Protective effects on the retina after ranibizumab treatment in an ischemia model
Protective effects on the retina after ranibizumab treatment in an ischemia model
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DOI：
10.1371/journal.pone.0182407
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发表时间：
2017-08-11
期刊：
影响因子：
3.7
通讯作者：
Dick, H. Burkhard
中科院分区：
文献类型：
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作者：
Joachim, Stephanie C.;Renner, Marina;Dick, H. Burkhard
视网膜缺血在糖尿病视网膜病变或视网膜血管阻塞等眼科疾病中很常见。本研究的目的是通过免疫组织化学(IF)和实时定量聚合酶链式反应(PCR)来评价玻璃体内注射血管内皮生长因子(VEGF)抑制剂(Ranibizumab)对缺血动物模型视网膜细胞的潜在保护作用。斑点杂交证实雷尼比珠单抗与大鼠血管内皮细胞生长因子-A呈阳性结合。一只眼发生缺血，另一组接受雷尼比珠单抗治疗。缺血眼房水中血管内皮生长因子水平显著升高(p=0.032)，而雷尼比珠单抗眼房水中血管内皮生长因子水平较低(p=0.99)。缺血视网膜表现为视网膜神经节细胞数显著减少(RGC；如果BRN-3a：P<；0.001，如果RBPMS：P<；0.001；PCr：P=0.002)。雷尼比珠单抗组RGC明显少于缺血组(Brn-3a：P=0.001；P=0.007)，但多于缺血组。缺血后光感受器面积减少(IF：P=0.049；PCRP=0.511)，而雷尼比珠单抗组(IF：P=0.947；PCRP=0.122)与对照组无明显差异。在缺血组(p<；0.001)和雷尼比珠单抗组(p<；0.001)，ChAT(+)无长突细胞减少，而雷尼比珠单抗组减少较少。血管内皮生长因子受体2(VEGFR2；IF：P<；0.001；聚合酶链式反应：P=0.021)和大胶质细胞(GFAP；IF：P<；0.001；聚合酶链式反应：P<；0.001)活化。在雷尼比珠单抗视网膜中激活较弱(血管内皮生长因子-R2：IF：P=0.1；聚合酶链式反应：P=0.03；胶质纤维酸性蛋白：如果：P=0.1；聚合酶链式反应：P=0.015)。脑缺血后小胶质细胞总数(IF：P=0.003；P=0.023；P=0.023)和活化小胶质细胞(IF：P<；0.001；P=0.009)数量明显增加。雷尼比珠单抗组的上述水平较高(IBA1：IF：P<；0.001；PCr：P=0.018；CD68：IF：P<；0.001；Pcr：P=0.004)。我们的发现表明，雷尼比珠单抗治疗可以保护光感受器和视网膜节细胞。只有无长突细胞是无法挽救的。他们似乎对缺血损伤特别敏感，可能需要更早的干预。
Retinal ischemia is common in eye disorders, like diabetic retinopathy or retinal vascular occlusion. The goal of this study was to evaluate the potential protective effects of an intravitreally injected vascular endothelial growth factor (VEGF) inhibitor (ranibizumab) on retinal cells in an ischemia animal model via immunohistochemistry (IF) and quantitative real-time PCR (PCR). A positive binding of ranibizumab to rat VEGF-A was confirmed via dot blot. One eye underwent ischemia and a subgroup received ranibizumab. A significant VEGF increase was detected in aqueous humor of ischemic eyes (p = 0.032), whereas VEGF levels were low in ranibizumab eyes (p = 0.99). Ischemic retinas showed a significantly lower retinal ganglion cell number (RGC; IF Brn-3a: p < 0.001, IF RBPMS: p < 0.001; PCR: p = 0.002). The ranibizumab group displayed fewer RGCs (IF Brn-3a: 0.3, IF RBPMS: p < 0.001; PCR: p = 0.007), but more than the ischemia group (IF Brn-3a: p = 0.04, IF RBPMS: p = 0.03). Photoreceptor area was decreased after ischemia (IF: p = 0.049; PCR: p = 0.511), while the ranibizumab group (IF: p = 0.947; PCR: p = 0.122) was comparable to controls. In the ischemia (p < 0.001) and ranibizumab group (p < 0.001) a decrease of ChAT(+) amacrine cells was found, which was less prominent in the ranibizumab group. VEGF-receptor 2 (VEGF-R2; IF: p < 0.001; PCR: p = 0.021) and macroglia (GFAP; IF: p < 0.001; PCR: p < 0.001) activation was present in ischemic retinas. The activation was weaker in ranibizumab retinas (VEGF-R2: IF: p = 0.1; PCR: p = 0.03; GFAP: IF: p = 0.1; PCR: p = 0.015). An increase in the number of total (IF: p = 0.003; PCR: p = 0.023) and activated microglia (IF: p < 0.001; PCR: p = 0.009) was detected after ischemia. These levels were higher in the ranibizumab group (Iba1: IF: p < 0.001; PCR: p = 0.018; CD68: IF: p < 0.001; PCR: p = 0.004). Our findings demonstrate that photoreceptors and RGCs are protected by ranibizumab treatment. Only amacrine cells cannot be rescued. They seem to be particularly sensitive to ischemic damage and need maybe an earlier intervention.