共刺激信号阻断剂基因局部转染对大鼠移植肾存活的影响

目的　观察CTLA 4Ig基因局部转染延长移植肾存活的效能。 方法　以CTLA 4Ig基因重组腺病毒为载体 ,将CTLA 4Ig基因转入BN大鼠肾脏。以BN大鼠为供者 ,Lewis大鼠为受者 ,行同种肾移植术。用经CTLA 4Ig基因转染的供肾移植给受者为转染组 ;用未转染CTLA 4Ig基因的供肾移植给受者为对照组。观察移植肾存活时间和术后肾功能变化。结果　转染组移植肾存活 (32± 8.0 )d ,对照组移植肾存活 (8.5± 1.4 )d ,转染组存活时间明显延长 ;转染组术后血清肌酐较同期对照组明显为低。结论　CTLA 4Ig基因局部转染供肾可明显延长移植肾的存活时间。     

小鼠同种异体胰岛移植排斥反应中可诱导共刺激分子的表达及阻断其对排斥反应的影响

移植胰岛长期存活率不高，主要原因是移植术后排斥反应，共刺激信号在排斥反应的发生中起重要作用。本实验旨在观察共同刺激分子-可诱导其刺激分子（ICOS）是否参与了小鼠同种异体胰岛移植排斥反应的发生，并用ICOS单克隆抗体（ICOSmAb）阻断ICOS，研究其对排斥反应的抑制作用。[第一段]     

可诱导共刺激分子基因表达抑制降低大鼠T淋巴细胞活化增殖能力的研究

目的 RNAi抑制大鼠T淋巴细胞可诱导共刺激分子(ICOS)基因表达,观察其对T淋巴细胞功能的影响.方法 选择4个干扰位点,插入pSilencer 4.1-CMV neo载体,转染大鼠淋巴细胞.RT-PCR检测ICOS基因的抑制程度,流式检测ICOS表达变化.MLR检测细胞的增殖能力,ELISA法检测细胞因子IFN-γ和IL-4分泌水平.结果 转染后,T淋巴细胞ICOS的mRNA和ICOS分子表达水平下降(P＜0.05),淋巴细胞增殖能力明显降低,同时IFN-γ和IL-4的水平也减低.结论 大鼠淋巴细胞ICOS基因的表达抑制,降低了T淋巴细胞活化增殖能力.     

大鼠慢性移植肾肾病动物模型的制作经验总结

目的 总结制作稳定的大鼠慢性移植肾肾病(CAN)动物模型的经验.方法 以F344近交系大鼠为供者,取供者左肾作为供肾,原位低温灌注;以Lewis近交系大鼠为受者,行左侧原位肾移植,供肾动脉与受者腹主动脉端侧吻合,供肾静脉与受者肾静脉端端吻合,输尿管带膀胱瓣与受者膀胱吻合.术后用环孢素A灌胃10 d,剂量为10 mg·kg-1·d-1.每月采集受者血液和尿液,测定血肌酐及24 h尿蛋白,分别于术后2、4个月获取移植肾进行病理检查.结果 45只进行移植,手术成功率为85%,单次手术时间为(120±20)min.移植后1个月,大鼠即出现血肌酐、尿素氮及血胱抑素升高,24 h尿蛋白增加,与术前相比,各项指标均升高(P<0.05);术后2个月及4个月,除尿蛋白继续增加外,其余观察指标上升不明显.移植术后2个月,移植肾有轻度至中度的间质纤维化,淋巴细胞和浆细胞的浸润;4个月时,移植肾可见广泛的间质纤维增生,间质细胞大量浸润,肾小球基底膜增厚、硬化、闭塞,肾小管萎缩退化,符合CAN的病理改变.结论 通过充分的手术强化训练及改进,规范大鼠取、肾、移植术中、术后管理的每个细节,大鼠CAN模型的成功率及稳定性高.

Abstract：

Objective To summarize the experience of establishing the stable rat model of chronic allograft nephropathy. Methods We used Fisher rats as donors and Lewis rats as recipients.After the left kidney of the donor perfused in situ under hypothermic condition, the left renal vein,abdominal aorta and bladder flap of the donor was anastomosed with the left renal vein, renal artery and bladder of the recipient, respectively. The recipients were given cyclosporin oral solution 10 mg/kg every day by gavage for 10 days after transplantation. The blood and urine samples were collected 1 month, 2 months and 4 months after transplantation and renal function and total urine protein were examined. The pathological changes of the renal allograft were observed 2 and 4 months after transplantation. Results Forty-five rats received operation and achievement ratio was 85%. The renal transplantations were finished in 120 ± 20 min. The Scr, BUN, Cycs and total urine protein demonstrated a significant increase one month after transplantation. On the second and fourth month,with the exception of urine protein continued to increase, the other indicators did not change significantly. Two months after transplantation renal pathology demonstrated light to moderate interstitial fibrosis, infiltration of lymphocytes and plasma cells. At 4th month the renal allografts showed extensive interstitial fibrosis, a large number of infiltrating interstitial cells, thickening,hardening, occlusion of glomerular basement membrane, and renal tubular atrophy that were consistent with pathological changes of chronic allograft nephropathy. Conclusion Through adequate surgical training and improvement, and specification for rat nephrectomy, transplantation surgery,and postoperative management in every detail, the model with high success rate and stability can be achieved.     

慢性移植肾肾病研究进展

本文主要阐述了慢性移植肾肾病的发病机制、诊断和治疗方面研究现状及进展。     

大豆异黄酮对大鼠慢性移植肾肾病的防治作用

目的通过对肾移植大鼠的饮食营养干预，观察大豆异黄酮对慢性移植肾肾病的防治作用。方法选择近交系雄性Fisher（F344）大鼠作为供者，雄性Lewis（Lew）大鼠作为受者，采用显微外科技术制作肾移植模型。将受者随机分为三组，分别给予高异黄酮大豆蛋白饲料（HIS组）、低异黄酮大豆蛋白饲料（LIS组）或酪蛋白饲料（CAS组）。移植前和移植后第4、12和24周时检测血压，并收集受者的血和尿样，检测尿蛋白和血肌酐含量。24周时处死大鼠获取移植肾，行组织学和免疫组织化学检查。结果在移植后4周时，HIS组受者的尾动脉收缩压、24h尿蛋白含量和血肌酐浓度即低于LIS组和CAS组，但差异无统计学意义（P〉0．05）；移植后12周和24周时，HIS组的受者尾动脉收缩压、24h尿蛋白含量和血肌酐浓度均较LIS组和CAS组显著降低（P〈0．05）；移植后24周时，HIS组移植肾组织的间质纤维化和炎症、血管硬化、肾小球硬化和肾小管萎缩等慢性病变均较LIS组和CAS组为轻（P〈0．05）；HIS组移植肾组织中转化生长因子-β1（TGF-β1）的表达和分泌均较LIS组和CAS组为少（P〈0．05）。结论大豆异黄酮对移植肾功能和结构有保护作用，可作为一种防治慢性移植肾肾病的新方法。     

慢性移植肾肾病的治疗方法探讨

目的：探讨治疗慢性移植肾肾病(CAN)的有效途径。方法：按照不同治疗方案将CAN234例患者分为4组，比较各组的治疗效果。结果：234例的总有效率为34．2％，4组的有效率分别为19．3％、32．6％、46．9％、42．9％结论：随着免疫抑制方案的不同，临床上所见慢性排斥发生机制也有差别，治疗上宜根据患者具体情况，采取相应的综合措施，抗病毒、降血脂、降血压、治疗糖尿病等，可取得一定效果；结合传统中药治疗，可明显提高有效率。     

Smad7及Foxp3分子在慢性移植肾肾病检测中的临床应用

目的：研究Smad7和Foxp3在慢性移植肾肾病（CAN）中的作用以及两指标在该疾病中的潜在联系.方法：应用ELISA法和原位杂交技术分别检测32例不同病理分级CAN（其中CAN Ⅰ级10例,Ⅱ级9例,Ⅲ级13例）患者外周血中及活检标本当中的Smad7和Foxp3水平,另取9例移植后肾功能正常志愿患者的活检标本和外周血标本做为对照.结果：CAN各级的Smad7和Foxp3表达水平无论是外周还是活检标本当中皆明显低于对照组的表达水平（P＜0.05）,而CANⅠ级与CANⅡ级差异无统计学意义（P＞0.05）,CAN Ⅰ级和CANⅡ级对比CANⅢ级的表达水平差异有统计学意义S（P＜0.05）,Smad7和Foxp3的表达有明显联系（P＜0.05）,而有3例标本两者的表达差异较大.结论：移植肾活检穿刺能客观的反映CAN的病理变化,而外周血当中的Smad7和Foxp3水平基本能够反映CAN的发展程度.是检测CAN的一个不错的无创指标,Smad7和Foxp3对慢性移植肾肾病的发生发展起重要作用,而Smad7可能通过某种机制调控Foxp3的表达.     

川芎嗪联合丹参延缓肾移植大鼠慢性移植肾肾病进程的作用

目的 研究川芎嗪(TMP)联合丹参延缓肾移植大鼠慢性移植肾肾病进程的作用及机制.方法 以Fisher 344大鼠和Lewis大鼠分别作为肾移植的供、受者进行原位肾移植,并建立CAN模型.按随机数字表法将受鼠分为5组:环孢素A(CsA)组(A组),TMP+ CsA组(B组)、丹参+ CsA组(C组)、TMP+丹参+CsA组(D组)及空白对照组(E组,术使用任何药物).分别于术后2、4、6、8和12周,处死每组5只受鼠,取移植肾进行移植肾组织病理学变化,采用免疫组织化学法检测移植肾组织转化生长因子β1(TGF-β1)的表达,采用荧光定量聚合酶链反应法检测移植肾组织TGF-β1 mRNA的表达.结果　术后空白对照组受鼠的存活时间均未超过2周.A组于术后4周时最早出现CAN病理改变,B组和C组出现CAN病理改变的时间较A组晚,D组出现CAN病理改变的时间最晚,且病理改变程度最轻.术后各组Banff评分均呈现明显的上升趋势,相同时间点,A组明显高于其他3组(P＜0.05和P＜0.0 1),D组明显低于与B组和C组(P＜0.05),而B组与C组间无显著差异(P＞0.05).随着术后时间的推移,各组TGFβ1表达强度呈逐渐增加的趋势,相同时间点,A组TGF-β1表达强度最高,与其他3组比较,差异均有统计学意义(P＜0.05和P＜0.01),D组明显低于B组和C组(P＜0.05),B组与C组间无明显差异(P＞0.05).各组TGF-β1 mRNA表达的变化趋势及组间差异与TGF-β表达强度的情况相一致.结论 TMP或丹参均具有通过下调TGF-β1 的表达延缓肾移植大鼠CAN进程的作用,当两者联合应用时作用更为明显.     

慢性移植肾肾病

目前，肾移植的近期(术后1年内)效果虽已显著改善，但远期效果却未能得到相应的提高，移植肾的平均半寿期仍徘徊在7～10年，这主要是因为不少患者在肾移植后数月、特别是1年或数年后，移植肾功能进行性减退(常伴有高血压、蛋白尿等)，最终导致移植肾功能衰竭。是什么原因导致了这一现象，至今尚不清楚。病理研究发现，这类移植肾表现为明显的瘢痕化趋势，在组织学上移植物呈现出以细胞外基质沉积、间质纤维化、肾小管萎缩为特点的各种病理改变，然而这些改变都是非特异性的，许多免疫和非免疫因     

慢性移植肾肾病模型中骨桥蛋白、α平滑肌肌动蛋白和E钙黏蛋白的表达

目的 观察骨桥蛋白(OPN)、α平滑肌肌动蛋白(α-SMA)、E钙黏蛋白在大鼠慢性移植肾肾病(CAN)模型移植肾中的表达及其表达的相关性.方法 以Fisher大鼠为供者,Lewis大鼠为受者,进行原位异体肾移植(CAN组);以供、受者均为Lewis大鼠的肾移植模型作为对照.术后12周处死受者,取血、尿样本检测移植肾功能,取移植肾进行HE染色和天狼星红染色,观察其组织形态学变化.采用免疫组织化学法和免疫印迹法观察移植肾组织OPN、α-SMA和E钙黏蛋白的表达.结果 CAN组大鼠移植肾组织形态学符合CAN的病理改变.免疫组化结果显示,与对照组相比较,CAN组肾小管上皮及肾间质中OPN和α-SMA的表达增强,而CAN组小管上皮E钙黏蛋白的表达明显下降.免疫印迹结果与免疫组化结果一致,CAN组移植肾组织中OPN和α-SMA的表达相对灰度值分别为85.74±2.29和88.79±4.44,对照组分别为14.25±0.71、11.21±0.56,两组间的差异有统计学意义(P＜0.05).CAN组E钙黏蛋白的相对灰度值为24.96±0.02,对照组为75.04±3.21,两组间的差异有统计学意义(P＜0.05).两组大鼠移植肾OPN的表达量与α-SMA的表达量呈正相关(r=0.746,P＜0.05),与E钙黏蛋白的表达量呈负相关(r=-0.526,P＜0.05).结论 CAN大鼠移植肾中OPN的表达增加,OPN可能参与了移植肾肾小管上皮细胞向间充质细胞转化的过程.

Abstract：

Objective To investigate the expression of OPN, α-SMA, E-cadherin and their correlation in the chronic allograft nephropathy (CAN) rat model, and to explore the possible role of OPN in CAN.Methods Orthotopic renal-transplantation using Fisher rats as donors and Lewis rats as recipients was done to establish CAN group, and Lewis to Lewis rats as control group. Rats in each group were sacrificed 12 weeks after the surgery. Blood and urine were collected for further test. Allograft samples were collected and sectioned for HE, Sirus-red staining, immunohistochemistry and Western blot.Results There were CAN morphological changes of the allograft in CAN group. As compared with control group, immunohistochemistry and Western blot revealed that the expression of OPN and α-SMA in CAN group was significantly increased, and that of E-Cadherin reduced. Its trend was correlated with the inflammatory response and the EMT of tubule epithelial cells.Conclusions OPN expression in rat CAN model is significantly up-regulated. OPN may play a role in CAN. OPN might affect the CAN by promoting EMT of tubule epithelial cells.     

磷酸化肌球蛋白轻链在大鼠慢性移植肾肾病早期病变中的作用及其机制

目的 探讨磷酸化肌球蛋白轻链(pMLC)在慢性移植肾肾病(CAN)中的作用与机制.方法 依照标准的CAN大鼠模型进行左肾原位移植,受者为LEW大鼠,供者为F344大鼠,另取雄性F344大鼠和LEW大鼠仅行单肾切除术,分别作为对照.分别于术后4、8及12周时,收集各组大鼠的24 h尿量,并检测其尿肌酐水平,计算肌酐清除率.然后取各组大鼠血液标本,测定血清肌酐水平.采用Banff分级标准评定各组肾小球肾病、肾小管萎缩、间质纤维化及血管内膜增厚程度.采用免疫组织化学法和采用蛋白质印迹法检测肾组织中磷酸化MLC(pMLC)和整合素连接激酶(ILK)的表达部位及表达水平.结果 移植组大鼠术后4周时肾间质可见单个核细胞浸润,12周时可见血管平滑肌细胞的移行与增殖.移植组大鼠各时相肾组织pMLC和ILK表达水平显著高于Lewis对照组及F344对照组,且随着移植时间的延长有逐渐增高趋势.大鼠移植肾组织中pMLC表达水平与24 h尿蛋白定量、血清肌酐水平、肾间质单个核细胞浸润、肾小动脉血管平滑肌细胞数量、Banff评分等呈显著正相关,相关系数(r)分别为0.273(P＜0.05)、0.434(P＜0.01)、0.525(P＜0.01)、0.676(P＜0.01)、0.570(P＜0.01),在移植后4周时,pMLC表达水平与肾小管间质ILK表达水平呈显著正相关,r=0.778(P＜0.01).结论 pMLC在慢性移植肾病早期病理变化中发挥重要作用,移植肾肾小管间质及肾小动脉中pMLC表达上调与ILK的作用机制相关.

Abstract：

Objective To investigate the role and mechanism of phosphate myosin light chain (pMLC) in the rat kidney of chronic allograft nephropathy (CAN) model. Methods The left donor kidneys from Fisher (F344) rats were orthotopically transplanted into Lewis recipients. Meanwhile, the F344 rats and LEW rats with resection of the right kidney served as control groups. Animals were harvested respectively at the 4th, 8th and 12th week after transplantation. The creatinine clearance rate (CCr) was calculated by urine creatinine of 24-h urine. Blood samples were collected from rats for determination of serum creatinine. The expression of pMLC was detected by using Western blotting and immunohistochernistry, and that of integrin-linked kinase (ILK) by using immunohistochemistry. Results Mononuclear cells infiltration of allografts was markedly aggravated as compared to the controls. Allografts got severe interstitial fibrosis and tubular atrophy at 12th week after transplantation. The expression of pMILC and ILK was up-regulated in the kidney of CAN rats after transplantation, and increased more significantly as the time went on. The expression of pMILC was significantly correlated with 24-h urine protein excretion (r= 0. 273, P＜0. 05), serum creatinine levels (r = 0. 434, P＜0. 01 ), the number of tubulointerstitial infiltrated mononuclear cells (r = 0. 525, P＜0. 01 ), the number of smooth muscle cells (SMC) in vascular wall (r= 0. 676, P＜0. 01 ) and the extent of interstitial fibrosis (r= 0. 570, P＜0. 01 ).There was a significantly positive correlation between ILK and pMLC in CAN rats at the 4th week after transplantation (r= 0. 778, P＜0. 01 ). Conclusion pMLC might play an key role in CAN, and the over-expression of ILK might be involve in the pathogenesis of CAN.     

The aetiology and pathogenesis of chronic allograft nephropathy

Renal transplantation is the ultimate form of renal replacement therapy, and is the treatment of choice for many patients with end-stage renal failure. The advent of calcineurin inhibitor based immunosuppression resulted in the 1-year renal allograft failure rate dropping from around 50% twenty years ago to less than 10% in more recent times. Despite a massive improvement in renal allograft survival in the first year following transplantation 10-year graft survival can be as low as 50%. Chronic allograft nephropathy (CAN) is recognised as the main cause of renal allograft failure following the first year after transplantation. The diagnosis of CAN can only be made histologically. Typically biopsy specimens in grafts with CAN demonstrate an overall fibrotic appearance effecting the vascular endothelium, renal tubules, interstitium, and glomerulus. The risk factors for CAN are divided into alloimmune and alloimmune independent. Alloimmune dependent factors include acute cellular rejection, severity of rejection, subclinical rejection and HLA mismatch. Alloimmune independent factors such as delayed graft function, donor age, Cytomegalovirus infection, donor/recipient co-morbidity and of course calcineurin inhibitor toxicity are important in the development of CAN. The pathogenesis of CAN is complex, multifactorial, and unfortunately incompletely understood. There are a number of pivotal steps in the initiation and propagation of the fibrosis seen in biopsy specimens from kidneys with CAN. Endothelial activation in response to one or more of the aforementioned risk factors stimulates leukocyte activation and recruitment. Recruited leukocytes subsequently infiltrate through the endothelium and induce key effector cells to secrete excessive and abnormal extracellular matrix (ECM). Enhanced deposition of ECM is a histological hallmark of CAN. This paper aims to present a concise yet accurate and up-to-date review of the literature concerning the aetiological factors and pathological processes which are present in the generation of CAN.     

西罗莫司与钙调磷酸酶抑制剂联合方案在慢性移植肾肾病中的应用

目的 探讨西罗莫司(SRL)与减低剂量的环孢素A(CsA)或他克莫司(Tac)联合方案在慢性移植肾肾病(CAN)中的应用.方法 53例无特定病因所致的CAN患者,在原CsA(或Tac)+吗替麦考酚酯(MMF)+泼尼松(Pred)的免疫抑制方案上加用SRL(四联方案),其中CsA(或Tac)和MMF的用量减少25％～50％.治疗12个月,观察受者血肌酐、肾小球滤过率(GFR)、血胆固醇、血甘油三酯、尿蛋白等的改变.结果 四联方案治疗前患者血肌酐为(161.51±106.48)μmol/L,治疗后1个月为(138.47±67.74) μmol/L,治疗后6个月为(126.51±56.21)μmol/L,治疗后12个月为(123.43±54.18)μmol/L.与治疗前相比较,治疗6个月和12个月后,差异有统计学意义(P＜0.05,P＜0.01).四联方案治疗前患者GFR为(0.754±0.302) ml/s,治疗后1个月为(0.868±0.358)ml/s,治疗后6个月为(0.952±0.347) ml/s,治疗后12个月为(1.007±0.394) ml/s.治疗6个月和12个月后,患者GFR与治疗前相比较,差异有统计学意义(P＜0.05,P＜0.01).治疗1、6和12个月后,患者血胆固醇和甘油三酯与治疗前相比较,差异均无统计学意义(P＞0.05,P＞0.05).四联方案治疗前患者尿蛋白阳性率为9.4％,治疗后1个月为13.2％,治疗后6个月为22.6％,治疗后12个月为26.4％.治疗12个月后蛋白尿阳性率与治疗前相比较,差异有统计学意义(P＜0.05).结论 应用SRL+ CsA(或Tac)+ MMF+ Pred四联方案改善了CAN患者的血肌酐和GFR,但增加了患者蛋白尿的发生率.     

将钙调磷酸酶抑制剂转换为西罗莫司在慢性移植肾肾病中的临床治疗效果

目的 探讨慢性移植肾肾病(CAN)患者将免疫抑制方案中钙调磷酸酶抑制剂(CNI)转换为西罗莫司(SRL)的有效性及安全性.方法 选取72例经移植肾活检证实发生CAN的受者,其中35例将免疫抑制方案中CNI转换为SRL(SRL组),其余37例继续原CNI方案(CNI组).另取10例因其他原因将CNI转换为SRL治疗的受者,将45例转换为SRL的患者分为A组[血肌酐(SCr)＜120 μmol/L),B组(SCr为120～200 μol/L,且Banff分级为Ⅰ～Ⅱ级),C组(SCr为120～200 μmol/L,且Banff分级在Ⅱ级以上),D组(SCr＞200 μmol/L).随访期为24个月,检测各组随访期内的各临床指标.结果 转换治疗前,两组间SCr和肾小球滤过率(eGFR)的差异无统计学意义(P＞0.05);转换治疗后24个月内,SRL组SCr水平和eGFR较CNI组明显改善(P＜0.05),而CNI组的移植肾功能有逐渐衰退的趋势.SRL组尿蛋白及血脂明显上升(P＜0.05),而CNI组变化不大;SRL组血小板计数较CNI组明显下降(P＜0.05),两组间其他指标的差异无统计学意义(P＞0.05).A组患者各指标在转换治疗前后的变化并不大,B组患者的肾功能及蛋白尿有改善明显,C组和D组患者肾功能有不同程度衰退情况,且蛋白尿加重.结论 SRL转换治疗对于稳定及改善CAN患者的移植肾功能是有效、安全的,CAN早期进行转换(SCr＜200 μmol/L)效果明显.     

沙利度胺对大鼠慢性移植物血管病的保护作用

目的探讨沙利度胺对大鼠慢性移植物血管病的保护作用。方法实验分为4组：同种移植对照组,供、受者均为Lewis大鼠,无特殊处理;异种移植对照组,Lewis大鼠接受Brown-Norway（BN）大鼠腹主动脉移植（BN-Lewis）;沙利度胺小剂量组（50mg·kg-1·d-1,BN-Lewis）;沙利度胺大剂量组（100mg·kg-1·d-1,BN-Lewis）。于移植后60d取移植动脉,分别进行组织病理学观察、测量内膜厚度,免疫组织化学法和蛋白质印迹法检测TNF-α及上皮细胞增殖细胞核抗原（PCNA）在移植动脉中的表达。结果同种移植对照组移植动脉形态正常;异种移植对照组移植动脉呈移植物血管病表现,血管内膜增厚;沙利度胺小剂量及大剂量组移植动脉呈内膜炎改变,内膜厚度比异种移植对照组减小（P〈0．05）。与异种移植对照组相比,沙利度胺小剂量组和大剂量组移植动脉TNF-α及PCNA蛋白表达量降低（均P〈0．05）。结论沙利度胺能降低移植动脉TNF-α及PCNA蛋白的表达,缓解移植动脉的纤维化进程以及内膜增生,对慢性排斥反应所致的移植物血管病具有抑制作用。     

肾小管上皮细胞转分化在慢性移植肾肾病中的作用

目的探讨肾小管上皮细胞-肌成纤维细胞转分化在慢性移植肾肾病(CAN)的发生、发展中的作用。方法36例移植肾穿刺标本分为正常组(N组),CAN组(C组),依Banff 97标准将C组按CANⅠ、Ⅱ、Ⅲ级分为C_1、C_2、C_3组,每组9例。免疫组化染色半定量分析比较α-平滑肌肌动蛋白(α-SMA)、波形蛋白(VIM)、角质蛋白(CK_(AEI/AE3))和转化生长因子β1(TGF-β1)在各组中的表达,线性相关分析TGF—β1和α-SMA、VIM、CK_(AE1/AE3)表达的相关性。结果C_1、C_2、C_3组α-SMA表达积分分别为0．95±0．07、1．78±0．12、2．42±0．31,VIM表达积分分别为0．74±0．05、1．31±0．18、2．34±0．25,组间呈递增趋势,均明显高于N组α-SMA表达积分0．07±0．02、VIM表达积分0．09±0．02(P＜0．05)；移植肾组织中TGF-β1表达与α-SMA、VIM呈正相关(r分别为0．73、0．68,P＜0．05),而与CKAE1/AE3表达呈负相关(r=-0．71,P＜0．05)。结论肾小管上皮细胞-肌成纤维细胞转分化参与了CAN的发生、发展,而局部肾组织中TGF-β1的表达上调可能是介导肾小管上皮细胞转分化的重要原因。     

Vascular endothelial growth factor in chronic rat allograft nephropathy

BACKGROUND: Chronic allograft nephropathy (CAN) is a complex process of alloimmune responses and chronic inflammation leading to fibrosis and vasculopathy. We examined the biological role of proinflammatory vascular endothelial growth factor (VEGF) in a rat renal transplantation model of CAN. METHODS: Syngraft and allograft recipients were treated with a suboptimal dose of cyclosporine A which allows acute rejection and CAN to develop. Intragraft VEGF, VEGFR-1 and VEGFR-2 expressions were determined at 5, 14, 30 and 60 days. Protein tyrosine kinase inhibitor PTK787 was used to inhibit VEGFR activity. RESULTS: In nontransplanted kidneys and syngrafts, mild VEGF expression was observed in the glomeruli and tubuli. VEGFR-1 was detected in vascular structures and VEGFR-2 in glomeruli as well. In allografts, total intragraft VEGF expression and interstitial inflammatory cell VEGF expression were induced and correlated with the chronic allograft damage index (CADI) score. Total intragraft and interstitial inflammatory cell VEGFR-1 expression was induced and interstitial cell VEGFR-1 expression correlated with the CADI score. Blocking VEGF receptor signaling with PTK787 significantly reduced fibrosis and the CADI score, but did not affect early inflammation or VEGF, VEGFR-1, VEGFR-2 expressions compared to vehicle treated group. CONCLUSIONS: Interstitial inflammatory cell VEGF and VEGFR-1 expressions are induced during the development of CAN. Increased VEGF activity may enhance the alloimmune induced inflammatory responses leading to fibrosis and CAN.