Application of anti-adhesion molecule therapy for glomerulonephritis -Effects of sulfated oligosaccharides as selectin-blocking agents-

背景资料。选择素是一种黏附分子，可介导白细胞向炎症组织的渗透。P-选择素和L-选择素与配体分子上的硫酸化低聚糖链结合。人工合成的硫酸化低聚糖，包括硫酸化透明质酸和硫酸胶原酸，在体外均能抑制P选择素和L选择素依赖的黏附通路。我们评价了SHA和SCA对大鼠系膜增生性肾炎和大鼠新月体肾炎的预防作用。雌性Wistar大鼠单侧肾切除后注射抗Thy-1抗体建立系膜增生性肾炎模型。雌性WKY大鼠注射肾毒性血清复制新月体肾炎模型。用抗大鼠P-选择素和L-选择素、透明质酸、雪花酸和可可酸的中和性和非中和性单抗处理大鼠。免疫组织化学方法检测P-选择素和巨噬细胞的定位。实时荧光定量RT-PCR法检测肾小球PDGF B链基因表达。两种模型肾小球P-选择素表达增强，巨噬细胞浸润明显。抗P-选择素单抗能显著减少蛋白尿、新月体形成和巨噬细胞的肾小球浸润，而抗L-SL单抗则无明显作用。SHA和SCA以剂量依赖方式减少蛋白尿、巨噬细胞浸润和新月体形成。SHA组和SCA组PDGF B链基因表达明显降低。综上所述，SHA和SCA通过阻断P-选择素依赖的黏附途径抑制肾小球巨噬细胞的浸润，阻止系膜增生性肾小球肾炎和大鼠新月体肾炎的进展。我们的结论是，硫酸寡糖可能对治疗肾小球肾炎有好处。

Background. Selectins are adhesion molecules which mediate leukocyte infiltration into inflammatory tissues. P- and L-selectin bind to sulfated oligosaccharide chains on the ligand molecules. Synthesized sulfated oligosaccharides, including sulfated hyarulonic acid (SHA) and sulfated colominic acid (SCA), inhibit both P- and L-selectin-dependent adhesion pathways in vitro. We evaluated the preventive effects of SHA and SCA on rat mesangial proliferative glomerulonephritis and rat crescentic glomerulonephritis.Methods. Female Wistar rats were injected with anti-Thy-1 antibody after unilateral nephrectomy for a mesangial proliferative glomerulonephritis model. Female WKY rats were injected with nephrotoxic serum for a crescentic glomerulonephritis model. Rats were administered with neutralizing or non-neutralizing monoclonal antibodies to rat P-selectin and L-selectin, SHA, hyarulonic acid, SCA and colominic acid. Localization of P-selectin and macrophages were examined by immunohistochemical methods. Gene expression of PDGF B chain in the glomeruli was quantified using real-time RT-PCR method.Results. Increased expression of P-selectin and macrophage infiltration was prominent in the glomeruli in both two models. Proteinuria, crescent formation and glomerular infiltration of macrophages were significantly reduced by anti-P-selectin mAb, but not by anti-L-SL mAb. SHA and SCA reduced proteinuria, macrophage infiltration and crescent formation in dose dependent manner. Gene expression of PDGF B chain was significantly decreased in SHA and SCA treatment groups. In conclusion, SHA and SCA inhibited glomerular macrophage infiltration by blocking P-selectin-dependent adhesion pathway, and prevented disease progression in rat mesangial proliferative glomerulonephritris and rat crescentic glomerulonephritis. We conclude that sulfated oligosaccharides may be beneficial for the treatment of glomerulonephritis.