The role of nitric oxide in reflux nephropathy

Reflux nephropathy (RN) is recognized as a major cause of end-stage renal failure in children and young adults. Inhibition of nitric oxide (NO) exacerbates and enhanced production ameliorates tubulointerstitial fibrosis (TIF) in experimental obstructive uropathy. NO is synthesised by NO synthase (NOS), three distinct isoforms of which have been identified: inducible (iNOS), endothelial (eNOS), and neuronal (nNOS). It has been reported that iNOS induces immunologic injury to glomerular cells and enhances accumulation of extracellular matrix in the glomerulus and tubulointerstitial space. Furthermore, it has been suggested that nNOS and eNOS have beneficial effects in ameliorating TIF. We investigated the expression of different isoforms of NOS in severe refluxing kidneys in order to further understand the pathogenesis of RN in kidney specimens from nine children with severe RN obtained at nephrectomy. Control material included normal kidney specimens from three adult patients undergoing partial nephrectomy for small kidney tumours. Histochemistry for NO was performed using nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase. Single-label immunofluorescence histochemistry was carried out using polyclonal antibodies to nNOS, iNOS, eNOS, and transforming growth factor (TGF)-beta 1 employing laser-scanning confocal microscopy. The TUNEL method was used to assess tubular apoptosis. Strong NADPH staining was observed in the proximal tubules of RN kidneys compared to controls, where there was weak staining. Control kidneys demonstrated weak immunoreactivity for iNOS in the proximal tubules and a lack of immunoreactivity for nNOS and eNOS. RN kidneys demonstrated strong immunoreactivity for nNOS in the tubulointerstitial space, for eNOS in the glomerulus, and for iNOS in the glomerulus and proximal tubules. Strong immunoreactivity for TGF beta 1 was seen in the glomerulus and proximal tubules identical to iNOS. Increased immunoreactivity for iNOS and TGF-beta 1 strongly correlated with the severity of apoptosis in RN. Our data demonstrate that NO derived from nNOS, iNOS, and eNOS is strongly expressed in RN. The selective shunting of NO via iNOS may induce renal fibrosis in RN. The upregulation of nNOS and eNOS in RN appears to be a compensatory mechanism of ameliorating TIF.     

牛初乳短链胰岛素样生长因子-1对多柔比星肾病大鼠胰岛素样生长因子-1的影响

目的研究牛初乳短链胰岛素样生长因子-1（BctIGF-1）对肾病大鼠胰岛素样生长因子-1（IGF-1）的影响,为治疗肾病综合征（NS）提供理论依据。方法将30只周龄相同、体质量相近的雌性SD大鼠随机分为对照组、肾病组、BctIGF-1治疗组;实验第6周末杀鼠,采用放射免疫法测定其血清与尿IGF-1水平,反转录酶-聚合酶链反应（RT-PCR）测定其肝、肾组织IGF-1 mRNA表达,采用考马斯亮蓝法测定24h尿蛋白排泄量。结果BctIGF-1治疗组24h尿蛋白及病理积分均低于肾病组（P〈0.05,0.01）;肾病大鼠血清IGF-1水平明显低于BctIGF-1治疗组（P〈0.05）,尿IGF-1排泄量明显高于BctIGF-1治疗组（P〈0.01）;各组大鼠肝组织IGF-1 mRNA表达差异无显著性;肾组织IGF-1mRNA表达由高至低依次为对照组、BctIGF-1治疗组、肾病组,BctIGF-1治疗与对照组比较差异无显著性,但BctIGF-1治疗与对照组IGF-1 mRNA表达均显著高于肾病组（P〈0.01,0.05）。结论BctIGF-1可有效降低肾病大鼠尿蛋白,减轻肾脏病理损伤,其机制可能是通过增加其血清游离IGF-1水平、促进肾局部IGF-1mRNA表达来实现。     

Microdissection demonstration of the lesion of the endocrine kidney in children

Microdissection of nephrons of kidneys of children showing advanced ischemic tubular atrophy, and removed for control of hypertension, demonstrates marked proximal convoluted tubular atrophy, with formation of multiple small proximal tubular diverticula. These diverticula presumably contribute to the microscopic appearance of large numbers of small tubules lined by low epithelial cells with pale or clear cytoplasm, adjacent to glomeruli in the cortices of kidneys showing ischemic tubular atrophy (endocrine kidney). Segmentation of such atrophic tubules leads to formation of blind segments (microcysts), as demonstrated in this study and by Oliver. The distinctive microscopic appearance of the endocrine kidney, a not infrequent finding in kidneys of children with chronic renal insufficiency who require nephrectomy for control of hypertension, has not hitherto been emphasized in the literature on pediatric renal disease.     

Microdissection Demonstration of the Lesion of the Endocrine Kidney in Children

Microdissection of nephrons of kidneys of children showing advanced ischemic tubular atrophy, and removed for control of hypertension, demonstrates marked proximal convoluted tubular atrophy, with formation of multiple small proximal tubular diverticula. These diverticula presumably contribute to the microscopic appearance of large numbers of small tubules lined by low epithelial cells with pale or clear cytoplasm, adjacent to glomeruli in the cortices of kidneys showing ischemic tubular atrophy (endocrine kidney). Segmentation of such atrophic tubules leads to formation of blind segments (microcysts), as demonstrated in this study and by Oliver.⁸ The distinctive microscopic appearance of the endocrine kidney, a not infrequent finding in kidneys of children with chronic renal insufficiency who require nephrectomy for control of hypertension, has not hitherto been emphasized in the literature on pediatric renal disease.     

Effects of maternal subtotal nephrectomy on the development of the fetal kidney: A morphometric study

The present study was designed to explore if maternal subtotal (5/6) nephrectomy affects the development of fetal rat kidneys using morphometric methods and examining whether there are any apoptotic changes in the fetal kidney. To generate 5/6 nephrectomized model rats, animals underwent 2/3 left nephrectomy on gestation day (GD) 5 and total right nephrectomy on GD 12. The fetal kidneys were examined on GDs 16 and 22. A significant decrease in fetal body weight resulting from maternal 5/6 nephrectomy was observed on GD 16, and a significant decrease in fetal renal weight and fetal body weight caused by maternal nephrectomy was observed on GD 22. Maternal 5/6 nephrectomy induced a significant increase in glomerular number, proximal tubular length, and total proximal tubular volume of fetuses on GD 22. Maternal 5/6 nephrectomy resulted in an increase in the number of apoptotic cells in the metanephric mesenchyme of the kidney on GD 16, and in the collecting tubules on GD 22. These findings suggest that maternal 5/6 nephrectomy stimulates the development of the fetal kidney while suppressing fetal growth.     

Cellular response to renal hypoxia is different in adolescent and infant rats

Immature renal tubules are more tolerant to ischemia than mature renal tubules. Here we compared the developmental pattern for some cellular responses evoked by hypoxia and reoxygenation in renal proximal tubules from 10- and 40-day-old rats. Redistribution of Na(+)-K(+)-ATPase from the plasma membrane was studied by confocal microscopy techniques in primary cultured renal proximal tubular cells. The developmental expression of Na(+)-K(+)-ATPase, micro-calpain and heme oxygenase-1 was measured by RT-PCR techniques in rat renal cortex. In response to hypoxia Na(+)-K(+)-ATPase redistribution from the plasma membrane was almost 2-fold increased in cells isolated from mature kidneys compared with cells isolated from immature kidneys. Reoxygenation resulted in a complete reestablishment of Na(+)-K(+)-ATPase in the plasma membrane in the immature but not in the mature cells. The dissociation of Na(+)-K(+)-ATPase from the plasma membrane was associated with a reduced activity and a reduced expression of Na(+)-K(+)-ATPase in the mature but not in the immature tubular cells. The expression of micro-calpain, a factor shown to induce ischemic injury to proximal tubular cells, was significantly lower in the immature compared with the mature kidney, whereas the expression of heme oxygenase-1, a factor shown to protect from renal ischemic injury, was significantly higher in the immature kidney. The results help to explain the increased tolerance of the immature kidney to injury caused by ischemia and reperfusion.     

胰岛素样生长因子-1(IGF-1)对缺氧缺血脑损伤新生鼠内源性IGF-1和IGF-1受体基因表达的影响

目的　胰岛素样生长因子 1(IGF 1)对损伤的神经组织有修复作用 ,但外源性IGF 1是否会抑制内源性IGF 1、IGF 1受体的生成 ,从而减弱IGF 1的神经保护作用尚不明确。本文通过观察IGF 1治疗新生大鼠缺血缺氧脑损伤 (HIBD)后脑IGF 1和IGF 1受体mRNA水平的变化 ,研究IGF 1对HIBD新生大鼠内源性IGF 1、IGF 1受体的影响。方法　制作新生大鼠HIBD模型 ,用原位杂交方法观察HIBD后各时间点海马和大脑皮层IGF 1和IGF 1受体基因表达的动态变化 ,并比较IGF 1治疗组与未治疗组HIBD后 12h、72hIGF 1、IGF 1受体mRNA的表达水平。结果　HIBD后 4 8h海马IGF 1和IGF 1受体mRNA开始升高 ,72h达高峰。损伤后 12 0h ,IGF 1mRNA降至正常水平 ,而IGF 1受体mRNA仍处于较高水平。在皮层 ,IGF 1和IGF 1受体mRNA开始升高时间稍早于海马 ,2 4h上升 ,96h降至正常 ,但是上升幅度相对较小。与未治疗组比较 ,IGF 1治疗后内源性IGF 1表达无明显变化。IGF 1受体的表达在治疗后 12h无明显差别 ,但在 72h时显著增加。结论　HIBD后皮层、海马等脑损伤区的IGF 1和IGF 1受体表达均升高。给予外源性IGF 1后并不降低内源性IGF 1的表达 ,还能刺激IGF 1受体表达增加。     

百令胶囊对肾小管间质纤维化大鼠肾小管上皮-间质转分化的干预作用

目的探讨百令胶囊对小管间质纤维化大鼠小管上皮-间质转分化的干预作用。方法利用腺嘌呤诱导建立肾小管间质纤维化大鼠模型,实验SD大鼠随机分为模型组、干预组、对照组各30只,分别于实验第7周、12周、17周收获动物各10只,进行功能学和肾脏组织病理学检测和观察。利用免疫组织化学对骨形态发生蛋白-7(BMP-7)、转化生长因子-β1(TGF-β1)和α-平滑肌肌动蛋白(-αSMA)在肾小管间质纤维化大鼠中的表达变化进行动态观察及百令胶囊的干预影响。结果实验第7周模型组大鼠即表现出大量蛋白尿、小管损伤、间质的轻度纤维化和炎症细胞浸润(P<0.01);随病程进展,上述病变呈进行性加重(P<0.01)。百令胶囊干预组动物在实验第7周、12周功能学组织学的改善同实验组相比有明显差异(P<0.01),17周后二者无显著差异。免疫组织化学显示百令胶囊12周前能显著上调BMP-7的表达和降低-αSMA和TGF-β1在肾脏小管间质中的表达(P<0.01),12周后这种变化无差异。结论百令胶囊在发病早期通过有效干预上皮-间质转分化达到改善肾脏纤维化的作用,随着肾小管间质纤维化程度的加重,百令胶囊逐渐失去其阻断作用。     

IGF-IR单克隆抗体对Jurkat细胞增殖及凋亡的影响

目的探讨胰岛素样生长因子Ⅰ受体(IGF-IR)在急性T淋巴细胞白血病(T-ALL)细胞系Jurkat的表达、定位,观察IGF-IR单克隆抗体(MAb)对Jurkat细胞增殖、凋亡的影响。方法 (1)采用免疫细胞化学的方法检测IGF-IR在Jurkat细胞的表达、定位。(2)分别用5个不同实验浓度:0.001μg/mL,0.01μg/mL,0.1μ/mL,1μg/mL,10μg/mL的IGF-IR MAb作用Jurkat细胞48h,用CCK-8试剂盒检测细胞的增殖抑制率。(3)选用10μg/mL的IGF-IR MAb作用Jurkat细胞48h,上流式细胞仪测定细胞的凋亡情况。结果 (1)IGF-IR在Jurkat细胞株100%表达,主要为细胞膜、细胞浆混合表达。(2)0.001μg/mL,0.01μg/mL,0.1μg/mL,1μg/mL,10μg/mL的IGF-IR MAb作用Jurkat细胞株48 h,Jurkat细胞的增殖抑制率分别为:(9.67±1.17)%,(14.89±1.06)%,(17.64±0.81)%,(20.15±1.14)%,(24.10±1.11)%,各组间进行两两比较,差异有显著性(P0.05)。(3)10μg/mL的IGF-IR MAb作用Jurkat细胞48h,实验组的诱导细胞凋亡率分别为:早期凋亡率(13.73±1.16)%,晚期凋亡率(20.44±2.47)%,总凋亡率(34.18±3.41)%;对照组的诱导细胞凋亡率分别为:早期凋亡率(6.27±0.67)%,晚期凋亡率(10.18±0.81)%,总凋亡率(16.45±1.38)%。实验组细胞的早期凋亡率,晚期凋亡率,总凋亡率均较对照组高,差异有显著性(P0.05)。结论 (1)Jurkat细胞普遍表达IGF-IR,主要为细胞膜、细胞浆混合表达。(2)IGF-IR MAb可使Jurkat细胞株的增殖受到抑制,且IGF-IR MAb的浓度越高,抑制率越大。(3)IGF-IR MAb可使Jurkat细胞的凋亡增加。     

Labeled Lectin Studies of Renal Tubular Dysgenesis and Renal Tubular Atrophy of Postnatal Renal Ischemia and end-Stage Kidney Disease

Renal tubular dysgenesis (RTD), with hypoplasia especially of renal proximal convoluted tubules and clinical neonatal anuria or oliguria, has been reported as a congenital familial (autosomal recessive) disease, variably with features of oligohydramnios, Potter syndrome, or pulmonary hypoplasia. A similar tubular lesion due to antenatal tubular atrophy has been reported for conjoined twins with twin-twin transfusion syndrome or acardia and in infants of mothers given antihypertensive agents, including angiotensin-converting enzyme (ACE) inhibitors, during pregnancy, and it has been seen as a unilateral lesion in young infants with renal artery stenosis due to arteritis or medial arterial calcinosis. The renal tubular changes in RTD are very like those of the “endocrine kidney” in experimental animals and resemble those of the renal tubular atrophy of end-stage kidney diseases such as glomerulonephritis, tubulointerstitial kidney disease, obstructive uropathy/pyelonephritis, graft rejection of transplanted kidneys, or the renal parenchymal changes seen with protracted dialysis therapy. Labeled lectins that differentially mark proximal convoluted, distal convoluted and connecting, and collecting tubules showed no distinctive differences in stainingpatterns of the hypoplastic renal tubules of infants and children with RTD, postnatal renal artery obstruction, or the various types of end-stage renal disease with the lectins used (PNA, GSL1, UEA, and LTA). The findings suggest that the renal tubular changes in some if not all the conditions studied are the result of renal ischemia. The reported familial RTD with hypernephronic nephromegaly may be a specific disorder, but other forms could reflect renal ischemia acquired in utero or in early or later postnatal life.     

蛋白尿负荷幼鼠肾组织核转录因子κB、致纤维化因子及纤维连接蛋白核酸表达的研究

目的探讨幼年大鼠持续蛋白尿致肾损伤的不同时间点肾组织NF-κB亚单位P65/Rel-A及凝血酶敏感蛋白(TSP-1)、转化生长因子(TGF-β1)、结缔组织生长因子(CTGF)和纤维连接蛋白(FN)mRNA动态表达的趋势。方法3周龄幼年W istar雌性大鼠80只,分为BSA组(40只)和对照组(40只),采用BSA诱导制备蛋白负荷肾病模型;考马斯亮蓝比色测大鼠尿蛋白;HE染色评价肾组织常病理变化;原位杂交检测P65/Rel-A、TSP-1、TGF-β1及CTGF mRNA表达;Northern b lot检测FNmRNA表达。SPSS10对实验数据进行统计学处理。结果(1)BSA组大鼠至3~4周蛋白尿达大量蛋白尿程度,肾间质炎症细胞浸润,肾小管蛋白管型形成,间质水肿,间质区加宽。(2)BSA组各级肾小管上皮细胞P65/Rel-A mRNA于细胞核的表达强度趋于增加,第1、2、3、4周半定量积分分别为:2.33±0.20、2.76±0.12、2.96±0.19、3.76±0.18(F=37.34,P<0.01)。(3)BSA组TSP-1 mRNA表达第1、2、3、4周半定量积分分别为:2.60±0.28、3.39±0.41、2.77±0.08、2.71±0.13,于第2周时达高峰(F=11.14,P<0.01)。(4)伴随蛋白尿的加重,BSA组TGF-β1及CTGF的mRNA于各级肾小管上皮细胞表达趋势进行性增强,与对照组比较及自身各时间点比较差异均有统计学意义(P<0.01)。(5)Northern b lot结果提示,BSA组FN mRNA于第2周明显上调,至第4周是对照组的3.6倍,是第1周的2.7倍,与对照组比较差异均有统计学意义(P<0.01)。结论大量持续蛋白尿过程中肾组织NF-κB信号途径活化,TSP-1、TGF-β1、CTGF的异常表达,肾间质FN的合成和异常集聚,可能是促进肾间质进一步损伤的机制之一。     

小儿肾积水尿细胞因子IGF-1、ET-1、TGF-β1及MCP-1水平的检测及意义

目的 探讨尿细胞因子(胰岛素样生长因子IGF-1、内皮素ET-1、转化生长因子TGF-β及单核细胞趋化蛋白MCP-1)的检测在判定小儿肾积水病肾损害程度中的临床意义.方法 采用酶联免疫吸附法(ELISA)检测41例小儿肾积水病肾尿IGF-1、ET-1、TGF-β1>及MCP-1水平,并以健肾尿作对照.41例同时行病肾组织学检查分级,并与病肾尿IGF-1、ET-1、TGF-β1>及MCP-1水平作相关分析.结果 ①病肾尿IGF-1明显降低为(186.69±24.63)pg/ml,健肾测值为(279.45±31.57)pg/ml,差异有统计学意义(P<0.01);②病肾尿ET-1、TGF-β1、MCP-1明显升高,分别为(49.81±9.08)、(395.91±83.52)、(486.59±89.72)pg/ml,健肾相应测值为(13.21±2.91)、(232.57±32.68)、(328.54±36.81)pg/ml,差异均有统计学意义(P<0.05);③病肾尿IGF-1与病理分级密切负相关,差异有统计学意义(r=-0.839,P<0.01);ET-1、TGF-β1及MCP-1水平与病理分级正相关,但差异无统计学意义(P>0.05).结论 小儿肾积水病肾尿IGF-1水平是从尿细胞因子检测中判定病肾损害程度的理想指标之一.     

胰岛素样生长因子-1受体基因与特发性矮小相关性研究进展

生长激素-胰岛素样生长因子(GH-IGF)轴是调节儿童生长发育中最重要的神经内分泌轴.胰岛素样生长因子-1受体( IGF-1 R)是调节该轴的激素受体级联反应的效应分子,它的分子结构或功能异常,将影响靶基因IGF-1与其结合,从而引起生长障碍,可能与特发性矮小(ISS)发生有一定关系.国内外尚无IGF-1R基因与ISS的研究,为探讨其与ISS的关系,现就近年来有关IGF-1 R与人体生长障碍的研究作简要综述.     

胰岛素样生长因子-1在新生儿高胆红素血症中的变化及意义

目的：胰岛素样生长因子-1(IGF-1)是神经系统必需的调节因子,目前少有报道其与高胆红素血症之间的关系。该文主要通过测定高胆红素血症（高胆）新生儿血清中IGF-1水平及新生儿神经行为评分(NBNA)来探讨IGF-1与高胆的关系及其临床意义。方法：应用电化学发光分析法检测57例高胆新生儿和 25例正常新生儿血清中IGF-1 浓度,同步测定血清总胆红素(TSB)、未结合胆红素(USB)及白蛋白(ALB)含量,计算USB与ALB比值(B/A),并行新生儿 NBNA 评分。高胆组按血清TSB值221~256 μmol/L,257~342 μmol/L,＞342 μmol/L分为轻、中、重三组;对照组TSB ＜85 μmol/L。结果：轻、中、重高胆患儿血清IGF-1浓度均值分别为39.38±8.42,30.77±4.65,26.34±2.05 ng/L,较对照组50.16±15.73 ng/ L明显降低,在轻、中、重高胆组间IGF-1浓度差异存在显著性(P＜0.01),其值随着胆红素的升高而降低;轻、中、重高胆组NBNA评分均值分别为35.01±2.26,32.45±2.74,26.77±5.02,明显低于对照组38.24±0.78(P＜0.01),高胆各组间差异也有显著性(P＜0.01);血清IGF-1 浓度与NBNA评分呈正相关(r=0.603, P＜0.01),与B/A值呈负相关(r=-0.483, P＜0.01)。结论：高胆患儿血清IGF-1浓度显著降低,降低程度与血清胆红素水平有关;IGF-1可能与新生儿胆红素脑损伤密切相关。［中国当代儿科杂志,2009,11（5）：357-360］     

HSP70在缺血再灌注大鼠肾脏中的表达及意义

目的观察大鼠肾脏缺血及缺血再灌注(I/R)诱导热休克蛋白70(HSP70)的表达及意义。方法制作大鼠肾脏缺血及I/R模型,缺血组于缺血5、15和30min,I/R组于再灌注2、5、8、24、48、72h不同时间摘取肾脏,采用免疫组化方法检测肾脏HSP70的表达,同时采用HE染色,观察肾脏病理改变。结果在缺血组各时间点肾脏HSP70的表达无明显差异(P>0·05);肾脏I/R后,随时间延长,HSP70表达明显增强(P<0·05);肾脏病理显示,缺血时肾组织呈现局灶性及弥散性血管周围水肿,I/R后随时间延长肾脏损伤加重,损伤部位主要在肾小管,表现为空泡样变、小管萎缩、上皮细胞脱落及肾小球囊腔扩张等。结论HSP70在I/R后表达增强,但肾脏仍损伤严重,提示HSP70在肾脏I/R过程中没有起到主要的保护作用。     

孤独症谱系障碍儿童的血清胰岛素样生长因子-1和胰岛素样生长因子结合蛋白-3水平研究北大核心CSCD

目的探讨孤独症谱系障碍(autism spectrum disorder,ASD)儿童的血清胰岛素样生长因子-1(insulin-like growth factor-1,IGF-1)和胰岛素样生长因子结合蛋白-3(insulin-like growth factor binding protein-3,IGFBP-3)水平及与孤独症核心症状之间的关系。方法前瞻性选取重庆市妇幼保健院门诊招募的150名2~7岁ASD儿童和165名年龄、性别相匹配的正常健康儿童为研究对象,采用孤独症行为量表和孤独症评定量表评估ASD儿童核心症状,采用化学发光法检测两组儿童血清IGF-1和IGFBP-3水平。结果ASD组儿童血清IGF-1水平低于对照组儿童(P<0.05)。重度ASD儿童血清IGF-1和IGFBP-3水平低于轻-中度ASD儿童(P<0.001),2~3岁ASD儿童血清IGF-1水平低于对照组儿童(P<0.05)。两组男童IGF-1水平均低于女童(P<0.05)。血清IGF-1、IGFBP-3水平与儿童孤独症评定量表总分呈负相关(分别r=-0.32、-0.40,均P<0.001)。结论儿童早期血清IGF-1降低可能与ASD疾病发展相关,血清IGF-1和IGFBP-3水平与ASD儿童核心症状具有一定关联。     

Insulin-like growth factor-1 as an indicator of nutrition during treatment of adolescent girls with eating disorders

AIM: The use of serum insulin-like growth factor-1 (IGF-1) concentrations as an index of nutrition has been analysed in teenage girls with eating disorders and weight loss. METHOD: Blood samples for analysis of IGF-1 were obtained at 349 assessments of 302 patients and biweekly during 56 treatment periods in 46 patients. IGF-1 was related to body size, weight loss, degree of leanness (BMI standard deviation score) and rate of weight loss. RESULTS: At assessment, when most of the girls were on a weight-losing course, serum IGF-1 concentrations were low. Weight loss immediately prior to assessment was the most important predictor of IGF-1. Together with measurements of weight, height, weight loss and BMI standard deviation score the rate of weight loss predicted IGF-1 to 32-55%. During treatment when there was net weight gain, IGF-1 increased in parallel with the BMI standard deviation score, a measure of leanness, and was also influenced by the short-term weight trend. CONCLUSION: The serum IGF-1 concentration is an indicator of nutritional status in adolescents with eating disorders. It is sensitive to short-term weight changes measured in a clinical setting and could be used at assessment and to monitor nutritional rehabilitation.