苦参素对单侧输尿管梗阻大鼠肾间质纤维化的保护作用

目的探讨苦参素对单侧输尿管梗阻(UUO)大鼠肾间质纤维化的影响及可能机制。方法45只雄性SD大鼠随机分为3组:A假手术组,B单侧输尿管结扎(UUO)组,C治疗组。治疗组在UUO的基础上每天以苦参素100mg/Kg/d腹腔注射。B组和C组于术后第7,14,21,28分别处死5只大鼠,A组于第28天处死大鼠。用PAS及Masson染色法观察肾脏病理改变。用免疫组化法检测转化生长因子β1(transforming growth factor-beta1 TGF-β1),α-平滑肌肌动蛋白(alpha-smooth muscle actin-αSMA),Ⅰ型胶原(collagenⅠColⅠ)的表达。结果与UUO组相比,治疗组梗阻侧肾脏TGF-β1,-αSMA,ColⅠ的表达明显减少,肾小管损害和肾间质纤维化的程度也明显减轻。结论苦参素可通过下调TGF-β1,减少肾小管上皮细胞转分化而减轻肾间质纤维化。     

单侧输尿管梗阻法制作大鼠肾间质纤维化模型的改进

目的建立改良的大鼠肾间质纤维化模型。方法用单侧输尿管结扎术建立大鼠肾纤维化模型,动态观察4周。治疗的第12、、3周末检测血肌酐、尿素氮含量等指标,观察肾功能变化;4周末采用HE染色、六胺银(periodic acid-silver methenamine,PASM)染色和丽春红染色观察肾组织病理变化。结果模型组大鼠血肌酐、尿素氮均有明显上升;模型组大鼠大部分肾小球呈玻璃样变,硬化的肾小球周围所属肾小管萎缩、基底膜增厚,部分肾小管消失;少数残存的肾小球肥大并周围肾小管扩张严重;肾间质胶原纤维增生和大量炎细胞浸润。结论该模型有明显的肾间质纤维化特征,且死亡率低,适合肾间质纤维化的实验研究。     

依帕司他对单侧输尿管梗阻大鼠肾间质纤维化的干预作用及其机制

目的:观察依帕司他(EPS)对单侧输尿管梗阻(UUO)大鼠间质纤维化的保护作用及其机制。方法:实验设假手术组(Sham)组、UUO、UUO+EPS(50 mg/kg)及UUO+EPS(100 mg/kg)剂量组,每组n=8。左侧输尿管结扎制备UUO大鼠模型。造模后连续灌胃给药3周,sham和UUO组给予等体积的羟甲基纤维素钠。HE和Masson染色观察肾组织病理变化及胶原沉积情况。免疫组化法观察肾组织醛糖还原酶(AR)表达情况,分别采用real-time PCR和(或) Western blot检测肾脏I型胶原(collagen I)、III型胶原(collagen III)、α-平滑肌肌动蛋白(α-SMA)、成纤维细胞特异蛋白-1(FSP-1)、纤连蛋白(FN)、E-钙粘蛋白(E-cadherin)、转化生成因子-β1(TGF-β1)和AR mRNA及蛋白表达。结果:与Sham组相比,UUO组大鼠小管上皮细胞萎缩、空泡样变性,肾间质成纤维细胞及肌成纤维细胞大量增殖并伴大量炎症细胞浸润,胶原沉积明显增加,collagen I、collagen III、TGF-β1和AR mRNA及蛋白表达水平明显升高(P<0.01),同时EMT标志性蛋白α-SMA、FSP-1、FN mRNA及蛋白表达水平明显升高(P<0.01),而E-cadherin mRNA及蛋白表达水平明显降低。与UUO组相比,经EPS治疗3周后,肾间质纤维化程度明显减轻,胶原沉积明显减少,collagen I、collagen III、TGF-β1和AR mRNA及蛋白表达水平明显降低(P<0.01或P<0.05),另外α-SMA、FSP-1、FN mRNA及蛋白表达水平明显降低(P<0.01或P<0.05),而E-cadherin mRNA及蛋白表达水平明显升高(P<0.01或P<0.05),而且100 mg/kg剂量组上述指标的改变均好于低剂量组(P<0.05,P<0.01)。结论:依帕司他对肾间质纤维化具有一定的改善作用,其机制可能与其抑制TGF-β1介导的AR表达、进而抑制大鼠肾小管上皮细胞EMT有关。     

甲磺酸伊马替尼对单侧输尿管梗阻小鼠肾间质纤维化的保护作用

目的研究甲磺酸伊马替尼（STI571）改善单侧输尿管梗阻（UUO）小鼠肾间质纤维化的作用及机制。方法48只小鼠随机分为4组：假手术组,模型组,小剂量治疗组（80mg/kg/d）,大剂量治疗组（160mg/kg/d）。采用左侧输尿管双结扎的方法建立UUO模型,治疗组每天以STI57180、160mg/Kg灌胃。分别于术后第8,11d分别处死各组小鼠6只。光镜下观察肾脏病理改变。用免疫组化技术检测肾组织TGF-β1、PAI-1、α-SMA和PCNA的表达。结果治疗组的肾间质纤维化定量显著低于模型组（P〈0.05）,且不同剂量组之间存在显著差异（P〈0.05）。模型组和治疗组左肾TGF-β1、PAI-1、α-SMA和PCNA的表达均随梗阻时间延长而逐渐增加,治疗组α-SMA和PCNA的表达较模型组明显减低（P〈0.05）。结论甲磺酸伊马替尼可显著减轻UUO小鼠梗阻侧肾脏间质纤维化,下调α-SMA和PCNA的表达,减少肾间质细胞外基质的沉积,对UUO小鼠肾间质纤维化有一定防治作用。     

口服氯化汞对大鼠肾间质纤维化的作用

目的口服氯化汞(HgCl2)造成大鼠的肾间质纤维化模型并探讨相关机制。方法用不同剂量HgCl2[(A组为5mg/(kg·bw)、B组为10mg/(kg·bw)、C组为20mg/(kg·bw)]给大鼠灌胃1周,观察大鼠一般状况、肾功能和肾组织病理变化,免疫组化法观察肾组织纤维连接蛋白(FN)和α-平滑肌肌动蛋白(α-SMA)表达。结果模型大鼠体重下降,肾体比增加,肾功能损害和肾组织Hyp含量呈剂量依赖性升高,肾间质炎性细胞浸润,肾间质胶原沉积增加,肾间质FN和α-SMA表达增强,以C组病变最重。结论20mg/(kg·bw)剂量HgCl2灌胃1周可造成大鼠的肾间质纤维化病变,其部分机制在于HgCl2促使肾间质肌成纤维细胞活化和细胞外基质的生成沉积。     

上皮—间质转化在肾间质纤维化中的作用

上皮－间质转化在发育和纤维化过程中具有重要作用。本文综述了上皮－间质转化发生的过程及其机制的研究进展,尤其是细胞外基质、生长因子、粘附分子及基因对上皮－间质转化的影响。并就在肾间质纤维化过程中因上皮－间质转化致成纤维细胞增多,从而导致肾纤维化的可能作用及其影响因素作一述。     

TNFRSF13C 在单侧输尿管梗阻大鼠肾组织中的表达及其意义

目的：检测单侧输尿管梗阻（UUO）大鼠肾组织中B 细胞激活因子受体（TNFRSF13C）的表达变化,探讨其在肾间质纤维化 病变中的作用。方法：采用UUO法建立肾间质纤维化大鼠模型,20只成年雄性大鼠,随机分为4组,分别于术后0、3、7、14 天处死 大鼠。取左侧梗阻肾脏进行Masson染色,拍照后,采用双盲法评定各组肾小管间质纤维化程度。提取肾组织中总RNA,用实时荧 光定量聚合酶链反应（RT-PCR）法检测各组肾组织中TNFRSF13C基因表达情况。Pearson 检测TNFRSF13C表达量与肾小管间质 纤维化程度的相关性。结果：随着梗阻时间的延长,肾组织中TNFRSF13C 的mRNA 表达量进行性升高,与肾间质纤维化病变程 度一致,两者呈显著正相关（r=0.915,P<0.01）。结论：TNFRSF13C可能在肾间质纤维化病程中起到了重要作用,并有望成为慢性 肾脏病的临床监测指标。     

肾小球内皮细胞-间质转化在糖尿病肾病肾纤维化中的作用

肾纤维化是糖尿病肾病(diabetic kidney disease,DKD)重要的病理特征。肾小球内皮细胞-间质转化(endothelial-to-mesenchymal transition,EndoMT)可促进肾纤维化,在DKD的发生发展中起重要作用,但其具体机制仍有待研究。本文就EndoMT在DKD肾纤维化中的分子机制以及目前通过抗EndoMT治疗延缓DKD肾纤维化的研究进展作一综述,以期为DKD的临床治疗提供新的理论依据。     

沉默Wnt4基因抑制大鼠肾间质纤维化

目的探讨沉默Wnt4基因对肾间质纤维化的影响,为慢性肾病的治疗提供理论依据。方法 128只SD大鼠随机分为假手术组、模型组、阴性对照组和Wnt4基因沉默组,每组32只。构建Wnt4 siRNA慢病毒载体体内转染沉默组的大鼠,于转染后第3、7、10、14天分为四个亚组,每组8只大鼠。通过H&E染色病理检查、RT-PCR技术检测肾间质改变及β-连环蛋白、Wnt4、α-SMA表达情况。结果 H&E染色病理检查结果表明:假手术组四个时间点未见肾间质改变;UUO组、阴性沉默组及沉默组造模后3 d出现肾间质水肿、少量肾间质纤维化,10、14 d肾间质弥漫性巨噬细胞、淋巴细胞浸润,呈加重趋势;阴性沉默组基因与UUO组相同;沉默组四个时间点均出现不同程度肾间质纤维化,14 d肾间质纤维化程度低于阴性沉默组及UUO组(P<0.05);Wnt4基因沉默后UUO组mRNA表达量的相关性分析显示,Wnt4与β-catenin mRNA表达量具有显著相关性(r=0.886,P<0.001)。Wnt4与α-SMA mRNA表达量具有显著相关性(r=0.930,P<0.001)。Wnt4基因沉默后沉默组mRNA表达量的相关性分析显示,Wnt4与β-catenin mRNA表达量无显著相关性(r=0.204,P=0.263)。Wnt4与α-SMA mRNA表达量具有显著相关性(r=0.753,P<0.001)。结论沉默Wnt4基因在肾间质纤维化大鼠中可明显抑制肾间质纤维化,可能对其有治疗作用。     

肾间质纤维化相关信号通路的研究进展

肾间质纤维化是以正常的肾间质和肾小管结构被大量聚集的细胞外基质所替代为特征的病理过程,是多数慢性肾脏疾病进展为终末期肾衰竭共同的病变过程,其病理变化主要由多种细胞因子和多条信号通路控制,是众多关键信号通路的交互影响与共同作用的结果。深入了解信号通路的相互作用对进一步揭示肾间质纤维化的分子机制有重要意义。现综述肾间质纤维化病理变化中关键的信号通路,以期为肾间质纤维化分子机制的研究提供参考。     

microRNA与肾间质纤维化的研究进展

小分子核糖核酸(microRNA)是一类约20个核苷酸单链,在转录后水平调节基因的表达。microRNA广泛分布于人体各个组织器官内,但同时也有显著的组织特异性,不同的组织器官中miRNA的表达强度有显著差异,某些microRNAs在肾脏组织中呈特异性的高表达。肾间质纤维化是各种慢性肾脏病进展至终末期,最终导致器官功能丢失的共同的病理过程和特征。通过多年累积的研究表明,一些特定的microRNAs与肾间质纤维化的进程密切相关,在这个过程中体现出极其复杂的调控机制,发挥多方面的作用。近年来,随着对microRNA的研究进一步深入,本文就microRNAs在肾间质纤维化进程中的表达特点、作用靶点及相关调控机制的研究进展进行如下综述。     

microRNA与肾间质纤维化的研究进展

小分子核糖核酸（microRNA）是一类约20个核苷酸单链,在转录后水平调节基因的表达。microRNA广泛分布于人体各个组织器官内,但同时也有显著的组织特异性,不同的组织器官中miRNA的表达强度有显著差异,某些microRNAs在肾脏组织中呈特异性的高表达。肾间质纤维化是各种慢性肾脏病进展至终末期,最终导致器官功能丢失的共同的病理过程和特征。通过多年累积的研究表明,一些特定的microRNAs与肾间质纤维化的进程密切相关,在这个过程中体现出极其复杂的调控机制,发挥多方面的作用。近年来,随着对microRNA的研究进一步深入,本文就microRNAs在肾间质纤维化进程中的表达特点、作用靶点及相关调控机制的研究进展进行如下综述。     

Amniotic Fluid Derived Stem Cells with a Renal Progenitor Phenotype Inhibit Interstitial Fibrosis in Renal Ischemia and Reperfusion Injury in Rats

Objectives

Mesenchymal stem cells derived from human amniotic fluid (hAFSCs) are a promising source for cellular therapy, especially for renal disorders, as a subpopulation is derived from the fetal urinary tract. The purpose of this study was to evaluate if hAFSCs with a renal progenitor phenotype demonstrate a nephroprotective effect in acute ischemia reperfusion (I/R) model and prevent late stage fibrosis.

Methods

A total of 45 male 12-wk-old Wistar rats were divided into three equal groups;: rats subjected to I/R injury and treated with Chang Medium, rats subjected to I/R injury and treated with hAFSCs and sham-operated animals. In the first part of this study, hAFSCs that highly expressed CD24, CD117, SIX2 and PAX2 were isolated and characterized. In the second part, renal I/R injury was induced in male rats and cellular treatment was performed 6 hours later via arterial injection. Functional and histological analyses were performed 24 hours, 48 hours and 2 months after treatment using serum creatinine, urine protein to creatinine ratio, inflammatory and regeneration markers and histomorphometric analysis of the kidney. Statistical analysis was performed by analysis of variance followed by the Tukey’s test for multiple comparisons or by nonparametric Kruskal-Wallis followed by Dunn. Statistical significance level was defined as p <0.05.

Results

hAFSCs treatment resulted in significantly reduced serum creatinine level at 24 hours, less tubular necrosis, less hyaline cast formation, higher proliferation index, less inflammatory cell infiltration and less myofibroblasts at 48h. The treated group had less fibrosis and proteinuria at 2 months after injury.

Conclusion

hAFSCs contain a renal progenitor cell subpopulation that has a nephroprotective effect when delivered intra-arterially in rats with renal I/R injury, and reduces interstitial fibrosis on long term follow-up.     

BMP-7在抑制肾脏纤维化中的作用

肾脏纤维化过程十分复杂,受到多种因素的作用,但总体上可以归结为两方面,一方面是促进纤维化的因素,为正调节因素;另一方面是抗纤维化的因素,为负调节因素。近年来,发现了一些肾脏纤维化的内源性负调节因素,比较公认的有肝细胞生长因子、骨形态发生蛋白、核心蛋白聚糖和过氧化物酶体增殖物激活受体等。该文介绍骨形态发生蛋白-7（BMP-7）在抑制肾脏纤维化方面的研究进展。     

Effect of Interleukin 6 Deficiency on Renal Interstitial Fibrosis

Our recent studies have shown that bone marrow-derived fibroblast precursors contribute significantly to the pathogenesis of renal fibrosis. However, the molecular mechanisms underlying the recruitment and activation of bone marrow-derived fibroblast precursors are incompletely understood. We found that interleukin 6 was induced in the kidney in a murine model of renal fibrosis induced by unilateral ureteral obstruction. Therefore, we investigated if interleukin 6 play a role in the recruitment and maturation of bone marrow-derived fibroblast precursors in the kidney during the development of renal fibrosis. Wild-type and interleukin 6 knockout mice were subjected to unilateral obstructive injury for up to two weeks. Interleukin 6 knockout mice accumulated similar number of bone marrow-derived fibroblast precursors and myofibroblasts in the kidney in response to obstructive injury compared to wild-type mice. Furthermore, IL-6 knockout mice expressed comparable α-SMA in the obstructed kidney compared to wild-type mice. Moreover, targeted disruption of Interleukin 6 did not affect gene expression of profibrotic chemokine and cytokines in the obstructed kidney. Finally, there were no significant differences in renal interstitial fibrosis or expression of extracellular matrix proteins between wild-type and interleukin 6 knockout mice following obstructive injury. Our results indicate that interleukin 6 does not play a significant role in the recruitment of bone marrow-derived fibroblast precursors and the development of renal fibrosis.     

羟苯磺酸钙通过抑制Ⅰ型胶原表达减轻肾间质纤维化

目的:研究羟苯磺酸钙对小鼠肾间质纤维化、Ⅰ型胶原表达的影响。方法:将C57小鼠随机分为假手术组(Sham组,n=4)、肾间质纤维化模型组(UUO组,n=5)及羟苯磺酸钙治疗组(CDT组,n=4);采用单侧输尿管梗阻制备肾间质纤维化模型,CDT组给予羟苯磺酸钙灌胃、Sham组和UUO组给予双蒸水灌胃;采用HE染色、Masson染色、免疫组化、实时定量PCR以及蛋白免疫印迹观察单侧输尿管梗阻术后14 d小鼠术侧肾脏的肾间质纤维化程度和Ⅰ型胶原表达情况。结果:与Sham组比较,UUO组小鼠术后14 d术侧肾脏肾发生显著肾间质纤维化,Ⅰ型胶原表达显著增强(Ⅰ型胶原基因相对表达量:Sham组:1.00000,UUO组:114.92289,P0.0001)。与UUO组比较,CDT组小鼠术后14 d术侧肾间质纤维化程度显著减轻,Ⅰ型胶原表达显著减弱(Ⅰ型胶原基因相对表达量:UUO组:114.92289,CDT组:45.33516,P0.005)。结论:羟苯磺酸钙通过抑制小鼠肾间质Ⅰ型胶原表达从而减轻单侧输尿管结扎小鼠肾间质纤维化。     

和络泄浊颗粒对大鼠肾间质纤维化miR-200a表达影响

目的：探索和络泄浊颗粒对大鼠肾脏纤维化中miR-200a 表达的影响及探讨其可能存在的作用机制。方法：30 只SD雄性大鼠随机分为6 组：假手术组(Sham)、手术组(UUO)及和络泄浊颗粒(UUO+ REG)组各2 组,运用单侧（左）输尿管结扎法制造肾间质纤维化模型,但Sham组仅游离输尿管,而其余两组则游离并结扎输尿管。术后各组按1 mg/kg·d-1的量进行灌胃,UUO+ REG组给予REG,其余两组则予生理盐水。术后第7 天和14 天,摘除左梗阻侧肾脏进行免疫组化检查alpha-SMA和荧光定量PCR 检测miR-200a 的表达。结果：在UUO及UUO+ REG 组,alpha-SMA 表达明显高于Sham 组（P＜0.01）;UUO+ REG 组低于UUO 组（P＜0.05）。miR-200a 表达水平在UUO 组和UUO+ REG 组都是降低的,但是UUO 组与Sham 组差别明显（P＜0.01）,其与UUO+REG组亦有明显的差别（P＜0.05）。结论：和络泄浊颗粒可以通过上调miR-200a 延缓肾脏纤维化进展。     

Proteinuria Triggers Renal Lymphangiogenesis Prior to the Development of Interstitial Fibrosis

Proteinuria is an important cause of progressive tubulo-interstitial damage. Whether proteinuria could trigger a renal lymphangiogenic response has not been established. Moreover, the temporal relationship between development of fibrosis, inflammation and lymphangiogenesis in chronic progressive kidney disease is not clear yet. Therefore, we evaluated the time course of lymph vessel (LV) formation in relation to proteinuria and interstitial damage in a rat model of chronic unilateral adriamycin nephrosis. Proteinuria and kidneys were evaluated up to 30 weeks after induction of nephrosis. LVs were identified by podoplanin/VEGFR3 double staining. After 6 weeks proteinuria was well-established, without influx of interstitial macrophages and myofibroblasts, collagen deposition, osteopontin expression (tubular activation) or LV formation. At 12 weeks, a ∼3-fold increase in cortical LV density was found (p<0.001), gradually increasing over time. This corresponded with a significant increase in tubular osteopontin expression (p<0.01) and interstitial myofibroblast numbers (p<0.05), whereas collagen deposition and macrophage numbers were not yet increased. VEGF-C was mostly expressed by tubular cells rather than interstitial cells. Cultured tubular cells stimulated with FCS showed a dose-dependent increase in mRNA and protein expression of VEGF-C which was not observed by human albumin stimulation. We conclude that chronic proteinuria provoked lymphangiogenesis in temporal conjunction with tubular osteopontin expression and influx of myofibroblasts, that preceded interstitial fibrosis.     

Epigallocatechin-3-Gallate Attenuates Unilateral Ureteral Obstruction-Induced Renal Interstitial Fibrosis in Mice

The severity of tubulointerstitial fibrosis is regarded as an important determinant of renal prognosis. Therapeutic strategies targeting tubulointerstitial fibrosis have been considered to have potential in the treatment of chronic kidney disease. This study aims to evaluate the protective effects of (-)-epigallocatechin-3-gallate (EGCG), a green tea polyphenol, against renal interstitial fibrosis in mice. EGCG was administrated intraperitoneally for 14 days in a mouse model of unilateral ureteral obstruction (UUO). The results of our histological examination showed that EGCG alleviated glomerular and tubular injury and attenuated renal interstitial fibrosis in UUO mice. Furthermore, the inflammatory responses induced by UUO were inhibited, as represented by decreased macrophage infiltration and inflammatory cytokine production. Additionally, the expression of type I and III collagen in the kidney were reduced by EGCG, which indicated an inhibition of extracellular matrix accumulation. EGCG also caused an up-regulation in α-smooth muscle actin expression and a down-regulation in E-cadherin expression, indicating the inhibition of epithelial-to-mesenchymal transition. These changes were found to be in parallel with the decreased level of TGF-β1 and phosphorylated Smad. In conclusion, the present study demonstrates that EGCG could attenuate renal interstitial fibrosis in UUO mice, and this renoprotective effect might be associated with its effects of inflammatory responses alleviation and TGF-β/Smad signaling pathway inhibition.