22％）。（2）缺血后2周，患肢侧枝形成达到高峰，12周时仍可见侧支血管显影。（3）缺血早期（3周内），VEGF及其受体的表达均较健侧显著增强（P〈0．05）；缺血中期（3～8周）。VEGF和Flt-1表达迅速下降，Flk-1仍表达；缺血后期（8周后），VEGF及其受体的表达均低至极低水平，与对侧差异无统计学意义（P〉0．05）。结论（1）肢体缺血后自身的血管新生不能完全满足缺血组织的需要。（2）缺血早期外源性的VEGF补充是不必要的；缺血中期补充VEGF是适宜的；缺血后期在应用VEGF治疗的同时，也需要干预受体的表达。     

槲皮素对单侧输尿管梗阻大鼠肾脏α-平滑肌肌动蛋白表达的影响

目的:观察槲皮素(Que)对单侧输尿管梗阻大鼠肾脏病理和α-平滑肌肌动蛋白(α-SMA)表达的影响,探讨槲皮素对肾间质纤维化的保护作用及作用机制.方法:采用单侧输尿管结扎(UUO)致肾间质纤维化大鼠模型.将36只大鼠随机分为3组(n=12):假手术组(Sham组)、模型组(UUO组)、槲皮素(剂量100 mg·kg-1·d-1)治疗组(Que组).术后10 d以化学比色法测定梗阻侧肾组织匀浆中羟脯氨酸(Hyp)的含量;HE和Masson染色观察大鼠肾脏病理变化;采用免疫组织化学法、Western blot法检测各组大鼠梗阻侧肾脏α-SMA的表达部位及蛋白表达水平.结果:HE和Masson染色联合图像分析显示大鼠UUO术后10 d梗阻侧肾脏呈现肾间质纤维化,经 Que治疗梗阻侧肾脏病理改变减轻(P＜0.05).免疫组织化学和免疫蛋白印记分析显示经Que治疗梗阻侧肾脏α-SMA染色阳性表达范围和相对表达丰度明显减少(P＜0.01).同时经Que治疗后肾组织匀浆中的Hyp含量降低(P＜0.05).结论:槲皮素可以减轻UUO大鼠肾间质纤维化,其作用机制与抑制肾小管上皮细胞转化成肌成纤维细胞有关.     

大豆异黄酮对单侧输尿管梗阻大鼠肾间质纤维化的影响

大豆异黄酮是近年来国内外研究较多的一种具有抗细胞增殖、抗癌与抗衰老作用的非营养成分,现有资料表明,异黄酮还有抗炎、抗氧化的作用犤1犦,可以抑制单核/巨噬细胞的活化和迁移以及抑制细胞间黏附分子(ICAM-1)的表达犤2犦。本实验以大鼠单侧输尿管梗阻模型(UUO)为研究对象,研究异黄酮对梗阻肾病理改变、ICAM-1表达及巨噬细胞浸润的影响,初步探讨异黄酮对肾间质纤维化(RIF)的抑制作用及其机制。一、材料与方法1.动物模型:雄性Wistar大鼠,6周龄,体重180～200g,随机分为假手术组(S)18只、对照组(C)30只和异黄酮组(G)42只。大鼠UUO模…     

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。     

缬沙坦对单侧输尿管梗阻大鼠肾脏氧化应激的影响

目的：探讨血管紧张素Ⅱ-1型受体拮抗剂（AT1Ra）缬沙坦对单侧输尿管梗阻（UUO）大鼠肾脏氧化应激的影响。方法：Wistar大鼠行左侧输尿管结扎术,分为UUO模型组（n=11）,缬沙坦治疗组（n=11）,同时设假手术对照组（n=7）。术后第14天处死各组大鼠,进行HE和Masson染色,观察肾脏病理变化;比色法测定肾组织丙二醛（MDA）和超氧化物歧化酶（SOD）含量;免疫组织化学方法测定α-平滑肌肌动蛋白（α-SMA）和转化生长因子-β1（TGF-β1）的表达。结果：UUO组与假手术组大鼠比较,肾脏病理改变加重,肾组织MDA含量明显升高（P〈0.01）,SOD含量下降（P〈0.05）,肾组织α-SMA和TGF-β1的表达显著增加（P〈0.01）。缬沙坦治疗组与UUO组大鼠比较,肾间质纤维化减轻（P〈0.05）,肾组织MDA含量降低（P〈0.05）,SOD含量升高（P〈0.05）;同时肾组织α-SMA和TGF-β1的表达降低（P〈0.05）。结论：缬沙坦可通过减少UUO组肾组织脂质过氧化物的产生,增加抗氧化酶的含量,下调TGF-β1的表达,从而显著改善UUO所致的肾间质纤维化。     

血管内皮生长因子及其受体在肾脏疾病中的研究进展

近来有较多报道，血管内皮生长因子（vascular endothelial growth factor，VEGF）是一种促进内皮细胞的增殖、迁移，增强血管通透性的细胞因子，参与微血管病变的病理过程。VEGF在肾脏组织中含量丰富，主要由肾小球足细胞合成分泌。VEGF通过与表达于内皮细胞及足细胞上的VEGF受体结合以旁分泌与自分泌的方式而发挥重要的生理作用，包括。肾脏的胚胎发育、维持肾小球滤过屏障功能与结构的完整性、肾脏血管的再生与修复。局部的肾组织VEGF及其受体的表达变化参与了多种肾脏疾病的发生、发展。现就有关VEGF及其受体与。肾脏疾病的研究作简要综述。     

福辛普利对单侧输尿管梗阻大鼠肾脏保护作用机制研究

肝细胞生长因子(HGF)是一种具有多种生物学效应的蛋白。研究表明，重组HGF或HGF基因治疗可以明显延缓肾小管间质纤维化的进展。我们利用单侧输尿管使阻(UUO)模型观察ACEI福辛普利对大鼠输尿管使阻后肾脏HGF、转化生长因子β1(TGF-β1)及增殖细胞核抗原(PCNA)表达的影响，从而为ACEI在减轻肾问质炎症和纤维化中的应用提供新的理论依据。     

坎地沙坦对单侧输尿管梗阻大鼠肾间质纤维化及肾脏骨桥蛋白表达的影响

目的 观察坎地沙坦(CAN)对单侧输尿管梗阻(UUO)大鼠肾间质纤维化的影响,并观察骨桥蛋白(OPN)与肾间质纤维化的关系及CAN干预对肾脏OPN表达的影响,探讨其在纤维化中的作用机制.方法 (1)36只成年雄性SD大鼠随机分为3组,每组12只:假手术组(Sham)、UUO模型组(UUO)、坎地沙坦治疗组(CAN).UUO模型组和CAN组大鼠行左侧输尿管结扎术,Sham组只游离左侧输尿管但不结扎.CAN组于术前1 d开始用CAN治疗[10 mg/(kg·d)]灌胃.术后第7、14天分别处死大鼠,左侧肾脏组织行Masson染色,免疫组织化学方法检测肾组织中OPN的表达,逆转录-聚合酶链反应(RT-PCR)检测OPN mRNA表达水平.结果 Masson染色结果显示术后7 d和14 d UUO组大鼠肾纤维化阳性面积分别为15.2%和24.8%,CAN组为1O.1%和18.5%.CAN组与UUO组大鼠术后7 d和14 d大鼠肾纤维化阳性面积差异有统计学意义(P＜0.05).UUO模型大鼠OPN蛋白及mRNA水平均较Sham组明显升高(P＜0.01),CAN组OPN蛋白及mRNA水平较UUO组明显降低(P＜0.05),但较Sham组高(P＜0.01).结论 CAN能有效地延缓UUO大鼠肾间质纤维化的进展,其延缓肾间质纤维化作用可能与下调OPN蛋白和mRNA有关.     

Therapeutic implications of mesenchymal stem cells transfected with hepatocyte growth factor transplanted in rat kidney with unilateral ureteral obstruction

Purpose

The purpose of the study was to establish whether bone marrow mesenchymal stem cells (MSCs) transfected with hepatocyte growth factor (HGF) can migrate and localize in the rat's kidney with unilateral ureteral obstruction (UUO) and contribute to repair of renal fibrosis.

Methods

We separated and cultured bone marrow-derived MSCs of male rats in vitro and transfected them with adenovirus-mediated HGF (Ad-HGF). The expression of HGF was measured with enzyme-linked immunosorbent assay. Sixty female rats were sham operated (n = 24) or subjected to left UUO: Ad-HGF-transfected MSCs, uninfected MSCs, or saline was injected into the rat's tail vein. Kidney tissue was collected at the end of the seventh or 14th day after operation. The distribution of Y chromosome in the kidney after Ad-HGF-transfected MSCs transplantation was determined by an in situ hybridization method. As the hallmark of myofibroblasts, α-smooth muscle actin (expression of which significantly increases in the presence of renal fibrosis) was detected by immunohistochemistry in all UUO rats' left kidney tissue.

Results

Y chromosome-positive cells were found only in the obstructed kidney of the transplantation group. The positive cells were mainly distributed in the tubular cells. The average intensity of immunolabeling for α-smooth muscle actin in the transplanted group significantly decreased compared with sham-transplanted group (P < .05), and the expression in the rats injected with uninfected MSCs was higher than that in the rats with MSCs transfected with HGF (P < .05).

Conclusions

Mesenchymal stem cells transfected with HGF can migrate to the rat kidney with UUO and are mainly distributed in the region of renal tubular epithelial cells. The data indicate that MSCs transfected with HGF contribute to a reduction of renal fibrosis after ureteral obstruction and suggest that this may be exploited therapeutically.     

The role of autophagy in unilateral ureteral obstruction rat model

Aim: Autophagy is a cellular process of degradation of damaged cytoplasmic components and regulates cell death or proliferation. Unilateral ureteral obstruction (UUO) is a model of progressive renal fibrosis in the obstructed kidney. And UUO is followed by compensatory cellular proliferation in the contralateral kidney. We investigate the role of autophagy in the obstructed kidney and contralateral kidney after UUO. Methods: To obtain the evidence and the patterns of autophagy during UUO, the rats were sacrificed 3, 7 and 14 days after UUO. To examine the efficacy of the autophagy inhibitors, 3‐methyladenine (3‐MA), the rats were treated daily with intraperitoneal injection of 3‐MA (30 mg/kg per day) for 7 days. Results: After UUO, autophagy was induced in the obstructed kidney in a time‐dependent manner. Inhibition of autophagy by 3‐MA enhanced tubular cell apoptosis and tubulointerstitial fibrosis in the obstructed kidney after UUO. In the contralateral kidney, autophagy was also induced and prolonged during UUO. Inhibition of autophagy by 3‐MA increased the protein expression of proliferating cell nuclear antigen significantly in the contralateral kidney after UUO. The Akt‐mammalian target of rapamycin (mTOR) signalling pathway was involved in the induction of autophagy after UUO in both kidneys. Conclusion: Our present results support that autophagy induced by UUO has a renoprotective role in the obstructed kidney and regulatory role of compensatory cellular proliferation in the contralateral kidney through Akt‐mTOR signalling pathway.     

Expression of renin and its mRNA in the adult rat kidney with chronic ureteral obstruction

Angiotensin II has been implicated in mediating renal vasoconstriction resulting from chronic unilateral ureteral obstruction (UUO) in both mature and developing animals. We have previously shown that chronic neonatal UUO results in increased distribution of renin and its mRNA in the obstructed kidney, as well as of immunoreactive renin in the intact opposite kidney. The present study was designed to evaluate the effects of 24 hours versus 4 weeks of UUO on the distribution of renin mRNA and its protein in the adult rat kidney. Renin was detected by immunocytochemistry using a polyclonal anti-rat renin antibody. Renin mRNA was localized by in situ hybridization to an oligonucleotide complementary to renin mRNA. UUO of 24 hours' or 4 weeks' duration did not alter the distribution of renin and its mRNA in the obstructed kidneys as compared with sham-operated kidneys, although kidneys obstructed for 4 weeks had a significant increase in the percent of renin-containing juxtaglomerular apparatuses (JCA) when compared with the intact opposite kidneys (P less than 0.05). Compensatory hypertrophy was not present in the intact opposite kidneys after 24 hours of UUO and distribution of renin gene expression was not altered at that time. However, 4 weeks following contralateral UUO, the intact kidneys were hypertrophied and showed a decrease in renin gene expression relative to the obstructed and sham-operated kidneys. We conclude that unlike UUO during early development, chronic UUO in the mature animal does not activate renin gene expression nor alter renin distribution in the obstructed kidneys. Renin gene expression is suppressed in the hypertrophied kidney with prolonged contralateral UUO.(ABSTRACT TRUNCATED AT 250 WORDS)     

罗格列酮对单侧输尿管梗阻大鼠肾脏的保护作用

目的观察罗格列酮对单侧输尿管梗阻(UUO)大鼠肾皮质过氧化物酶体增殖物活化受体(PPAR)γ、转化生长因子(TGF)β1表达介导的肾间质纤维化的作用。方法UUO大鼠给予罗格列酮5mg·kg-1·d-1灌胃,用免疫组化、RT-PCR及Western印迹的方法检测术后7d、14dPPARγ、TGF-β1、增殖细胞核抗原(PCNA)表达量及观察肾脏病理改变。结果与假手术组相比,UUO组及药物治疗组PPARγ、TGF-β1、、PCNA表达均增高且UUO组显著高于治疗组(P<0.05)。结论罗格列酮可通过活化PPARγ,下调TGF-β1,从而减轻UUO术后肾组织间质纤维化。     

大黄酸对单侧输尿管梗阻大鼠肾脏的抗氧化保护作用

目的观察大黄酸（RH）对单侧输尿管梗阻（UUO）大鼠肾间质损伤的抗氧化保护作用。方法将30只雄性SD大鼠分成假手术组（Sham组）6只;UUO模型组（UUO组）和RH干预组（UUO＋RH组）各12只。除Sham组外,UUO组和UUO＋RH组分别在第3、7天测定大鼠左肾皮质匀浆中脂质过氧化物标志物丙二醛（MDA）及抗氧化酶过氧化氢酶（CAT）和超氧化物歧化酶（SOD）含量。结果UUO组较Sham组肾脏病理改变加重、肾组织MDA含量升高（P〈0.05）、SOD和CAT含量下降（P〈0.05）;UUO＋RH组较UUO组肾间质纤维化程度减轻、。肾组织MDA含量降低（P〈0.01）、SOD和CAT含量升高（P〈0.01）。结论RH能减少单侧输尿管梗阻侧肾皮质脂质过氧化物的产生,同时增加抗氧化酶的含量,通过改善UUO大鼠肾脏氧化应激来发挥肾脏保护作用。     

螺内酯对单侧输尿管梗阻大鼠肾脏的保护作用

目的观察螺内酯对单侧输尿管梗阻(UUO)大鼠肾皮质金属蛋白酶1组织抑制剂(TIMP-1)介导的肾间质纤维化的影响。方法UUO大鼠给予螺内酯20mg·kg-1·d-1灌胃,用免疫组化及RT-PCR的方法检测术后7d、14d、21d的TIMP-1表达量并观察肾脏病理改变。结果与假手术组相比,UUO组及螺内酯组的TIMP-1mRNA和TIMP-1蛋白表达水平均明显增高(P<0.01),且UUO组显著高于螺内酯组(P<0.01)。结论螺内酯可通过下调TIMP-1减轻UUO术后肾组织间质纤维化。     

血管内皮生长因子及其受体mRNA在乳腺癌组织中的表达

目的　研究血管内皮生长因子及其受体FLT 1、FLK 1mRNA在乳腺癌组织中的表达及与临床病理因素的相关性。　方法　采用逆转录聚合酶链反应技术对手术的 47例乳腺癌标本 ,11例乳腺良性病变标本中血管内皮生长因子 (VEGF)及其受体FLT 1、FLK 1mRNA的表达进行检测。结果　在良、恶性乳腺组织中均检测到VEGF12 1、165mRNA的表达 ,在乳腺癌组织中表达水平分别为0 42 0± 0 13 3、0 2 91± 0 0 94高于良性乳腺组织 0 196± 0 0 67(P =0 0 0 0 )、0 2 0 6± 0 0 5 8(P =0 0 0 1) ;在乳腺癌组织中 ,VEGF12 1mRNA表达高于VEGF165mRNA(P =0 0 0 0 ) ,而在良性乳腺组织中两者表达水平差异无显著性意义 (P =0 666)。FLT 1、FLK 1mRNA在部分乳腺癌组织中表达 ,分别为 3 8 3 %(18/4 7)和 2 5 5 % (12 /4 7) ,而在良性乳腺组织中未见表达。乳腺癌组织中VEGF12 1、VEGF165、FLT 1、FLK 1mRNA表达与患者年龄、肿块大小、淋巴结转移状况、肿瘤分期及雌、孕激素受体状况之间无明显相关性。　结论　乳腺癌组织中VEGF12 1、165及其受体FLT 1、FLK 1mRNA表达水平上调 ,提示其在乳腺癌的血管生成中起着重要作用。     

Increased E-cadherin expression in the ligated kidney following unilateral ureteric obstruction

E-cadherin expression in the kidney is used as a surrogate marker of epithelial mesenchymal transition for the testing of various antifibrotic strategies. Here we reexamined E-cadherin expression in the kidneys of rats with unilateral ureteric obstruction, which was previously reported to decrease in parallel with the development of tubulointerstitial disease in this widely used experimental model of renal fibrosis and epithelial mesenchymal transition. E-cadherin mRNA expression was consistently increased both acutely (hours) and chronically (days) in the ligated kidney compared to the cognate non-ligated kidney. Increased E-cadherin protein levels were also found in the ligated kidney particularly in dilated tubular segments. Simulation of early pressure changes in the ligated kidney by mechanical stretch of human renal epithelial cells in culture did not alter E-cadherin expression. Porcine LLCPK-1 cells subjected to hypotonic stretch, however, did have increased E-cadherin mRNA and protein levels, responses that were not prevented by transforming growth factor-beta, a cytokine that promotes epithelial mesenchymal transition. Our findings question the utility of E-cadherin as a marker of epithelial mesenchymal transition in this model of renal fibrosis.