Antibody to transforming growth factor-β ameliorates tubular apoptosis in unilateral ureteral obstruction
Antibody to transforming growth factor-β ameliorates tubular apoptosis in unilateral ureteral obstruction
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DOI：
10.1046/j.1523-1755.2000.00414.x
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发表时间：
2000-12-01
影响因子：
19.6
通讯作者：
Felsen, D
中科院分区：
文献类型：
--
作者：
Miyajima, A;Chen, J;Felsen, D
背景。单侧输尿管梗阻（UUO）以进行性肾萎缩、肾间质纤维化、肾转化生长因子- β (tgf - β)升高和肾小管凋亡为特征。本研究旨在确定tgf - β单克隆抗体（ID11）在uu中的作用。通过计算机辅助系统对小管上皮细胞（NRK-52E）施加机械拉伸。在UUO前1天和之后每2天给药3次（0.5、2或4 mg/大鼠），并在第13天取肾。通过测定组织羟脯氨酸和胶原、纤维连接蛋白mRNA来评估纤维化程度。用末端脱氧转移酶尿苷三磷酸缺口末端标记法评估细胞凋亡。采用生物测定法测定tgf - β水平。Western blot和免疫染色检测增殖细胞核抗原（PCNA）、p53、bcl-2和诱导型一氧化氮合酶（iNOS）。拉伸显著诱导NRK-52E细胞凋亡，并伴有tgf - β释放增加；1D11（10杯/mL）完全抑制拉伸诱导的细胞凋亡。对照组梗阻肾的tgf - β含量比未梗阻肾高20倍；1D11能中和阻塞肾脏的组织tgf - β。对照组梗阻肾比未梗阻肾表现出更多的纤维化和小管凋亡，而未梗阻肾则被1D11钝化。相反，1D11显著增加小管增殖。p53的免疫染色定位于对照组梗阻肾小管核，并被1D11减弱。相比之下，bcl-2在1d11治疗的梗阻肾脏中上调。1D11治疗可提高梗阻肾总NOS活性和iNOS活性。本研究强烈提示，tgf - β抗体是预防UUO肾小管纤维化和凋亡的有希望的药物。
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 (ID11) 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 mug/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.