Department of Ophthalmology, Shanghai Fourth People's Hospital, Tongji University School of Medicine, No. 1279, Sanmen road, Shanghai, 200434, China.
Department of Ophthalmology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, No. 85 / 86, Wujin road, Shanghai, 200080, China.
BMC Ophthalmol. 2023 Apr 26;23(1):183. doi: 10.1186/s12886-023-02914-4.
Safely inhibiting the formation of scar in the glaucoma filtration surgery (GFS) has always been an issue for clinical glaucoma doctors. Anti-vascular endothelial growth factor (VEGF) agents can reduce angiogenesis, and anti-placental growth factor (PIGF) agents can affect reactive gliosis. However, the effect of conbercept, which can bind to both VEGF and PIGF, on human Tenon's fibroblasts (HTFs) is unknown.
HTFs were cultured in vitro and treated with conbercept or bevacizumab (BVZ). No drug was added to the control group. The effects of drugs on cell proliferation were assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and the collagen type I alpha1(Col1A1) mRNA expression level was measured using quantitative polymerase chain reaction (qPCR). HTF cell migration after drug interventions was evaluated using the scratch wound assay along with the measurement of the expression levels of VEGF and PIGF in human umbilical vein endothelial cells (HUVECs) using enzyme-linked immunosorbent assay, as well as the detection of the VEGF(R) mRNA expression level in HTFs using qPCR.
After the addition of conbercept (0.01, 0.1, and 1 mg/mL) to the cultured HTFs or HUVECs, no significant cytotoxicity was observed compared with the control group, while the cytotoxicity of 2.5 mg/mL BVZ on HTFs was obvious. Conbercept significantly inhibited HTF cell migration and Col1A1 mRNA expression level in HTFs. It was superior to BVZ in inhibiting HTF migration. After the intervention with conbercept, the expression level of PIGF and VEGF in HUVECs significantly decreased; and the inhibitory effect of conbercept on the expression level of VEGF in HUVECs was weaker than that of BVZ. Conbercept was more advantageous than BVZ in inhibiting the expression level of VEGFR-1 mRNA in HTFs. However, its effect in terms of inhibiting the expression level of VEGFR-2 mRNA in HTFs was weaker than that of BVZ.
The results suggested the low cytotoxicity and significant anti-scarring effect of conbercept in HTF with significant anti-PIGF and inferior anti-VEGF effects compared with BVZ, thus providing a better understanding of the role of conbercept in the GFS wound healing process.
安全抑制青光眼滤过手术(GFS)中的瘢痕形成一直是临床青光眼医生关注的问题。抗血管内皮生长因子(VEGF)药物可以减少血管生成,抗胎盘生长因子(PIGF)药物可以影响反应性神经胶质增生。然而,康柏西普(conbercept)既能结合 VEGF 又能结合 PIGF,对人眼Tenon's 纤维细胞(HTFs)的作用尚不清楚。
体外培养 HTFs,并用康柏西普或贝伐珠单抗(BVZ)处理。对照组不加药物。用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)法评估药物对细胞增殖的影响,用定量聚合酶链反应(qPCR)法测量胶原 I alpha1(Col1A1)mRNA 表达水平。用划痕实验评估药物干预后 HTF 细胞迁移情况,并通过酶联免疫吸附试验(ELISA)检测人脐静脉内皮细胞(HUVECs)中 VEGF 和 PIGF 的表达水平,以及 qPCR 法检测 HTFs 中 VEGF(R)mRNA 表达水平。
与对照组相比,加入浓度为 0.01、0.1 和 1mg/ml 的康柏西普到培养的 HTFs 或 HUVECs 中,未见明显细胞毒性,而 2.5mg/mlBVZ 对 HTFs 的细胞毒性明显。康柏西普显著抑制 HTF 细胞迁移和 HTFs 中 Col1A1mRNA 表达水平,在抑制 HTF 迁移方面优于 BVZ。康柏西普干预后,HUVECs 中 PIGF 和 VEGF 的表达水平显著下降,康柏西普对 HUVECs 中 VEGF 表达水平的抑制作用弱于 BVZ。康柏西普在抑制 HTFs 中 VEGFR-1mRNA 表达水平方面优于 BVZ,但抑制 HTFs 中 VEGFR-2mRNA 表达水平的作用弱于 BVZ。
结果提示康柏西普具有低细胞毒性和显著的抗瘢痕形成作用,在抑制 PIGF 方面作用显著,在抑制 VEGF 方面作用弱于 BVZ,为了解康柏西普在 GFS 伤口愈合过程中的作用提供了更好的认识。