Effect of Mingjing Granules on Fibrovascular Membrane of Wet Age-related Macular Degeneration Based on Macrophages and Glial Cells

LI Xiaoyu; LIANG Lina; GAO Yun

doi:10.13422/j.cnki.syfjx.20231015

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Effect of Mingjing Granules on Fibrovascular Membrane of Wet Age-related Macular Degeneration Based on Macrophages and Glial Cells

更新时间：2023-10-20

- Effect of Mingjing Granules on Fibrovascular Membrane of Wet Age-related Macular Degeneration Based on Macrophages and Glial Cells
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 29, Issue 22, Pages: 95-103(2023)
- 作者机构：
  
  中国中医科学院眼科医院，北京 100040
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20231015
  CLC： R284;R285;R289;R287;R22;R2-031;R33;R24
- Published：20 November 2023，
  
  Published Online：18 August 2023，
  
  Received：14 April 2023，
- 稿件说明：
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李晓宇,梁丽娜,高云.基于巨噬细胞和胶质细胞探讨明睛颗粒对湿性年龄相关性黄斑变性纤维血管膜的影响[J].中国实验方剂学杂志,2023,29(22):95-103.

LI Xiaoyu,LIANG Lina,GAO Yun.Effect of Mingjing Granules on Fibrovascular Membrane of Wet Age-related Macular Degeneration Based on Macrophages and Glial Cells[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(22):95-103.
李晓宇,梁丽娜,高云.基于巨噬细胞和胶质细胞探讨明睛颗粒对湿性年龄相关性黄斑变性纤维血管膜的影响[J].中国实验方剂学杂志,2023,29(22):95-103. DOI： 10.13422/j.cnki.syfjx.20231015.

LI Xiaoyu,LIANG Lina,GAO Yun.Effect of Mingjing Granules on Fibrovascular Membrane of Wet Age-related Macular Degeneration Based on Macrophages and Glial Cells[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(22):95-103. DOI： 10.13422/j.cnki.syfjx.20231015.

摘要

目的

基于巨噬细胞和胶质细胞探讨明睛颗粒（MJKL）对实验性湿性年龄相关性黄斑变性（nAMD）纤维血管膜的影响，进一步阐释MJKL治疗nAMD的作用机制。

方法

通过两阶段激光光凝建立实验性nAMD纤维血管膜模型，将造模成功后的BN大鼠随机分为模型组、抗血管内皮生长因子（VEGF）组（5 μL/眼）、MJKL+抗VEGF组（1 g·mL

-1

+5 μL/眼）3组。模型组，给予蒸馏水灌胃；抗VEGF组，予玻璃体腔注射雷珠单抗注射液；MJKL+抗VEGF组，予玻璃体腔注射雷珠单抗注射液，同时开始予MJKL灌胃。10只正常BN大鼠不造模，常规饲养作为正常组。造模40 d后，采用眼底照相（FP）、眼底血管荧光造影（FFA）、光学相干断层扫描（OCT）、视网膜色素上皮（RPE）-脉络膜-巩膜铺片观察眼底病变形态、病变渗出面积及吸光度

；组织病理学观察视网膜结构的改变，免疫荧光法检测F4/80、离子钙接头蛋白抗原（Iba-1）、胶质纤维酸性蛋白（GFAP）的表达及分布，实时荧光定量聚合酶链反应（Reat-time PCR）检测F4/80、Iba-1、GFAP mRNA的相对表达量。

结果

两阶段激光造模后40 d建立纤维血管膜模型；与模型组比较，抗VEGF组在病变渗出面积、

、病变高度、病变面积均明显降低（

0.05），视网膜结构损伤程度显著改善；而MJKL+抗VEGF组较抗VEGF组在渗出面积、

、病变高度、病变面积明显降低（

0.05），病变面积及视网膜结构损伤程度改善更为明显。与正常组比较，模型组F4/80与Iba-1的荧光强度均明显升高（

0.05）；与模型组比较，抗VEGF组F4/80与Iba-1的荧光强度均明显降低（

0.05）；与抗VEGF组比较，MJKL+抗VEGF组F4/80与Iba-1的荧光强度明显降低（

0.05）。与正常组比较，模型组GFAP的荧光强度明显升高（

0.05）；与模型组比较，抗VEGF组GFAP的荧光强度明显降低（

0.05）。与正常组比较，模型组F4/80、Iba-1、GFAP mRNA的相对表达量均明显升高（

0.05）；与模型组比较，抗VEGF组F4/80、Iba-1、GFAP mRNA的相对表达量均明显降低（

0.05）；与抗VEGF组比较，MJKL+抗VEGF组F4/80、Iba-1、GFAP mRNA的相对表达量均明显降低（

0.05）。

结论

MJKL联合抗VEGF药物可以较单一应用抗VEGF药物更好地抑制实验性nAMD纤维血管膜的生长，其机制可能与抑制巨噬细胞与胶质细胞参与血管膜形成有关。

Abstract

Objective

To investigate the effects of Mingjing granules （MJKL） on the fibrovascular membrane of experimental wet age-related macular degeneration （nAMD） based on macrophages and glial cells and further explain the mechanism of MJKL in the treatment of nAMD.

Method

The experimental nAMD fibrovascular membrane model was established by two-stage laser photocoagulation. BN rats were randomly divided into three groups： model group， anti-vascular endothelial growth factor （VEGF） group， and MJKL + anti-VEGF group. The model group was given distilled water for intragastric administration. Anti-VEGF group was injected with leizumab injection in the vitreous cavity. MJKL + anti-VEGF group was injected with leizumab injection in the vitreous cavity， and MJKL was intragastrically administered. Ten normal BN rats were not modeled and fed as controls. After 40 days of model making， fundus lesion morphology， lesion exudation area， and MD value were observed by fundus photography （FP）， fundus angiography （FFA）， optical coherence tomography （OCT）， and retinal pigment epithelium （RPE）-choroid-sclera film. The changes in retinal structure were observed by histopathology， and the expression and distribution of F4/80， Iba-1， and GFAP were detected by immunofluorescence. The relative expression levels of F4/80， Iba-1， and GFAP mRNA were detected by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）.

Result

The fibrovascular membrane model was established 40d after two-stage laser modeling. The lesion exudation area， MD value， lesion height， and lesion area in the anti-VEGF group were significantly lower than those in the model group （

0.05）， and the retinal structural damage degree was significantly improved. Compared with the anti-VEGF group， the MJKL + anti-VEGF group significantly decreased the MD value， lesion height， and lesion area （

0.05）， and lesion area and retinal structural damage degree were significantly improved. The fluorescence intensity of F4/80 and Iba-1 in the model group was significantly higher than that in the normal group （

0.05）， and that in the anti-VEGF group was significantly lower than that in the model group （

0.05）. The fluorescence intensity in the MJKL + anti-VEGF group was significantly lower than that in the anti-VEGF group （

0.05）. The fluorescence intensity of GFAP in the model group was significantly higher than that in the normal group （

0.05）， and that in the anti-VEGF group was significantly lower than that in the model group （

0.05）. The relative expression levels of F4/80， Iba-1， and GFAP mRNA in the model group were significantly increased compared with the normal group （

0.05）， and the anti-VEGF group was significantly decreased compared with the model group （

0.05）. The relative expression levels of F4/80， Iba-1， and GFAP mRNA in the MJKL + anti-VEGF group were significantly decreased compared with those in the anti-VEGF group （

0.05）.

Conclusion

MJKL combined with anti-VEGF drugs can inhibit the growth of experimental nAMD fibrovascular membrane better than anti-VEGF drugs alone， and the mechanism may be related to inhibiting the participation of macrophages and glial cells in the formation of fibrovascular membrane.

关键词

明睛颗粒两阶段激光诱导纤维血管膜巨噬细胞胶质细胞

Keywords

Mingjing granulestwo-stage laser inductionfibrovascular membranemacrophageglial cell

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