新型免疫调节剂FTY720对肾间质纤维化的防治及机制探讨
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摘要
目的观察单侧输尿管梗阻(unilateral ureteral obstruction,UUO)诱导的大鼠肾间质纤维化模型中,肾间质纤维化病变及管周毛细血管的丢失情况,同时用新型免疫调节剂FTY720对其进行干预,观察其对肾间质纤维化程度及管周毛细血管丢失的影响及防治作用,并探讨其作用机制。
方法将45只SD大鼠随机分为三组,即假手术组(SHAM),单侧输尿管梗阻模型组(UUO)和FTY720治疗组(FTY720),每组15只。模型组和FTY720治疗组大鼠行单侧输尿管梗阻术,假手术组开腹后不结扎输尿管,缝合腹腔,作为对照组。治疗组于术前三天给予FTY720 0.5mg·kg-1·d-1灌胃,模型组和假手术组给予等量生理盐水,三组灌胃直至处死动物。每组分别于术后第7﹑14﹑21天各处死大鼠五只,每组大鼠分别于处死前放入代谢笼留取24小时尿液,进行尿蛋白定量。并通过心脏采血,取血清测定血Scr、BUN。处死大鼠后留取梗阻侧肾组织做HE和MASSON染色观察肾间质纤维化程度,同时用Tunel法检测肾小管细胞凋亡情况,用免疫组化方法测定肾组织中CD3、ED1、α-平滑肌肌动蛋白(α-SMA)、胶原-Ⅲ(COL-Ⅲ)、CD141、VEGF的表达水平。
结果24h尿蛋白定量,术后14d模型组为(15.48±1.52 )mg·d-1, FTY720治疗组为(9.7±1.74)mg·d-1,较模型组明显减低,差异有统计学意义(P<0.05)。术后7、14、21天,模型组血肌酐(Scr)水平分别为(123±15.76)μmol·L-1、(65.88±6.08)μmol·L-1、(51.38±6.9)μmol·L-1,FTY720治疗组血肌酐相应为(41.76±4.51)μmol·L-1、(39.54±7.94)μmol·L-1、(38.5±4.52)μmol·L-1,均明显降低(P<0.05),FTY720治疗组与假手术组比较差异无统计学意义。术后7、14天,模型组BUN分别为(16.14±3.21)mmol·L-1和(10.7±1.66)mmol·L-1,FTY720治疗组BUN分别为(8.08±0.96 )mmol·L-1和(6.98±0.80)mmol·L-1,均有一定程度下降,差异有统计学意义(P<0.05)。病理学检查显示,FTY720治疗组肾间质纤维化程度较模型组明显减轻。免疫组化结果显示,假手术组肾间质各时间点均未见炎性细胞浸润,模型组术后第7天肾间质出现大量CD3+、ED1+细胞浸润,第14、21天明显增多,与模型相比,FTY720治疗组肾间质CD3+、ED1+细胞数有一定程度地降低(P<0.01)。α-SMA表达在假手术组中仅限于血管,而在FTY720治疗组和UUO模型组中,表达明显增多,可见于肾小管和间质细胞,但FTY720治疗组中表达要弱于模型组。假手术组中各个时间点肾间质中可见少量Ⅲ型胶原表达,模型组手术后7、14和21dⅢ型胶原表达明显增强,主要集中于肾小管间质病变明显区域,FTY720治疗组中表达则明显减少。假手术组肾间质微血管未见明显病变,术后7、14、21天CD141阳性毛细血管密度分别为158.6±8.29、155.2±5.89、157±4.47,模型组肾间质微血管病变严重,主要集中于肾间质小管损伤明显区域, CD141阳性毛细血管数进行性减少,术后7、14、21天分别为116.8±8.92、98.8±3.96、62.2±7.01。FTY720治疗可明显减轻毛细血管病变及丢失,术后7、14、21天毛细血管密度分别为132.5±8.32、119.4±12.21、85±2.59,均显著高于模型组(P<0.01)。免疫组化结果显示,假手术组各时间点均未见明显凋亡,模型组术后第7天已见肾小管上皮细胞凋亡,随梗阻时间延长,凋亡细胞呈明显增多趋势,而FTY720治疗组各时间段肾小管上皮细胞凋亡数均低于模型组(P<0.01)。假手术组肾小管上皮细胞可见较多VEFG蛋白表达,模型组术后第7、14、21天VEFG蛋白表达量呈逐渐降低趋势,炎性细胞浸润明显处几乎无VEGF表达。与模型组相比,FTY720治疗组肾小管上皮细胞VEGF蛋白表达量高于模型组(P<0.01),但低于假手术组(P<0.05)。
结论新型免疫调节剂FTY720能明显减少UUO大鼠肾间质的T淋巴细胞和巨噬细胞的浸润,从而减轻肾小管上皮细胞调亡,上调VEGF表达,减少肾间质毛细血管丢失,并且FTY720在一定程度上可以抑制肾小管上皮细胞转分化和细胞外基质的积聚,明显减轻单侧输尿管梗阻后大鼠肾间质纤维化的程度,对肾功能有一定的保护作用。
【Objective】FTY720 is a novel immune modulating drug which obtained through structural modification of ISP-1, an immunosuppressive substance newly purified from Cordyceps sinensis , a Chinese herb traditionally used in the treatment of renal diseases. In recent years, the role of FTY720 in the immune mediated glomerulonephritis has been demonstrated, but its effects on renal interstitial fibrosis, a major determinant for prognosis, remain uncertain.. The present study is aimed to examine the inhibitory effect of FTY720 on renal interstitial fibrosis and peritubular cappilary loss in unilateral ureteral obstructic rats and to study the underlying mechanism .
【Methods】Fouty-five males SD rats were randomly divided into sham-operated(SHAM) , unilateral ureteral obstruction (UUO) and UUO treated with FTY720 (UUO+FTY720) group . 0.5mg.kg-1.d-1 of FTY720 or vehicle was administrated through daily gavage and begun from three days before the operation till being sacrificed. 5 rats in each groups were sacrificed at day 7, day 14 and day 21 afte UUO or Sham-operation. In order to document if renal fibrosis closely correlates with renal insufficiency, 24-hour urine protein, blood urea nitrogen (BUN) and plasma creatinine were determined .Tubulointerstitial injury and interstitial fibrosis were examined by morphological analysis, apoptosis of tubular epithelial cells was examined by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling , the expression of CD3,ED1 ,α-SMA ,COL-Ⅲ,CD141and VEGF was detected by immunohistochemical nethod and scored semi-quantitively. The number of CD141 positive PTC was quantitated in an area of 100 renal tubules.
【Results】The amount of 24 hours urine protein was ((9.7±1.74)mg·d-1 at 14d after operation in FTY720 treated group, much lower than that in UUO group, which was (15.48±1.52 )mg·d-1 at same time .(P<0.05). Serum creatinine were (123±15.76)μmol·L-1,(65.88±6.08)μmol·L-1(,51.38±6.9)μmol·L-1 in UUO groups respectively at 7,14,21d, and (41.76±4.51)μmol·L-1,(39.54±7.94)μmol·L-1,(38.5±4.52)μmol·L-1 in FTY720 treated group at 7,14,21d, the levels in FTY720 treated group were significantly lower than those in UUO group. The Blood urea nitrogen was (16.14±3.21)mmol·L-1(,10.7±1.66)mmol·L-1 in UUO group and (8.08±0.96 )mmol·L-1(,6.98±0.80)mmol·L-1 in FTY720 treated group at 7,14d respectively, the levels were significantly lower in FTY720 group than that in UUO group (P<0.05). The renal pathological evaluation indicated that the scores of renal interstitial fibrosis were lower in FTY720 group than that in UUO group. Obvious lymphocyte and macrophage infiltration were found in tubular interstitial area in UUO group but significantly less in FTY720 treated group (P<0.01). Immunohistochemistry study revealed thatα-SMA expression was limited to vessels in SHAM group, but extended to renal tubule and interstitium in UUO and FTY720 treated group, while relatively weaker expression was observed in FTY720 group than in UUO group . Some collagenⅢexpression was found in SHAM group, which was much enhanced in UUO group at 7,14,21d and mainly distributed in renal interstitium, the expression in FTY720 group was also increased compared to SHAM group, but much lower than that in UUO group. In the SHAM group , there were no significant injury in peritubular capillary, and the numbers of CD141 positive PTC were 158.6±8.29, 155.2±5.89, 157±4.47 at 7, 14, 21 day. In the UUO group, peritubular capillary injuried severely ,and peritubular capillary density decreased markedly, the numbers of CD141 positive PTC were 116.8±8.92, 98.8±3.96, 62.2±7.01 at the 7th,14th,21st day. FTY720 could relieve the impairment and loss of peritubular capillary, as demonstrated in FTY720 treated group,the numbers of CD141 positive PTC were 132.5±8.32, 119.4±12.21, 85±2.59 at the 7th,14th,21st day ,significantly higher than those in UUO group(P<0.01)., No tubular epithelial cells apoptosis was observed in SHAM group, apoptosis was found in UUO group at day 7 after operation but was decreased in FTY720 treated group (P<0.01). The immunohistochemistry study revealed that the expression of VEGF protein was higher in FTY720 treated group than in UUO group(P<0.01),but still lower than in SHAM group(P<0.05).
【Conclusions】Novel immunomodulator FTY720 can obviously inhibite lymphocyte and macrophage infiltration, renal tubule cell transdifferentiation, extracellular matrix accumulation and inhibit the tubular epithelial cell apoptosis, increase the expression of VEGF protein ,and prevent the loss of peritubular capillary interstitial fibrosis, thus ameliorate renal interstitial fibrosis and prevent renal disease progression.
方法将45只SD大鼠随机分为三组,即假手术组(SHAM),单侧输尿管梗阻模型组(UUO)和FTY720治疗组(FTY720),每组15只。模型组和FTY720治疗组大鼠行单侧输尿管梗阻术,假手术组开腹后不结扎输尿管,缝合腹腔,作为对照组。治疗组于术前三天给予FTY720 0.5mg·kg-1·d-1灌胃,模型组和假手术组给予等量生理盐水,三组灌胃直至处死动物。每组分别于术后第7﹑14﹑21天各处死大鼠五只,每组大鼠分别于处死前放入代谢笼留取24小时尿液,进行尿蛋白定量。并通过心脏采血,取血清测定血Scr、BUN。处死大鼠后留取梗阻侧肾组织做HE和MASSON染色观察肾间质纤维化程度,同时用Tunel法检测肾小管细胞凋亡情况,用免疫组化方法测定肾组织中CD3、ED1、α-平滑肌肌动蛋白(α-SMA)、胶原-Ⅲ(COL-Ⅲ)、CD141、VEGF的表达水平。
结果24h尿蛋白定量,术后14d模型组为(15.48±1.52 )mg·d-1, FTY720治疗组为(9.7±1.74)mg·d-1,较模型组明显减低,差异有统计学意义(P<0.05)。术后7、14、21天,模型组血肌酐(Scr)水平分别为(123±15.76)μmol·L-1、(65.88±6.08)μmol·L-1、(51.38±6.9)μmol·L-1,FTY720治疗组血肌酐相应为(41.76±4.51)μmol·L-1、(39.54±7.94)μmol·L-1、(38.5±4.52)μmol·L-1,均明显降低(P<0.05),FTY720治疗组与假手术组比较差异无统计学意义。术后7、14天,模型组BUN分别为(16.14±3.21)mmol·L-1和(10.7±1.66)mmol·L-1,FTY720治疗组BUN分别为(8.08±0.96 )mmol·L-1和(6.98±0.80)mmol·L-1,均有一定程度下降,差异有统计学意义(P<0.05)。病理学检查显示,FTY720治疗组肾间质纤维化程度较模型组明显减轻。免疫组化结果显示,假手术组肾间质各时间点均未见炎性细胞浸润,模型组术后第7天肾间质出现大量CD3+、ED1+细胞浸润,第14、21天明显增多,与模型相比,FTY720治疗组肾间质CD3+、ED1+细胞数有一定程度地降低(P<0.01)。α-SMA表达在假手术组中仅限于血管,而在FTY720治疗组和UUO模型组中,表达明显增多,可见于肾小管和间质细胞,但FTY720治疗组中表达要弱于模型组。假手术组中各个时间点肾间质中可见少量Ⅲ型胶原表达,模型组手术后7、14和21dⅢ型胶原表达明显增强,主要集中于肾小管间质病变明显区域,FTY720治疗组中表达则明显减少。假手术组肾间质微血管未见明显病变,术后7、14、21天CD141阳性毛细血管密度分别为158.6±8.29、155.2±5.89、157±4.47,模型组肾间质微血管病变严重,主要集中于肾间质小管损伤明显区域, CD141阳性毛细血管数进行性减少,术后7、14、21天分别为116.8±8.92、98.8±3.96、62.2±7.01。FTY720治疗可明显减轻毛细血管病变及丢失,术后7、14、21天毛细血管密度分别为132.5±8.32、119.4±12.21、85±2.59,均显著高于模型组(P<0.01)。免疫组化结果显示,假手术组各时间点均未见明显凋亡,模型组术后第7天已见肾小管上皮细胞凋亡,随梗阻时间延长,凋亡细胞呈明显增多趋势,而FTY720治疗组各时间段肾小管上皮细胞凋亡数均低于模型组(P<0.01)。假手术组肾小管上皮细胞可见较多VEFG蛋白表达,模型组术后第7、14、21天VEFG蛋白表达量呈逐渐降低趋势,炎性细胞浸润明显处几乎无VEGF表达。与模型组相比,FTY720治疗组肾小管上皮细胞VEGF蛋白表达量高于模型组(P<0.01),但低于假手术组(P<0.05)。
结论新型免疫调节剂FTY720能明显减少UUO大鼠肾间质的T淋巴细胞和巨噬细胞的浸润,从而减轻肾小管上皮细胞调亡,上调VEGF表达,减少肾间质毛细血管丢失,并且FTY720在一定程度上可以抑制肾小管上皮细胞转分化和细胞外基质的积聚,明显减轻单侧输尿管梗阻后大鼠肾间质纤维化的程度,对肾功能有一定的保护作用。
【Objective】FTY720 is a novel immune modulating drug which obtained through structural modification of ISP-1, an immunosuppressive substance newly purified from Cordyceps sinensis , a Chinese herb traditionally used in the treatment of renal diseases. In recent years, the role of FTY720 in the immune mediated glomerulonephritis has been demonstrated, but its effects on renal interstitial fibrosis, a major determinant for prognosis, remain uncertain.. The present study is aimed to examine the inhibitory effect of FTY720 on renal interstitial fibrosis and peritubular cappilary loss in unilateral ureteral obstructic rats and to study the underlying mechanism .
【Methods】Fouty-five males SD rats were randomly divided into sham-operated(SHAM) , unilateral ureteral obstruction (UUO) and UUO treated with FTY720 (UUO+FTY720) group . 0.5mg.kg-1.d-1 of FTY720 or vehicle was administrated through daily gavage and begun from three days before the operation till being sacrificed. 5 rats in each groups were sacrificed at day 7, day 14 and day 21 afte UUO or Sham-operation. In order to document if renal fibrosis closely correlates with renal insufficiency, 24-hour urine protein, blood urea nitrogen (BUN) and plasma creatinine were determined .Tubulointerstitial injury and interstitial fibrosis were examined by morphological analysis, apoptosis of tubular epithelial cells was examined by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling , the expression of CD3,ED1 ,α-SMA ,COL-Ⅲ,CD141and VEGF was detected by immunohistochemical nethod and scored semi-quantitively. The number of CD141 positive PTC was quantitated in an area of 100 renal tubules.
【Results】The amount of 24 hours urine protein was ((9.7±1.74)mg·d-1 at 14d after operation in FTY720 treated group, much lower than that in UUO group, which was (15.48±1.52 )mg·d-1 at same time .(P<0.05). Serum creatinine were (123±15.76)μmol·L-1,(65.88±6.08)μmol·L-1(,51.38±6.9)μmol·L-1 in UUO groups respectively at 7,14,21d, and (41.76±4.51)μmol·L-1,(39.54±7.94)μmol·L-1,(38.5±4.52)μmol·L-1 in FTY720 treated group at 7,14,21d, the levels in FTY720 treated group were significantly lower than those in UUO group. The Blood urea nitrogen was (16.14±3.21)mmol·L-1(,10.7±1.66)mmol·L-1 in UUO group and (8.08±0.96 )mmol·L-1(,6.98±0.80)mmol·L-1 in FTY720 treated group at 7,14d respectively, the levels were significantly lower in FTY720 group than that in UUO group (P<0.05). The renal pathological evaluation indicated that the scores of renal interstitial fibrosis were lower in FTY720 group than that in UUO group. Obvious lymphocyte and macrophage infiltration were found in tubular interstitial area in UUO group but significantly less in FTY720 treated group (P<0.01). Immunohistochemistry study revealed thatα-SMA expression was limited to vessels in SHAM group, but extended to renal tubule and interstitium in UUO and FTY720 treated group, while relatively weaker expression was observed in FTY720 group than in UUO group . Some collagenⅢexpression was found in SHAM group, which was much enhanced in UUO group at 7,14,21d and mainly distributed in renal interstitium, the expression in FTY720 group was also increased compared to SHAM group, but much lower than that in UUO group. In the SHAM group , there were no significant injury in peritubular capillary, and the numbers of CD141 positive PTC were 158.6±8.29, 155.2±5.89, 157±4.47 at 7, 14, 21 day. In the UUO group, peritubular capillary injuried severely ,and peritubular capillary density decreased markedly, the numbers of CD141 positive PTC were 116.8±8.92, 98.8±3.96, 62.2±7.01 at the 7th,14th,21st day. FTY720 could relieve the impairment and loss of peritubular capillary, as demonstrated in FTY720 treated group,the numbers of CD141 positive PTC were 132.5±8.32, 119.4±12.21, 85±2.59 at the 7th,14th,21st day ,significantly higher than those in UUO group(P<0.01)., No tubular epithelial cells apoptosis was observed in SHAM group, apoptosis was found in UUO group at day 7 after operation but was decreased in FTY720 treated group (P<0.01). The immunohistochemistry study revealed that the expression of VEGF protein was higher in FTY720 treated group than in UUO group(P<0.01),but still lower than in SHAM group(P<0.05).
【Conclusions】Novel immunomodulator FTY720 can obviously inhibite lymphocyte and macrophage infiltration, renal tubule cell transdifferentiation, extracellular matrix accumulation and inhibit the tubular epithelial cell apoptosis, increase the expression of VEGF protein ,and prevent the loss of peritubular capillary interstitial fibrosis, thus ameliorate renal interstitial fibrosis and prevent renal disease progression.
引文
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[23]邓跃毅,陈以平,贺学林,等.冬虫夏草制剂延缓慢性肾衰竭的机理研究.中国中西医结合肾病杂志.2001,2(7):381-383
[24]孙尔维,张雷.一种新型免疫抑制剂-FTY720.国外医学免疫学分册. 2000,23(1):22-25
[25] Klahr S, Morrissey J, et al Obstructive nephropathy and renal fibrosis .Am J Physiol Renal Physiol , 2002, 283(5) : 861-875
[26] Yang , Jun Wei , Liu You Hua . Blockage of tubular epithelial to myofibroblast transition by hepatocyte growth factor prevents renal interstitial fibrosis [J] . J Am Soc Nephrol , 2002 , 13(1): 96-107
[27] Truong . Cell apoptosis and proferation in experimental chronic obstructive uropathy [J]. Kidney Int , 1996 , 50:200-207
[28] Strutz F , Neilson EG . The role of lymphocytes in the progression of interstitial disease . Kidney Int , 1994 , 45(Suppl 45):S106-S110
[29] Postlethwaite AE , Holness MA , Katai H , et al . Human fibroblasts synethesize elevated levels of extracellular matrix proteins in response to interleukin 4. J Clin Invest , 1992,90:1479-1485
[30] Ricardo SD , Diamond JR . The role of macrophages and reactive oxygen species in experimental hydronephrosis . Semin Nephrol , 1998,18:612-621
[31] Suleiman M, Cury PM , Pestana JO ,et al. FTY720 prevents renal T-cell infiltration after ischemia/reperfusion injury . 2005 , 37(1):373-4
[32] Okada H , Strutz F , Danoff TM , et al . Possible pathogenesis of renal fibrosis . Kidney Int , 1996 , 49(Suppl 54):S37-38
[33] Kalluri R , Neilson EG . Epithelial– mesenchymal transition and its implications for fibrosis . J Clin Invest , 2003 , 112:1773-1784
[34] Klahr S , Morrissey J . Obstructive nephropathy and renal fibrosis [J] . Am J Physiol Renal Physiol , 2002, 283(5):861-875
[35] Fine LG , Orphanides C , Normal JT . Progressive renal disease . The chronic hypoxia hypothesis . Kidney Int , 1998 , 65:S74
[36] Fine LG , Orphanides C , Norman JT , et al. Progressive renal disease: the chronichypoxia hypothesis [J] . Kidney Int . 1998 , 65: 74-78
[37] Zhang B , Wang WM , Shi H . Effects of valsartan and ramiprilon on the peritubular microangium in rats with 5/6 subtotal-nephrectomy [J]. J Nephrol Dialy Transplant , 2003 , 12 (4) : 28-31.
[38] Brinkmann V , Pinschewer DD , Feng L , et al . Fty720: altered lymphocyte traffic results in allograft protection . Transplantation , 2001, 72(5) : 764-769
[39] Im DS . Linking Chinese medicine and G-protein-coupled receptors. Trends in Pharamacological Science , 2003, 24(1):2-4
[40] Nagahara Y , Ikekita M , Shinomiya T . Immunosuppressant FTY720 induces apoptosis by direct induction of permeability transition and release of cytochrome from mitochondria [J] . Immunol , 2000 , 165 : 3250-60
[1] Marcussen N , Nyengaard J , Christensen S. Compensatory growth of glomeruli is accomplished by an increased number of glomerular capillaries. Lab Invest , 1994 ,70 :868
[2] Kang DH , Hughes J , Mazzali M, et al.Impaired angiogenesis in the remnant kidney model ( II) : VEGF administration reduces renal fibrosis and stabilizes renal function1 J Am Soc Nephrol ,2001 , 12 :1448
[3] Murohara Y, Horowitz JR , SliverDR , et al.Vascular endothelial growth factor/ vascular permeability factor enhances vascular permeability via nitric oxide and prostacylin1Circulation ,1998 ;97 :99
[4] Thomas S , Vanuystel J , Gruden G, et al. Vascular endothelial growth factor receptors in human mesangium in vitro and in glomerular disease. J Am Soc Nephrol ,2000 , 11 :1236
[5] Yamagishi S , Inagaki Y, Okamoto T , et al. Advanced glycation end product induced apoptosis and overexpression of vascular endothelial growth factor and monocyte chemoattractant protein-1 in human-cultured mesangial cells. J Biol Chem,2002 , 277 :203091
[6] Schweda F , Blumberg FC , Schweda A , et al. Effects of chronic hypoxia on renal PDGF-A , PDGF-B , and VEGF gene expression in rats. Nephron ,2000 ,86 :161
[7] Levy AP. A cellular paradigmfor the failure to increase vascular endothelial growth factor in chronically hypoxic states.Coron Artery Dis , 1999 , 10 :427
[8] Ferrara N . Role of vascular endothelial growth factor in the regulation of angiogenesis. Kidney Int ,1999 ,56 :794
[9] Williams B , Baker A , Gallacher B , et al. Angiotensin II increases vascular permeability factor gene expression by human vascular smooth muscle cells. Hypertension ,1995 ,25 :913
[10] Cha DR , Kim NH , Yoon JW, et al.Role of vascular endothelial growth factor in diabetic nephropathy.Kidney Int ,2000 (Suppl) ,77 :S104
[11] Kang DH , Nakagawa T , Feng L , et al . Nitric oxide modulates vascular disease in the remnant kidney model.AmJ Pathol ,2002 ,161(1) :239
[12] Smith LE , Shen W, Perruzzi C , et al.Regulation of vascular endothelial growth factor-dependent retinal neovascularization by insulin-like growth factor-1 receptor. Nat Med ,1999 ,5 :1390
[13] ZeisbergM, ,Strutz F , Muler GA1 Renal fibrosis : an update .Curr Opin in Nephrol and Hypertens ,2001 ,10 :315
[14] Kang DH , Joly AH , Oh SW, et al.Impaired angiogenesis in the remnant kidney model (Ⅰ) : Potential role of vascular endothelial growth factor and thrombospondin-1. J Am Soc Nephrol ,2001 ,12 :1434
[15] Eremina V , Haigh J , Nagy A , et al. The role of VEGF-A in glomerular angiogenesis [ abstract ].J Am Soc Nephrol ,2002 ,13 :100A
[16] Horita Y, Miyazaki M, Koji T , et al. Expression of vascular endothelial growth factor and its receptors in rats with protein overload nephrosis. Nephrol Dial Transpl ,1998 , 13 :2519
[17] Grone J-J , Simon M, Grone EF. Expression of vascular endothelial growth factor in renal vascular disease and renal allografts. J Pathol , 1995 ,177 :259
[18] Klanke B , Simon M, Rockl W, et al. Effects of vascular endothelial growth factor (VEGF) / vascular permeability factor (VPF) on haemody namics and permselectivity of the isolated perfused rat kidney. Nephrol Dial Transplant ,1998 ,13 :875
[19] Celletti FL , Waugh JM, Amabile PG, et al. Vascular endothelial growth factor enhances atherosclerotic plaque progression. Nat Med , 2001 , 7 :425
[20] Flyvbjerg A , Dagnaes-Hansen F , De Vriese AS , et al . Amelioration of long-term renal changes in obese type 2 diabetic mice by a neutralizing vascular endothelialgrowth factor antibody. Diabetes ,2002 ,51 (10) : 3090-4
[21] El Awad B , Kreft B , Wolber EM , et al . Hypoxia and interleukin-1beta stimulate vascular endothelial growth factor production in human proximal tubular cells . Kidney Int , 2000 Jul ; 58(1) : 43-50
[22] Kanellis J , Frasre S , Katerelos M , et al . Vascular endothelial growth factor is a survival factor for renal tubular epithelial cells .Am J Physiol Renal Physiol , 2000; 278(6): F905-15
[23] Choi YJ , Chakraborty S , Nguyen V , et al . Peritubular capillary loss is associated with chronic tubulointerstitial injury in human kidney : altered expression of vascular endothelial growth factor .Hum Pathol , 2000; 31(12): 1491-7
[24] Cosio F, Alamir A , Yim S , et al . Patient survival after renal transplantation:Ⅰ.The impact of dialysis pre-transplant . Kidney Int , 1998;53(3): 767-72
[25] Fontan M , Rodriguez Carmona A , Garcia Falcon T , et al . Perit Dial Int , 1996; 16:48-51
[26] Ito F , Nakazawa H , Rvoji O , er al . Cytokines accumulated in acquired renal cysts in long term hemodialysis patients. Urol Int . 2000;65(1): 21-7
[27] Ojo AO, Wolfe RA , Held PJ, et al. Delayed graft function : risk factors and implications for renal allograft survival .Transplantation , 1997;63(7):968-74
[28] Kang DH , Kim YG , Andoh TF ,et al . Post-cyclosporine-mediated hypertension and nephropathy: amelioration by vascular endothelial growth factor . Am J Physiol Renal Physiol , 2001 ; 280(4) : F727-36
[29] Pilmore HL , Eris JM , Painter DM , et al . Vascular endothelial growth factor expression in human chronic renal allograft rejection . Transplantation . 1999 ; 67(6) 929-33
[30] Nakamoto Y , Imai H , Yasuda T , et al . A spectrum of clinicpathological features of nephropathy associated with POEMS syndrome . Nephrol Dial Transplant . 1999;14(10):2370-8
[31] Lee JS , Kim HS , Jung JJ , et al . Expression of vascular endothelial growth factor in renal cell carcinoma and the relation to angiogenesis and P53 protein expression . J Surg Oncol .2001; 77(1):55-60
[32] Drevs J , Hofmann I , Hugenschmidt H , et al . Effects of PTK787/ZK 222584 , a specific inhibitor of vascular endothelial growth factor receptor tyrosine kinases , on primary tumor , metastasis , vessel density , and blood flow in a murine renal cell carcinoma model . Cancer Res . 2000;60(17):4819-24
[33] Kuroda N ,Moriki T ,Komatsu F ,et al .Adult-onset giant juxtaglomerular cell tumor of the kidney. Pathol Int ,2000 ;50 (3) : 249～54
[33] Masuda Y, Shimizu A , Mori T , et al. Vascular endothelial growth factor enhances glomerular capillary repair and accelerates resolution of experi2 mentally induced glomerulonephritis1 AmJ Pathol ,2001 ,159 :599
[34]. Kim NH , Oh JH , Seo JA , et al . Vascular endothelial growth factor (VEGF) and soluble VEGF receptor FLT-1 in diabetic nephropathy . Kidney Int . 2005;67(1):166-67
[35] Harada M , Mitsuyama K , Yoshida H , et al . Vascular endothelial growth factor in patients with rheumatoid arthritis.Scand J Rheamatol .1998;27(5):377-80
[2] Eitner , Frank , Floege , Jurgen . Novel insights into renal fibrosis . J Current Opinion in Nephrology and Hypertension , 2003 , 12 (3) : 227 - 232.
[3] Donnell MP . Renal tubulointerstitial fibrosis: new thoughts on its development and progression . Post-grad Med , 2000 , 108(1): 159-172
[4] El Nahas AM , Muchaneta-Kubara EC ,Essawy M , et al . Renal fibrosis : insights into pathogenesis and treatment . Int J Bioche Cell Biol ,1997 , 29(1): 55-62
[5] Norman JT , Clark IM , Garcia PL . Hypoxia promotes fibrogenesis in human renal fibroblasts [J] . Kidney Int , 2000 , 58 (6) : 2351-2366
[6] Choi YJ , Chakraborty S , Nguyen V . Peritubular capillary loss is associated with chronic tubulointerstitial injury in human kidney : altered expression growth factor [J] . Hum Paththol , 2000, 31 (1) : 1491-1497
[7] Fine LG, Orphanides C, Norman J T, et al . Progressive renal disease :the chronic hypoxia hypothesis[J ] . Kidney Int. 1998 ,65(1):74～78
[8] Fujita T , Inoue K , Yamamoto S , et al . Fungal metabolites . A potent immunosuppressive activity found in Isaria sinclairii metabolite . J Immunol , 1994 , 47(2): 208 -215
[9] Chiba K , Yanagawa Y , Masubuchi Y , et al . FTY720 , a novel immunosuppressant , induces sequestration of circulating mature lymphocytes by acceleration of lymphocyte homing in rats . I . FTY720 selectively decreases the number of circulating mature lymphocytes by acceleration of lymphocyte homing . J Immunol , 1998 , 160 : 5037-5044
[10] Peters T , Martini S , Wang Y , et al , Selective lymphocyte inhibition by FTY720 slows the progressive course of chronic anti-thy1 glomerulosclerosis . Kidney Int , 2004 , 66 (4) : 1434-1443
[11]肖朝华,周建华,吴衡生.多球壳菌素对高糖诱导肾小球系膜细胞肥大及细胞外基质合成的影响.实用儿科临床杂志, 2006, 21(5):268-270
[12]肖朝华,周建华,吴衡生.多球壳菌素对肾小球系膜细胞肥大及周期蛋白的影响.中华肾脏病杂志,2006, 22(8):507-509
[13] Yang N, Isbel NM, Nikolic-Paterson DJ, et al. Local macrophage proliferation in human glomerulonephritis.Kidney Int, 1998, 54:143 ~ 154.
[14] Kang DH , Joly AH , Oh SW ,et al . Impaired angiogenesis in the remnant kidney model (1):Potential role of vascular endothelial growth factor and thrombosponcin-1 . J J Am Soc Nephrol. 2001, 12: 1434 ~1447.
[15] Ohashi R , Kitamura H , Yamanaka N . Peritubular capillary injury during the progression of experimental glomerulonephritis in rats [J] . J Am Soc Neprol , 2000 , 11 (2) : 47~49
[16] Yang N, Isbel NM, Nikolic-Paterson DJ, et al. Local macrophage proliferation in human glomerulonephritis.Kidney Int, 1998, 54:143 ~ 154.
[17]章斌,王伟铭,周同等,肾小管周围毛细血管减少与肾小管间质损伤的关系.中国微循环. 2004 , 8 (5) 279~282.
[18] EI Nahas AM , Muchaneta-Kubara EC , Essawy M , et al . Renal fibrosis : insights into pathogenesis and treatment . Int J Biochem Cell Biol , 1997 , 29:55-62
[19] Eddy AA . Molecular basis of renal fibrosis . Pediatr Nephrol , 2000,15:290-301
[20] Eddy AA. Expression of genes that promote renal interstitial fibrosis in rats with proteinuria . Kidney Int Suppl . 1996,54:S49-54
[21] Fine LG , Orphanides C , Normal JT . Progressive renal disease . The chronic hypoxia hypothesis . Kidney Int , 1998 , 65:S74
[22] Kang DH , Anderson S , Kim YG , et al . Impaired angiogenesis thrombospondinl in renal disease . Am J Kidney Dis , 2001,37(3) : 600-11
[23]邓跃毅,陈以平,贺学林,等.冬虫夏草制剂延缓慢性肾衰竭的机理研究.中国中西医结合肾病杂志.2001,2(7):381-383
[24]孙尔维,张雷.一种新型免疫抑制剂-FTY720.国外医学免疫学分册. 2000,23(1):22-25
[25] Klahr S, Morrissey J, et al Obstructive nephropathy and renal fibrosis .Am J Physiol Renal Physiol , 2002, 283(5) : 861-875
[26] Yang , Jun Wei , Liu You Hua . Blockage of tubular epithelial to myofibroblast transition by hepatocyte growth factor prevents renal interstitial fibrosis [J] . J Am Soc Nephrol , 2002 , 13(1): 96-107
[27] Truong . Cell apoptosis and proferation in experimental chronic obstructive uropathy [J]. Kidney Int , 1996 , 50:200-207
[28] Strutz F , Neilson EG . The role of lymphocytes in the progression of interstitial disease . Kidney Int , 1994 , 45(Suppl 45):S106-S110
[29] Postlethwaite AE , Holness MA , Katai H , et al . Human fibroblasts synethesize elevated levels of extracellular matrix proteins in response to interleukin 4. J Clin Invest , 1992,90:1479-1485
[30] Ricardo SD , Diamond JR . The role of macrophages and reactive oxygen species in experimental hydronephrosis . Semin Nephrol , 1998,18:612-621
[31] Suleiman M, Cury PM , Pestana JO ,et al. FTY720 prevents renal T-cell infiltration after ischemia/reperfusion injury . 2005 , 37(1):373-4
[32] Okada H , Strutz F , Danoff TM , et al . Possible pathogenesis of renal fibrosis . Kidney Int , 1996 , 49(Suppl 54):S37-38
[33] Kalluri R , Neilson EG . Epithelial– mesenchymal transition and its implications for fibrosis . J Clin Invest , 2003 , 112:1773-1784
[34] Klahr S , Morrissey J . Obstructive nephropathy and renal fibrosis [J] . Am J Physiol Renal Physiol , 2002, 283(5):861-875
[35] Fine LG , Orphanides C , Normal JT . Progressive renal disease . The chronic hypoxia hypothesis . Kidney Int , 1998 , 65:S74
[36] Fine LG , Orphanides C , Norman JT , et al. Progressive renal disease: the chronichypoxia hypothesis [J] . Kidney Int . 1998 , 65: 74-78
[37] Zhang B , Wang WM , Shi H . Effects of valsartan and ramiprilon on the peritubular microangium in rats with 5/6 subtotal-nephrectomy [J]. J Nephrol Dialy Transplant , 2003 , 12 (4) : 28-31.
[38] Brinkmann V , Pinschewer DD , Feng L , et al . Fty720: altered lymphocyte traffic results in allograft protection . Transplantation , 2001, 72(5) : 764-769
[39] Im DS . Linking Chinese medicine and G-protein-coupled receptors. Trends in Pharamacological Science , 2003, 24(1):2-4
[40] Nagahara Y , Ikekita M , Shinomiya T . Immunosuppressant FTY720 induces apoptosis by direct induction of permeability transition and release of cytochrome from mitochondria [J] . Immunol , 2000 , 165 : 3250-60
[1] Marcussen N , Nyengaard J , Christensen S. Compensatory growth of glomeruli is accomplished by an increased number of glomerular capillaries. Lab Invest , 1994 ,70 :868
[2] Kang DH , Hughes J , Mazzali M, et al.Impaired angiogenesis in the remnant kidney model ( II) : VEGF administration reduces renal fibrosis and stabilizes renal function1 J Am Soc Nephrol ,2001 , 12 :1448
[3] Murohara Y, Horowitz JR , SliverDR , et al.Vascular endothelial growth factor/ vascular permeability factor enhances vascular permeability via nitric oxide and prostacylin1Circulation ,1998 ;97 :99
[4] Thomas S , Vanuystel J , Gruden G, et al. Vascular endothelial growth factor receptors in human mesangium in vitro and in glomerular disease. J Am Soc Nephrol ,2000 , 11 :1236
[5] Yamagishi S , Inagaki Y, Okamoto T , et al. Advanced glycation end product induced apoptosis and overexpression of vascular endothelial growth factor and monocyte chemoattractant protein-1 in human-cultured mesangial cells. J Biol Chem,2002 , 277 :203091
[6] Schweda F , Blumberg FC , Schweda A , et al. Effects of chronic hypoxia on renal PDGF-A , PDGF-B , and VEGF gene expression in rats. Nephron ,2000 ,86 :161
[7] Levy AP. A cellular paradigmfor the failure to increase vascular endothelial growth factor in chronically hypoxic states.Coron Artery Dis , 1999 , 10 :427
[8] Ferrara N . Role of vascular endothelial growth factor in the regulation of angiogenesis. Kidney Int ,1999 ,56 :794
[9] Williams B , Baker A , Gallacher B , et al. Angiotensin II increases vascular permeability factor gene expression by human vascular smooth muscle cells. Hypertension ,1995 ,25 :913
[10] Cha DR , Kim NH , Yoon JW, et al.Role of vascular endothelial growth factor in diabetic nephropathy.Kidney Int ,2000 (Suppl) ,77 :S104
[11] Kang DH , Nakagawa T , Feng L , et al . Nitric oxide modulates vascular disease in the remnant kidney model.AmJ Pathol ,2002 ,161(1) :239
[12] Smith LE , Shen W, Perruzzi C , et al.Regulation of vascular endothelial growth factor-dependent retinal neovascularization by insulin-like growth factor-1 receptor. Nat Med ,1999 ,5 :1390
[13] ZeisbergM, ,Strutz F , Muler GA1 Renal fibrosis : an update .Curr Opin in Nephrol and Hypertens ,2001 ,10 :315
[14] Kang DH , Joly AH , Oh SW, et al.Impaired angiogenesis in the remnant kidney model (Ⅰ) : Potential role of vascular endothelial growth factor and thrombospondin-1. J Am Soc Nephrol ,2001 ,12 :1434
[15] Eremina V , Haigh J , Nagy A , et al. The role of VEGF-A in glomerular angiogenesis [ abstract ].J Am Soc Nephrol ,2002 ,13 :100A
[16] Horita Y, Miyazaki M, Koji T , et al. Expression of vascular endothelial growth factor and its receptors in rats with protein overload nephrosis. Nephrol Dial Transpl ,1998 , 13 :2519
[17] Grone J-J , Simon M, Grone EF. Expression of vascular endothelial growth factor in renal vascular disease and renal allografts. J Pathol , 1995 ,177 :259
[18] Klanke B , Simon M, Rockl W, et al. Effects of vascular endothelial growth factor (VEGF) / vascular permeability factor (VPF) on haemody namics and permselectivity of the isolated perfused rat kidney. Nephrol Dial Transplant ,1998 ,13 :875
[19] Celletti FL , Waugh JM, Amabile PG, et al. Vascular endothelial growth factor enhances atherosclerotic plaque progression. Nat Med , 2001 , 7 :425
[20] Flyvbjerg A , Dagnaes-Hansen F , De Vriese AS , et al . Amelioration of long-term renal changes in obese type 2 diabetic mice by a neutralizing vascular endothelialgrowth factor antibody. Diabetes ,2002 ,51 (10) : 3090-4
[21] El Awad B , Kreft B , Wolber EM , et al . Hypoxia and interleukin-1beta stimulate vascular endothelial growth factor production in human proximal tubular cells . Kidney Int , 2000 Jul ; 58(1) : 43-50
[22] Kanellis J , Frasre S , Katerelos M , et al . Vascular endothelial growth factor is a survival factor for renal tubular epithelial cells .Am J Physiol Renal Physiol , 2000; 278(6): F905-15
[23] Choi YJ , Chakraborty S , Nguyen V , et al . Peritubular capillary loss is associated with chronic tubulointerstitial injury in human kidney : altered expression of vascular endothelial growth factor .Hum Pathol , 2000; 31(12): 1491-7
[24] Cosio F, Alamir A , Yim S , et al . Patient survival after renal transplantation:Ⅰ.The impact of dialysis pre-transplant . Kidney Int , 1998;53(3): 767-72
[25] Fontan M , Rodriguez Carmona A , Garcia Falcon T , et al . Perit Dial Int , 1996; 16:48-51
[26] Ito F , Nakazawa H , Rvoji O , er al . Cytokines accumulated in acquired renal cysts in long term hemodialysis patients. Urol Int . 2000;65(1): 21-7
[27] Ojo AO, Wolfe RA , Held PJ, et al. Delayed graft function : risk factors and implications for renal allograft survival .Transplantation , 1997;63(7):968-74
[28] Kang DH , Kim YG , Andoh TF ,et al . Post-cyclosporine-mediated hypertension and nephropathy: amelioration by vascular endothelial growth factor . Am J Physiol Renal Physiol , 2001 ; 280(4) : F727-36
[29] Pilmore HL , Eris JM , Painter DM , et al . Vascular endothelial growth factor expression in human chronic renal allograft rejection . Transplantation . 1999 ; 67(6) 929-33
[30] Nakamoto Y , Imai H , Yasuda T , et al . A spectrum of clinicpathological features of nephropathy associated with POEMS syndrome . Nephrol Dial Transplant . 1999;14(10):2370-8
[31] Lee JS , Kim HS , Jung JJ , et al . Expression of vascular endothelial growth factor in renal cell carcinoma and the relation to angiogenesis and P53 protein expression . J Surg Oncol .2001; 77(1):55-60
[32] Drevs J , Hofmann I , Hugenschmidt H , et al . Effects of PTK787/ZK 222584 , a specific inhibitor of vascular endothelial growth factor receptor tyrosine kinases , on primary tumor , metastasis , vessel density , and blood flow in a murine renal cell carcinoma model . Cancer Res . 2000;60(17):4819-24
[33] Kuroda N ,Moriki T ,Komatsu F ,et al .Adult-onset giant juxtaglomerular cell tumor of the kidney. Pathol Int ,2000 ;50 (3) : 249～54
[33] Masuda Y, Shimizu A , Mori T , et al. Vascular endothelial growth factor enhances glomerular capillary repair and accelerates resolution of experi2 mentally induced glomerulonephritis1 AmJ Pathol ,2001 ,159 :599
[34]. Kim NH , Oh JH , Seo JA , et al . Vascular endothelial growth factor (VEGF) and soluble VEGF receptor FLT-1 in diabetic nephropathy . Kidney Int . 2005;67(1):166-67
[35] Harada M , Mitsuyama K , Yoshida H , et al . Vascular endothelial growth factor in patients with rheumatoid arthritis.Scand J Rheamatol .1998;27(5):377-80