【摘要】 目的探讨缺血再灌注损伤致慢性移植肾肾病(CAN)的机理及药物干预的防治效果和作用机制。方法利用已建立的大鼠原位异体肾移植硬化加快模型进行原位肾移植。实验分组:A组单用环孢素A(CsA)10mg/(kg·d),B、C、D组分别给予小剂量4mg/(kg·d)、大剂量8mg/(kg·d)益生注射液或吗替麦考酚酯MMF,20mg/(kg·.d),联合应用CsA10mg/(kg·d),E组为空白对照。移植术后4、8及12周处死动物,观察移植肾病理变化并测定肾功能。采用免疫组化和实时荧光定量PCR检测各组TGF-β1、Smad2、Smad7蛋白和mRNA表达及其在移植肾的定位情况。结果大剂量益生组,血浆肌酐水平升高较小剂量组低,CAN病理改变较小剂量组为轻。移植肾标本中,A、B组Smad2、TGF-β1表达上调、Smad7表达下调,尤以A组明显,A组与其它各组相比在各时间点上差异均有统计学意义(P<0.05);移植肾肾小管上皮细胞、间质细胞及肾小球,Smad2、TGF-β1表达均呈强阳性,Smad7表达呈弱阳性;C、D组Smad2、TGF-β1表达下调,而Smad7表达上调,组间差异无统计学意义。结论缺血再灌注损伤致CAN机理可能是通过TGF-β1过表达及上调Smad2表达,下调Smad7表达,直接或间接致移植肾实质和血管的纤维化。中药益生注射液及MMF可能通过下调TGF-β1及Smad2表达、上调Smad7表达而发挥其抗纤维化作用。更多还原

【Abstract】 Objective This study is aiming to investigate the mechanism and drug intervention of chronic allograft nephropathy (CAN) induced by renal ischemia-reperfusion injury. Methods The closed colony strain Sprague-Dawley (SD) and Wistar Rats were used as donor and recipient, respectively. Orthotopic kidney transplantation was performed following the procedure of our previous study. The rats were divided into five groups: Group A only received CsA with 10 mg/(kg·d); except CsA, Group B,C,D received Yi Sheng injection with 4 mg/(kg·d), 8 mg/(kg·d), and MMF 20 mg/(kg·d), respectively. Group E was control group. According to Banff standard, the serum creatinine (SCr) level and pathological change of rat grafted kidney were observed at the 4th, 8th and 12th weeks post-transplantation. The immunohistochemistry and real-time fluorescence quantitative polymerase chain reaction were used to comprehend the localization and expression of TGFβ1 and Smad2, 7 in the transplant kidney. Results Compared with the lower dosage group, the differences of SCr level and pathological changes of CAN at all the time points after 8th week were statistically significant in the high dosage Yi Sheng group. It was showed that the Yi Sheng injection had the protective effect on CAN with a dose-dependent fashion. After transplantation, the rat kidney-sclerosis model showed that the up-regulated expressions of TGF-β1 and Smad2 and the down-regulated expression of Smad7 in Group A and Group B were statistically significant, which meant that the difference was obvious when Group A compared with the other 4 groups. The expressions of TGF-β1 and Smad2 were strongly positive in tubular epithelial cell, interstitial cell and glomerulus, while the expression of Smad7 was weak. Thickening of endovascular could significantly be inhibited in high dosage of Yi Sheng and MMF group. Conclusion The up-regulated expressions of TGF-β1 and Smad2 and the down-regulated expression of Smad7 may accelerate the progression of CAN alone or with immune factors. The traditional Chinese medicine Yi Sheng injection and MMF can down-regulate the expression of TGF-β1 and Smad2 and block the down-regulated expression of Smad7, which may postpone and lessen the progression of CAN.更多还原