Liang Ze-Yu, Wang Yi-Peng, Li Jing, Yang Wen-Chao, Tu Yong-Fang, Zhang Yue, Chen Song
Tianjin Eye Hospital, Tianjin Eye Institute, Tianjin Key Lab of Ophthalmology and Visual Science, Nankai University Affiliated Eye Hospital, Tianjin 300020, China.
Anyang Eye Hospital, Anyang 455000, Henan Province, China.
Int J Ophthalmol. 2022 Oct 18;15(10):1577-1585. doi: 10.18240/ijo.2022.10.03. eCollection 2022.
To assess the effects of intravitreal ranibizumab (IVR) on angiogenesis and glial activity of the fibrovascular membrane (FVM) in patients with proliferative diabetic retinopathy (PDR).
Forty-two eyes from 42 patients with PDR requiring vitrectomy were included and divided into two groups: control group (=16) did not receive IVR, while IVR group (=26) underwent IVR 5d before vitrectomy. FVM specimens were collected by the same surgeon during the interventions. Histopathological morphology was examined by hematoxylin-eosin (H-E) staining and cell densities in the FVM was assessed. Microvessels were outlined by immunohistochemical staining of CD31 and microvessel density (MVD) assessed as an index of FVM angiogenesis. Dual-color immunofluorescence staining, and confocal microscopy was used to detect co-localization and relative expression levels of glial fibrillary acidic protein (GFAP) and α-smooth muscle actin (α-SMA) as markers of glial-mesenchymal transition (GMT). The GMT index (GI; ratio of relative GFAP/α-SMA expression) was used to semi-quantify the degree of GMT or glial activity of FVMs.
H-E staining showed similar vascularization in both groups, with microvessels and scattered stromal cells in the matrix. Infiltrated cell densities did not differ significantly between the two groups (>0.05). The MVD of the IVR group (130.62±15.46/mm) was significantly lower than that of the controls (142.25±19.16/mm, <0.05). In both groups, all sections were strongly immunostained for GFAP and α-SMA. The Pearson's correlation coefficients (PCC) of intensity of automated pixel count of two markers indicated GFAP and α-SMA co-stained well and GMT participated in the remolding of FVMs in PDR. The mean relative GFAP expression in the IVR group was significantly lower, whereas that of α-SMA was significantly higher than in controls (<0.05). GI in the IVR group (1.10±0.10) was significantly lower than in the controls (1.21±0.12, <0.05).
IVR can reduce angiogenesis, glial activity of FVM and promote glial-fibrotic transformation by reducing MVD and promoting GMT but does not decrease the cell density in patients with PDR.
评估玻璃体内注射雷珠单抗（IVR）对增殖性糖尿病视网膜病变（PDR）患者纤维血管膜（FVM）血管生成和神经胶质细胞活性的影响。
纳入42例需要进行玻璃体切除术的PDR患者的42只眼，分为两组：对照组（n = 16）未接受IVR，而IVR组（n = 26）在玻璃体切除术前5天接受IVR。在干预过程中由同一位外科医生收集FVM标本。通过苏木精 - 伊红（H-E）染色检查组织病理学形态，并评估FVM中的细胞密度。通过CD31免疫组织化学染色勾勒微血管，并评估微血管密度（MVD）作为FVM血管生成的指标。采用双色免疫荧光染色和共聚焦显微镜检测神经胶质纤维酸性蛋白（GFAP）和α-平滑肌肌动蛋白（α-SMA）的共定位及相对表达水平，作为神经胶质 - 间充质转化（GMT）的标志物。GMT指数（GI；相对GFAP/α-SMA表达的比值）用于半定量FVM的GMT程度或神经胶质细胞活性。
H-E染色显示两组的血管化情况相似，基质中有微血管和散在的基质细胞。两组间浸润细胞密度无显著差异（P>0.05）。IVR组的MVD（130.62±15.46/mm）显著低于对照组（142.25±19.16/mm，P<0.05）。在两组中，所有切片对GFAP和α-SMA均有强免疫染色。两种标志物自动像素计数强度的Pearson相关系数（PCC）表明GFAP和α-SMA共染色良好，且GMT参与了PDR中FVM的重塑。IVR组中GFAP的平均相对表达显著较低，而α-SMA的平均相对表达显著高于对照组（P<0.05）。IVR组的GI（1.10±0.10）显著低于对照组（1.21±0.12，P<0.05）。
IVR可通过降低MVD和促进GMT来减少PDR患者FVM的血管生成、神经胶质细胞活性，并促进神经胶质 - 纤维化转化，但不降低细胞密度。
