Important metabolites in maintaining folic acid, homocysteine and polyamine metabolism associated with ranibizumab treatment in cultured Human Tenon’s Fibroblast
Anti-fibrotic properties of ranibizumab have been well documented. As an antagonist to Vascular Endothelial Growth Factor (VEGF), ranibizumab works by binding and neutralizes all active VEGF, thus limits the progressive cell growth and proliferation. Its application in ocular diseases has shown remarkable desired effects, however to date its anti-fibrotic mechanism is not well understood. In this study, we identified metabolic changes in ranibizumab-treated Human Tenos’s fibroblast (HTF). Cultured HTFs were treated for48 hours with 0.5 mg/ml of ranibizumab and 0.5 mg/ml control IgG antibody which serves as a negative control. Samples from each group were injected into Agilent 6520 Q-TOF Liquid Chromatography/ Mass Spectrometer (LC/MS) System to establish the metabolite expression in both ranibizumab treated cells and control group. Data obtained was analysed using Agilent Mass Hunter Qualitative Analysis software to identify the most regulated metabolite following ranibizumab treatment. At statistical analysis of p-value < 0.01 with the cut off value of two-fold change, 31 identified metabolites were found to be significantly up-regulated in ranibizumab-treated group, with 6 of the mostly up-regulated have insignificant role in fibroblast’s cell cycle and wound healing regulations Meanwhile, 121 identified metabolites were down-regulated with seven of the mostly down-regulated are significantly involved in cell cycle and proliferation. Our findings demonstrated that ranibizumab abrogates the tissue scarring process and wound healing formation by regulating the expression of metabolites associated with fibrotic activity. In particular, we found that vitamin Bs are important in maintaining normal folic acid cycle, nucleotide synthesis, homocysteine and spermidine metabolism. This study provides an insight of ranibizumab mechanism of actions on HTFs from the perspective of metabolomics.