转化生长因子-β抗体可改善单侧输尿管梗阻所致的肾小管凋亡。
Antibody to transforming growth factor-beta ameliorates tubular apoptosis in unilateral ureteral obstruction.
作者信息
Miyajima A, Chen J, Lawrence C, Ledbetter S, Soslow R A, Stern J, Jha S, Pigato J, Lemer M L, Poppas D P, Vaughan E D, Felsen D
机构信息
Center for Pediatric Urology and Laboratory for Minimally Invasive Urologic Surgery, New York, New York, USA.
出版信息
Kidney Int. 2000 Dec;58(6):2301-13. doi: 10.1046/j.1523-1755.2000.00414.x.
BACKGROUND
Unilateral ureteral obstruction (UUO) is characterized by progressive renal atrophy, renal interstitial fibrosis, an increase in renal transforming growth factor-beta (TGF-beta), and renal tubular apoptosis. The present study was undertaken to determine the effect of a monoclonal antibody to TGF-beta (1D11) in UUO.
METHODS
Mechanical stretch was applied to tubular epithelial cells (NRK-52E) by a computer-assisted system. Three doses of 1D11 (either 0.5, 2, or 4 mg/rat) were administered to rats one day prior to UUO and every two days thereafter, and kidneys were harvested at day 13. Fibrosis was assessed by measuring tissue hydroxyproline and mRNA for collagen and fibronectin. Apoptosis was assessed with the terminal deoxy transferase uridine triphosphate nick end-labeling assay. TGF-beta levels were determined by bioassay. Western blot and immunostaining were used to identify proliferating cell nuclear antigen (PCNA), p53, bcl-2, and inducible nitric oxide synthase (iNOS).
RESULTS
Stretch significantly induced apoptosis in NRK-52E cells, which was accompanied by an increased release of TGF-beta; 1D11 (10 microg/mL) totally inhibited stretch-induced apoptosis. Control obstructed kidney contained 20-fold higher TGF-beta as compared with its unobstructed kidney; 1D11 neutralized tissue TGF-beta of the obstructed kidney. Control obstructed kidney exhibited significantly more fibrosis and tubular apoptosis than its unobstructed counterpart, which was blunted by 1D11. In contrast, 1D11 significantly increased tubular proliferation. p53 immunostaining was localized to renal tubular nuclei of control obstructed kidney and was diminished by 1D11. In contrast, bcl-2 was up-regulated in the 1D11-treated obstructed kidney. Total NOS activity and iNOS activity of the obstructed kidney were increased by 1D11 treatment.
CONCLUSION
The present study strongly suggests that an antibody to TGF-beta is a promising agent to prevent renal tubular fibrosis and apoptosis in UUO.
背景
单侧输尿管梗阻（UUO）的特征为进行性肾萎缩、肾间质纤维化、肾转化生长因子-β（TGF-β）增加以及肾小管凋亡。本研究旨在确定抗TGF-β单克隆抗体（1D11）在UUO中的作用。
方法
通过计算机辅助系统对肾小管上皮细胞（NRK-52E）施加机械牵张。在UUO前一天给大鼠施用三剂1D11（分别为0.5、2或4mg/大鼠），此后每两天给药一次，并在第13天收获肾脏。通过测量组织羟脯氨酸以及胶原蛋白和纤连蛋白的mRNA来评估纤维化。用末端脱氧转移酶尿苷三磷酸缺口末端标记法评估凋亡。通过生物测定法测定TGF-β水平。使用蛋白质印迹和免疫染色来鉴定增殖细胞核抗原（PCNA）、p53、bcl-2和诱导型一氧化氮合酶（iNOS）。
结果
牵张显著诱导NRK-52E细胞凋亡，同时伴有TGF-β释放增加；1D11（10μg/mL）完全抑制牵张诱导的凋亡。与未梗阻肾脏相比，对照梗阻肾脏中的TGF-β含量高20倍；1D11中和了梗阻肾脏中的组织TGF-β。与未梗阻的对应肾脏相比，对照梗阻肾脏表现出明显更多的纤维化和肾小管凋亡，而1D11可减轻这种情况。相反，1D11显著增加肾小管增殖。p53免疫染色定位于对照梗阻肾脏的肾小管细胞核，而1D11使其减少。相反，在1D11处理的梗阻肾脏中bcl-2上调。1D11处理增加了梗阻肾脏的总NOS活性和iNOS活性。
结论
本研究强烈表明，抗TGF-β抗体是预防UUO中肾小管纤维化和凋亡的有前景的药物。
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