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.     

mRNA expression of chemokines in rat kidneys with ureteral obstruction

BACKGROUNDS: It has been demonstrated that leukocyte infiltration, mainly of macrophages and lymphocytes, into obstructed kidneys (OBK) of rats during unilateral ureteral obstruction (UUO). Chemokines (C-C subfamily) may be involved in this mechanisms. Thus, we accessed the gene expression of chemokines in renal cortex of rats with UUO. MATERIALS AND METHODS: Female SD rats were sacrificed at various time points after UUO. mRNA expression of MCP-1, RANTES and MIP-1 alpha was determined by semi-quantitative RT-PCR. RESULTS: Control kidneys (CNK) showed a weak mRNA expression of MCP-1, RANTES and MIP-1 alpha. OBKs showed an increase in MCP-1 at 2 hours of UUO and a significant increase at 4 hours of UUO as compared with CNKs or contralateral unobstructed kidneys (CLK). The mRNA levels of RANTES and MIP-1 alpha were not increased until 72 hours of UUO in CLKs or OBKs. There were slight, but significant, differences of RANTES and MIP-1 alpha expression between OBKs and CNKs at 120 hours of UUO. CONCLUSIONS: We suggest that the early increase in MCP-1 contributes to the leukocyte infiltration and that RANTES and MIP-1 alpha plays a partial role in a late increases.     

血管内皮生长因子及其受体在单侧输尿管梗阻大鼠术侧肾脏的表达

血管内皮生长因子（VEGF）的主要作用是刺激血管内皮细胞的增殖与分化、抑制内皮细胞凋亡，主要通过两个受体Flk-1和Flt-1发挥作用。本研究旨在明确VEGF及其受体在单侧输尿管梗阻（UUO）大鼠术侧肾脏不同时间的表达变化情况，为进一步探讨其在UUO大鼠小管间质纤维化中的作用机制提供实验依据。     

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。     

Effects of acute ureteral obstruction on glomerular hemodynamics in rat kidney

In order to study the effects of acute ureteral obstruction on glomerular hemodynamics, glomerular hydrostatic capillary pressure (PG), pressure in the first-order peritubular capillaries (EAP), and intratubular pressure (PT) were directly measured in superficial nephrons on Munich-Wistar rats by micropuncture with a servo-nulling device, in control conditions and one to two hours after ureteral ligation. Single nephron filtration fraction (SNFF) was calculated from arterial and peritubular blood protein concentration. SNGFR was measured by conventional micropuncture techniques in control conditions and was calculated from efferent arteriole blood flow (EABF) and SNFF during ureteral obstruction. EABF was obtained by timed complete collection of blood from superficial efferent arterioles. Afferent arteriole blood flow (AABF) and resistance of afferent (Ra) and efferent arterioles (Re) were calculated from conventional equations. Ureteral obstruction markedly increased PT from 12.9 +/- 1.4 to 36.8 +/- 6.1 (SD) mm Hg. The fall in SNGFR (from 23.3 +/- 6.4 to 17.9 +/- 5.2 [SD] nl/min) was blunted by the rise in PG (from 45.5 +/- 3.6 to 59.3 +/- 4.0 [SD] mm Hg) and AABF (from 130.0 +/- 59.1 to 144.2 +/- 69.0 [SD] nl/min), secondary to a fall in Ra. These results demonstrate that SNGFR is maintained early after complete ureteral obstruction because of afferent arteriole dilatation.     

Effect of ureteral obstruction on proximal tubular functions of rat kidney

The effect of complete unilateral ureteral obstruction (CUUO) on proximal tubular functions was studied in rats, using the K+-sensitive microelectrode technique and split oil drop method. In the control kidneys peritubular membrane potential (EMperi) and intracellular potassium activity (aKi) were -70.8 +/- 7.0 mV and 81.2 +/- 22.0 mEq/liter (mean +/- SD), respectively. In the CUUO kidneys both EMperi and aKi were progressively reduced with the duration of obstruction. However, in all tubules aKi values were still above the electrochemical equilibrium. In the three days' CUUO kidneys EMperi and aKi were -51.5 +/- 11.7 mV and 53.8 +/- 22.8 mEq/liter, respectively. Rate of fluid absorption (JVL;nl./sec. mm.) across the proximal tubular epithelium from Ringer solution in the control and three days' CUUO kidneys was 0.029 and 0.0065 respectively. In the CUUO kidneys there were wide variations in JVL and EMperi, but there was a clear correlation between these two variables. JVL from choline chloride solution was negligible in both control and CUUO kidneys. From above results, it was suggested that the proximal tubular reabsorption primarily depending on the Na+-K+ pump might be reduced but still working in the CUUO kidney, and thus the proximal tubular reabsorption might take part in preservation of glomerular filtration during the obstruction.     

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。     

Long-term partial ureteral obstruction and its effects on kidney function.

Previously it has been shown that partial ureteral obstruction present in young rats for 12 weeks results in small morphological changes in the kidney as well as slightly decreased kidney function. In the present study the aim was to examine whether rats obstructed for one year had more advanced changes in morphology and kidney function. The first group of animals examined after three weeks of obstruction showed only modest changes in kidney function with a reduced potassium concentration in the urine but no reduction in the glomerular filtration rate. After one year there was a reduction in urine flow as well as in the excretion of both potassium and sodium. Urine osmolality was also reduced. Glomerular filtration rate measured in this group of animals was reduced in the obstructed kidney by about 60% compared to the contralateral one. There were only small changes in the morphology with no loss in parenchymal weight or compensatory hypertrophy, but there was a significant deformation of the papilla and an increase in inflammatory cells in the parenchyma. In conclusion hydronephrosis during a shorter period is not harmful to kidney function but if sustained for an extended time period kidney function will deteriorate.     

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。     

Osteopontin及其mRNA在正常大鼠肾脏的表达

为了研究Ｏｓｔｅｏｐｏｎｔｉｎ（ＯＰ）及其ｍＲＮＡ在正常大鼠肾脏的表达，分别采用抗ＯＰ的单克隆和多克隆抗体进行免疫组化、用地高辛标记的ＯＰｃＲＮＡ探针进行原位杂交。结果表明：ＯＰ及其ｍＲＮＡ在大鼠肾脏远曲小管、集合管呈阳性染色。由于其表达部位是尿中形成结石晶体各种矿物质的高浓缩区，作者认为：Ｏｓｔｅｏｐｏｎｔｉｎ可能为正常机体一种重要的对结石形成产生抑制的内源性大分子物质。     

Prospective evaluation of ureteral stent durability in patients with chronic ureteral obstruction

Objectives. To prospectively evaluate the durability of ureteral stents in patients with chronic ureteral obstruction.Methods. Twenty-nine renal units from 22 patients with known chronic ureteral obstruction treated with indwelling ureteral stents were studied at 3, 6, 9, and 12 months after initial stent placement (7 to 8F Percuflex) by abdominal x-ray, serum creatinine, and renal and bladder ultrasonography. Renal ultrasound scans were performed by a single senior radiologist with specific attention paid to any increase in hydronephrosis and to the presence of ureteral jets. The stent was changed when calcifications were seen along the stent on abdominal x-ray, if an increase in serum creatinine occurred, or if any increase in hydronephrosis or absence of ureteral jets was seen on ultrasonography. In addition, the stents were changed if new-onset flank pain and/or pyelonephritis developed. If none of the above occurred, the stents were changed after 12 months.Results. Two patients died of metastatic cancer, leaving 25 renal units available for follow-up. The mean and median time to stent change were 6 months. At 3, 6, 9, and 12 months, the number of renal units that did not meet the criteria warranting stent change was 18 (72%), 12 (48%), 6 (24%), and 2 (8%), respectively. Only 2 (8%) of 25 renal units did not meet any exclusion criteria at 12 months. The most common reasons for requiring the stent to be changed were new-onset flank pain in 8 (35%), an increase in serum creatinine in 7 (30%), and new-onset or increased hydronephrosis in 7 (30%). No criteria were useful in predicting how long stents could be maintained in an individual patient.Conclusions. The stents were maintained for more than 6 months in 12 (48%) of 25 renal units. However, very few patients were able to maintain chronic ureteral stents for 12 months without requiring stent change based on our criteria.     

Effect of 24-hour ureteral obstruction on subsequent ischemic damage in rat kidney

The effects of prior 24-hour ureteral obstruction on ischemic renal damage were studied in rats. Rats were divided into 6 groups with different times of ischemia (0, 60 and 90 min) and with or without 24-hour ureteral obstruction. Following a 4-week recovery period, contralateral nephrectomy was performed and the rat was sacrificed 24 h later for the determination of serum creatinine and for histologic examination of the affected kidney. A preceding ureteral obstruction for 24 h made no difference to the renal damage with 60 min of ischemia or without ischemia. However, kidneys with 90 min of ischemia and 24 h of ureteral obstruction were more damaged than those with 90 min of ischemia only. These results suggested that the hydronephrotic kidney was more susceptible to long periods of ischemia than the normal kidney.     

Variable chronic partial ureteral obstruction in the neonatal rat: a new model of ureteropelvic junction obstruction

BACKGROUND: Congenital ureteropelvic junction (UPJ) obstruction is a common developmental anomaly. To elucidate the mechanisms underlying the renal consequences of congenital UPJ obstruction, we have developed a new model of variable partial unilateral ureteral obstruction (UUO) in the neonatal rat. METHODS: Rat pups were subjected to sham-operation, complete UUO, or variable partial UUO within the first day of life. After 14 or 28 days, the relative number of glomeruli, cell proliferation, tubular apoptosis, tubular atrophy, and interstitial fibrosis were quantitated in histologic sections. Glomerular filtration rate (GFR) was determined after 28 days of partial or complete UUO. RESULTS: Following 70% to 75% reduction in ureteral diameter, renal growth from 14 to 28 days was reduced by 60%, and the number of glomeruli decreased by 50%. Renal pelvic diameter increased in proportion to the severity of obstruction following 14 days of partial UUO, and by 28 days, was maximally dilated regardless of the luminal diameter. Renal proliferation was increased, while tubular apoptosis, tubular atrophy, and interstitial fibrosis were less severe 14 days following partial UUO than in complete UUO. GFR was reduced by 80%, and proteinuria developed following 28 days of partial UUO. CONCLUSION: Renal function is impaired by chronic ipsilateral partial UUO, which reduces the number of nephrons, and leads to progressive renal pelvic dilatation. Tubular atrophy and interstitial fibrosis develop prior to significant renal pelvic dilatation. Correlation of clinically measurable parameters with renal morphometry or imaging studies in this model may lead to new approaches to the management of congenital UPJ obstruction.     

烧伤后肾组织内皮素-1 及其受体基因的表达

目的探讨烧伤后肾组织内皮素－１（ＥＴ－１）升高的效应部位以及效应的ＥＴＲ亚型。方法采用斑点杂交和原位杂交技术对大鼠３０％体表面积Ⅲ度烧伤模型的ＥＴ－１及其受体亚型（ＥＴＡ、ＥＴＢ）的基因表达进行了研究。结果烧伤后１小时，ＥＴ－１、ＥＴＡ、ＥＴＢ、ｍＲＮＡ均明显升高；烧伤后２４小时仍维持较高水平。ＥＴ－１和ＥＴＡｍＲＮＡ杂交信号于烧伤后３小时达高峰。ＥＴ－１ｍＲＮＡ主要分布肾皮质小血管内皮细胞、髓质肾小管和集合管，ＥＴＡ受体ｍＲＮＡ则分布于上述小血管的平滑肌细胞。ＥＴＢ受体ｍＲＮＡ于烧伤后６小时达高峰，主要分布髓质肾小管、集合管。结论烧伤后肾脏内皮素受体亚型上调在皮质以ＥＴＡ为主，在髓质以ＥＴＢ为主，它们分别与增强表达的ＥＴ－１结合可能导致肾皮质缺血和水钠代谢异常，是烧伤后肾功能障碍的重要因素。     

Expression of transforming growth factor-β type II receptor mRNA in embryonic and adult rat kidney

Summary: The transforming growth factor-β (TGF-β) family of growth factors regulates cell proliferation, differentiation, extracellular matrix synthesis and angiogenesis in many developing tissues. Transforming growth factor-β1 was recently shown to affect the branching of ureteric epithelium and nephron formation in cultured rat metanephroi. As the TGF-β type II receptor is specific for the TGF-β family, the present study used in situ hybridization to localize mRNA for this receptor in metanephroi from Sprague-Dawley rat embryos. Transforming growth factor-β type II receptor mRNA was located in ureteric duct epithelium, undifferentiated mesenchymal cells in the nephrogenic zone, vesicles, comma-shaped bodies and S-shaped bodies. In some S-shaped bodies, TGF-β type II receptor mRNA was not expressed in the lower limb, which subsequently forms the renal corpuscle. Expression was not observed in capillary loop stage glomeruli and maturing glomeruli, or in proximal tubules and interstitial cells. In adult rat kidney, TGF-β type II receptor mRNA was expressed in cortical collecting ducts and distal tubules but not in glomeruli or proximal tubules. These findings demonstrate that the prominent expression of TGF-β type II receptor mRNA decreases as glomeruli and tubules develop. Expression then remains undetectable in adult glomeruli and proximal tubules. the developmentally-regulated expression of this receptor suggests a key role in glomerular and nephron development.     

烧伤后肾组织内皮素-1及其受体基因的表达

目的探讨烧伤后肾组织内皮素-1(ET-1)升高的效应部位以及效应的 ETR 亚型。方法采用斑点杂交和原位杂交技术对大鼠30％体表面积Ⅲ度烧伤模型的 ET-1及其受体亚型(ETA、ETB)的基因表达进行了研究。结果烧伤后1小时,ET-1、ETA、ETB、mRNA 均明显升高;烧伤后24小时仍维持较高水平。ET-1和 ETA mRNA 杂交信号于烧伤后3小时达高峰。ET-1mRNA 主要分布肾皮质小血管内皮细胞、髓质肾小管和集合管,ETA 受体 mRNA 则分布于上述小血管的平滑肌细胞。ETB 受体mRNA 于烧伤后6小时达高峰,主要分布髓质肾小管、集合管。结论烧伤后肾脏内皮素受体亚型上调在皮质以 ETA 为主,在髓质以 ETB 为主,它们分别与增强表达的 ET-1结合可能导致肾皮质缺血和水钠代谢异常,是烧伤后肾功能障碍的重要因素。     

缺血再灌流肾组织内皮素—1及其受体基因表达的实验研究

为了探究内皮素１（ＥＴ１）对肾功能的影响和作用方式，采用斑点杂交和原位杂交方法对大鼠缺血６０分钟再灌注肾组织ＥＴ１及其受体亚型（ＥＴＡ、ＥＴＢ）的基因表达进行了研究。结果发现：再灌流１小时，ＥＴ１、ＥＴＡ、ＥＴＢｍＲＮＡ均明显升高；再灌流２４小时仍维持较高水平。ＥＴ１和ＥＴＡｍＲＮＡ杂交信号再灌流３小时达高峰。ＥＴ１ｍＲＮＡ主要分布肾皮质小血管内皮细胞、髓质肾小管和集合管，ＥＴＡ受体ｍＲＮＡ则分布于上述小血管的平滑肌细胞。ＥＴＢ受体ｍＲＮＡ于再灌流６小时达高峰，主要分布髓质肾小管、集合管。说明缺血再灌流肾内皮素受体亚型上调在皮质以ＥＴＡ为主，在髓质以ＥＴＢ为主，分别与增强表达的ＥＴ１结合导致肾皮质缺血和水钠代谢异常。