【摘要】 目的 探讨补虚化浊方含药血清对氯化钴（CoCl₂）诱导的人视网膜色素上皮细胞（ARPE-19）自噬标志蛋白及血管内皮生长因子（VEGF）表达的影响。方法 将体外培养ARPE-19细胞分为正常组、缺氧组、康柏西普组、补虚化浊方组，正常组加入不含FBS的血清培养，缺氧组加入含CoCl₂ 100μmol/L的培养液+空白血清培养，康柏西普组加入含CoCl₂ 100μmol/L的培养液+20μg/mL康柏西普+空白血清培养，补虚化浊方组加入含CoCl₂ 100μmol/L的培养液+20%补虚化浊方含药血清培养。分别于培养24 h、48 h、72 h后，MTT法检测细胞增殖活性，ELISA法检测细胞上清液中VEGF含量；培养72 h后，Wester blot法检测细胞中自噬标志蛋白LC3BⅡ/Ⅰ、Beclin-1表达情况。结果 缺氧组细胞培养24 h、48 h、72 h的细胞活力吸光度值、上清液中VEGF含量和培养72 h后细胞中LC3BⅡ/Ⅰ、Beclin-1蛋白相对表达量均明显高于其他组（P均<0.05）,补虚化浊方组、康柏西普组、正常组间细胞活力吸光度值、上清液中VEGF含量和细胞中LC3BⅡ/Ⅰ、Beclin-1蛋白相对表达量比较差异均无统计学意义（P均>0.05）。结论 补虚化浊方可能是通过抑制自噬的激活，进而下调VEGF的表达来阻断脉络膜新生血管的发生。更多还原

【Abstract】 Objective It is to investigate the effects of the drug-containing serum of decoction for tonifying deficiency and resolving turbidity on the expression of autophagy marker protein and vascular endothelial growth factor（VEGF） in human retinal pigment epithelial cells（ARPE-19） induced by cobalt chloride（CoCl₂）. Methods The ARPE-19 cells cultured in vitro were divided into normal group, hypoxia group, Conbercept group and group of decoction for tonifying deficiency and resolving turbidity. The normal group was cultured with serum without FBS, the hypoxic group was cultured with culture medium containing CoCl₂ 100 μmol/L+blank serum, the Conbercept group was cultured with culture medium containing CoCl₂ 100 μmol/L+20 μg/mL Conbercept+blank serum, and the group of decoction for tonifying deficiency and resolving turbidity was cultured with culture medium containing CoCl₂ 100 μmol/L+20% drug-containing serum of decoction for tonifying deficiency and resolving turbidity. The cell proliferation activity was detected by MTT and the content of VEGF content in the cell supernatant was measured by ELISA after 24 h, 48 h and 72 h of culture, the expression of autophagy marker proteins LC3BⅡ/Ⅰ and Beclin-1 was measured by Wester blot after 72 h of culture. Results The absorbance values of cell viability, the content of VEGF in the supernatant after culture for 24 h, 48 h and 72 h, and the relative expression of LC3BⅡ/Ⅰ and Beclin-1 protein in the cells after cultured for 72 hours in the hypoxic group were significantly higher than those in the other groups（all P<0.05）, there was no statistically significant difference in the absorbance values of cell viability, the content of VEGF in the supernatant and the relative expression of LC3BⅡ/Ⅰ and Beclin-1 protein in the cells among normal group, Conbercept group and group of decoction for tonifying deficiency and resolving turbidity（all P>0.05）. Conclusion Decoction for tonifying deficiency and resolving turbidity may block the occurrence of choroidal neovascularization by inhibiting the activation of autophagy to down-regulate the expression of VEGF.更多还原