| 加减驻景方含药血清对AKT基因转染后ARPE-19细胞表达VEGF的影响 |
| 投稿时间：2018-02-24 修订日期：2018-04-01 点此下载全文 |
| 引用本文：褚文丽,陈水龄,亢泽峰,张丛青,李维义,刘健.加减驻景方含药血清对AKT基因转染后ARPE-19细胞表达VEGF的影响[J].医学研究杂志,2018,47(12):55-58 |
| DOI：
10.11969/j.issn.1673-548X.2018.12.014 |
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| 基金项目:国家自然科学基金资助项目（面上项目）（81574032） |
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| 中文摘要:目的 探讨加减驻景方含药血清对AKT基因转染后人视网膜色素上皮细胞（ARPE-19）细胞表达VEGF的影响。方法 制备SD大鼠加减驻景方含药血清及空白血清，建立AKT基因转染ARPE-19细胞模型，体外培养，分为5个组，正常组为ARPE-19细胞常规培养，转染模型组（模型细胞，常规条件培养），含药血清组（模型细胞，给予含药血清干预培养）、空白血清组（模型细胞，给予空白血清干预培养）和阳性对照组（模型细胞，给予康柏西普干预培养）。设立不同时间和不同浓度，通过CCK-8法检测各组血清及阳性组药物对细胞毒性、增殖的影响；通过ELISA检测各组细胞上清液中VEGF的表达。结果 蛋白质印迹法（Western blot）显示，转染模型组与正常组比较，AKT蛋白的表达量明显增加，差异有统计学意义（P<0.05）。CCK-8示，细胞培养24、48h后，AKT基因转染可促进ARPE-19细胞增殖（P<0.05），培养72h后，对细胞增殖影响不大（P > 0.05）。10、20μg/ml康柏西普及5%、10%的含药血清对模型细胞的增殖有抑制作用（P<0.05），各浓度空白血清组对模型细胞组的增殖影响不大（P > 0.05）。酶联免疫吸附试验（ELISA）检测显示：转染模型组细胞上清液中VEGF表达量高于正常组（P<0.05），康柏西普组与含药血清组均显著低于模型组（P<0.05），其中康柏西普组低于含药血清组，差异有统计学意义（P<0.05）。空白血清组与模型组比较，差异无统计学意义（P > 0.05）。结论 AKT基因转染后可诱导ARPE-19细胞的增殖，促进VEGF的表达。加减驻景方含药血清对AKT基因转染后ARPE-19细胞的增殖及VEGF的表达有抑制作用。加减驻景方可能通过抑制VEGF的过度表达发挥干预CNV生成的作用。 |
| 中文关键词:加减驻景方 含药血清 ARPE-19细胞 基因转染 血管内皮生长因子 |
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| Effects of Jiajianzhujing Decoction Containing Serum on Expression of VEGF of ARPE-19 Cells Transfected with AKT Gene |
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| Abstract:Objective To investigate the effect of addition and subtraction of sera containing medicinal herbs on the expression of VEGF in human retinal pigment epithelial cells (ARPE-19) cells after AKT gene transfection. Methods SD rats serum containing the prescription of Jiajianzhujing and the content of distilled water in serum were prepared.AKT gene transfected ARPE-19 cell model was established and Western blot method was used to verify the successful construction. The effects of the prescription and distilled water in serum at different concentration (2.5%, 5%, 10%) and Conbercept (5, 10, 20g/ml) cell vitality was observed by cell counting kit(CCK-8) assay. The effect of serum and Conbercept on the expression of VEGF in model cells was assessed by ELISA. Results Result of Western blot assay confirmed that:compared with the normal group, the expression of AKT protein increased significantly in the model group(P<0.05). Results of CCK-8 measure confirmed that After be cultured for 24h and 48h, AKT gene transfection can increase the value of ARPE-19 cells (P<0.05),but after cultured for 72h, there was little difference (P>0.05). While the 10,20μg/ml of Conbercept group and 5% and 10% contained serum group,were lower than model cells group(P<0.05). After cultured for 72h,there was little or no difference in proliferation between model cell and control group(P>0.05).Results of the expression of VEGF in the supernatant of cells detected by ELISA shown that the model group was higher than that of normal cells group (P<0.05), serum and Conbercept group were significantly lower than the model group (P<0.05), no statistically significant difference between the blank serum group and model group cell difference (P>0.05). Conclusion The AKT gene transfection can induce proliferation of ARPE-19 cells and promote the expression of VEGF. The modified Zhu Jing Fang medicated serum has inhibitory effect on the proliferation and VEGF expression of ARPE-19 cells transfected with AKT gene. The effect of intervention on the generation of CNV may be played by suppressing the overexpression of VEGF. |
| keywords:Jiajian Zhujing prescription Containing serum ARPE-19 Gene transfection VEGF |
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