Avastin对翼状胬肉成纤维细胞增殖的影响

发布时间：2018-07-21 19:55

【摘要】：目的评价Avastin(贝伐单抗)在体外对翼状胬肉成纤维细胞的增殖抑制作用以及细胞毒性。方法通过不同浓度Avastin(0.25g·L-1、0.50g·L-1、1.00g·L-1、1.50g·L-1、2.50g·L-1)处理人翼状胬肉成纤维细胞12h、24h、48h、72h,观察成纤维细胞的增殖抑制、药物细胞毒性以及凋亡蛋白的表达。WST-1试剂检测细胞增殖,LDH试剂盒检测细胞毒性,Western blot方法检测增殖蛋白以及凋亡蛋白的表达。结果 48h及72hAvastin组与正常组相比光密度(OD)值差异有统计学意义(P=0.034、P=0.016)。与12h相比,0.25g·L-1 Avastin组及0.50g·L-1 Avastin组作用24h、48h、72h后细胞增殖抑制率均无明显提高(均为P0.05),1.50g·L-1 Avastin组及2.50g·L-1 Avastin组作用48h、72h后细胞增殖抑制率均明显提高(均为P0.05)。与0.25g·L-1 Avastin组相比,0.50g·L-1 Avastin组在各个时间点(除12h外)细胞增殖抑制率均无明显提高(均为P0.05),1.50g·L-1 Avastin组、2.50g·L-1 Avastin组在各个时间点抑制率均明显提高(均为P0.05)。Avastin各浓度组与LDH最大释放组(加入裂解酶)对翼状胬肉成纤维细胞LDH释放率差异均有统计学意义(均为P0.05),各浓度组之间差异均无统计学意义(均为P0.05)。Avastin可抑制p-CyclinD1表达,促进Caspase3的表达,与细胞对照组相比差异均具有统计学意义(均为P0.05)。结论 Avastin可以通过抑制p-CyclinD1以及促进Caspase3的表达抑制翼状胬肉成纤维细胞的增殖,但无细胞毒性,进一步证实了该药物的安全有效性。
[Abstract]:Objective to evaluate the proliferation inhibition and cytotoxicity of Avastin on pterygium fibroblasts in vitro. Methods Human pterygium fibroblasts were treated with different concentrations of Avastin (0.25g / L ~ (-1) 0.50g / L ~ (-1) ~ 1.50g / L ~ (-1) for 12 h ~ (24) h ~ (48) h ~ (72) h, and the proliferation inhibition of human pterygium fibroblasts was observed. Cytotoxicity and expression of apoptotic protein. WST-1 reagent was used to detect the expression of proliferating protein and apoptotic protein by Western blot. Results there was significant difference in optical density (OD) between 48 h and 72hAvastin group and normal group (P < 0.034). Compared with 12h, the inhibition rate of cell proliferation in 0.25 g L-1 Avastin group and 0.50 g L-1 Avastin group was not significantly increased (P0.05) after treatment for 24 h or 48 h or 72 h (P0.05) in 1.50 g L-1 Avastin group and 2.50 g L-1 Avastin group at 48 h and 72 h after treatment (P0.05). Compared with 0.25 g L-1 Avastin group, 0.50 g L-1 Avastin group had no significant increase in cell proliferation inhibition rate at all time points (except 12 h) (P0.05). The inhibitory rate of 1.50 g L-1 Avastin group was significantly higher than that of 0.25 g L-1 Avastin group at each time point (all P0.05). Avastin concentration group and maximum release rate of Avastin group were significantly higher than that of 0.25 g L-1 Avastin group. The LDH release rate of pterygium fibroblasts was significantly different (P0.05), but there was no significant difference (P0.05). Avastin could inhibit the expression of p-CyclinD1 in pterygium fibroblasts. Promote Caspase3 expression, compared with the cell control group, there was significant difference (P0.05). Conclusion Avastin can inhibit the proliferation of pterygium fibroblasts by inhibiting the expression of p-CyclinD1 and Caspase3, but has no cytotoxicity.
【作者单位】：广州医科大学附属第二医院眼科教研室;
【基金】：广东省科技计划项目基金资助(编号:20100309)~~
【分类号】：R777.33

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