研究目的
To investigate the effects of anti-vascular endothelial growth factor (VEGF) antibody on the survival of retinal ganglion cell (RGC)-5 cells differentiated with staurosporine under oxidative stress.
研究成果
Bevacizumab administration increases cell damage in differentiated RGC-5 cells under oxidative stress, with cytotoxicity rising with higher concentrations. VEGF, VEGFR-1, and VEGFR-2 expressions were upregulated with bevacizumab treatment. Caution is advised in clinical use of bevacizumab, especially in conditions like glaucoma, and further in vivo studies are needed.
研究不足
The study was conducted in vitro using the RGC-5 cell line, which may not fully represent in vivo conditions or all retinal ganglion cells. The RGC-5 cell line has been reported to be cross-contaminated with a mouse fibroblast cell line, potentially affecting results. No in vivo experiments were performed, limiting the understanding of interactions with other cells and mechanisms in the human body.
1：Experimental Design and Method Selection:
The study used in vitro cell culture experiments with RGC-5 cells differentiated using staurosporine. Methods included real-time PCR and Western blot for gene and protein expression analysis, and lactate dehydrogenase (LDH) assay for cell survival assessment under oxidative stress induced by hydrogen peroxide (H2O2).
2：2). Sample Selection and Data Sources:
2. Sample Selection and Data Sources: The RGC-5 cell line was used, which is an immortalized mouse retinal ganglion cell line. Cells were differentiated with 1 μg staurosporine for 6 hours.
3：List of Experimental Equipment and Materials:
Equipment and materials included staurosporine (Sigma), H2O2, bevacizumab (Avastin; Genentech), real-time PCR kits (QuantiTect SYBR Green PCR kit, Qiagen), Western blot reagents (e.g., Immobilon-P transfer membrane, Millipore), LDH assay kit (Calbiochem-Novabiochem), and statistical software (SPSS ver.
4：0). Experimental Procedures and Operational Workflow:
Cells were differentiated, treated with H2O2 and bevacizumab at various concentrations, and then analyzed for survival via LDH assay and for expression of VEGF, VEGFR-1, and VEGFR-2 via real-time PCR and Western blot.
5：Data Analysis Methods:
Data were analyzed using the Mann-Whitney U-test in SPSS, with p < 0.05 considered significant.
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