柿叶醋酸乙酯提取物对脂多糖刺激的小胶质细胞的抗炎及抗凋亡作用  

Anti-inflammatory and anti-apoptosis effect of extract of persimmon leaves on LPS-stimulated BV-2 cells

作　　者：林娟[1] 郭海彪[1] 苏诗韵 李楚源[1] 王德勤[1] LIN Juan;GUO Hai-biao;SU Shi-yun;LI Chu-yuan;WANG De-qin(Guangzhou Baiyun Mountain and Hutchison Whampoa Ltd., Guangzhou 510515)

机构地区：[1]广州白云山和记黄埔中药有限公司,广州510515

出　　处：《中南药学》2018年第11期1526-1530,共5页Central South Pharmacy

基　　金：2017年中医药国际合作专项(No.GZYYGJ2017037)

摘　　要：目的研究柿叶醋酸乙酯提取物(EAEPL)对脂多糖(LPS)刺激的小胶质细胞(BV-2)的炎症因子及凋亡细胞的表达作用,探讨EAEPL对脑卒中的抗炎及抗细胞凋亡的作用。方法采用LPS诱导的BV-2细胞作为体外神经炎症模型,用EAEPL进行干预;采用ELISA法检测白介素6(IL-6),白介素1β(IL-1β)和肿瘤坏死因子α(TNF-α)的含量变化;采用Western blot法检测IL-6、IL-1β、TNF-α、磷酸化蛋白激酶(p-AKT)、磷酸化-细胞外信号调节激酶(p-ERK)、凋亡基因Bax、B淋巴细胞瘤-2基因(Bcl-2)的蛋白表达水平。结果 LPS刺激BV-2细胞24 h之后,IL-6、IL-1β和TNF-α炎症因子细胞表达及蛋白表达水平明显升高(P <0.01,P <0.001),p-AKT、p-ERK蛋白表达水平明显降低(P <0.01,P <0.001),促凋亡蛋白Bax表达水平明显升高(P <0.001),抗凋亡蛋白Bcl-2表达水平明显降低(P<0.001);EAEPL预处理后,相对于模型组,炎症因子细胞表达水平降低,IL-1β在10、20μg·mL^(-1)时差异具有统计学意义(P <0.05),IL-6、TNF-α在20μg·mL^(-1)时差异具有统计学意义(P <0.05);炎症因子蛋白表达水平均明显下降(P <0.05,P <0.01);p-AKT、p-ERK蛋白表达水平升高,p-AKT在各浓度差异均有统计学意义(P <0.05,P <0.01,P <0.001);p-ERK在10、20μg·mL^(-1)时差异具有统计学意义(P <0.01);Bax蛋白表达水平明显下降(P <0.05,P <0.01);Bcl-2蛋白表达水平明显升高(P <0.05,P <0.01,P <0.001)。结论 EAEPL可调节LPS刺激的BV-2的炎症因子表达和激活AKT/ERK通路,具有抗炎和抗细胞凋亡作用,进一步阐明了EAEPL抗脑卒中的作用机制。Objective To determine the anti-inflammatory and anti-apoptosis effects of ethyl acetate extract of persimmon leaves(EAEPL) on stroke and the expression of inflammatory factors and apoptosis cells of EAEPL on lipopolysaccharide(LPS)-stimulated microglia(BV-2) cells.Methods LPS-stimulated BV-2 cells were used as neuroinflammation model in vitro.The cells were pretreated with EAEPL.The content of TNF-α, IL-1 β,and IL-6 was detected by ELISA, and the protein expression of TNF-α, IL-1 β,IL-6, p-AKT, p-ERK, Bax and Bcl-2 was examined by Western blot.Results After 24 h, LPS stimulation the cellular and protein expression level of TNF-α, IL-1 β,and IL-6 of BV-2 cells was increased significantly(P<0.01, P<0.001), the protein expression level of p-AKT, p-ERK, and Bcl-2 was decreased significantly(P<0.01, P<0.001), and the protein expression level of Bax was increased(P<0.001).Pre-treatment with EAEPL, as compared with the control group, the expression level of cell inflammatory factors was decreased, the level of IL-1 β had statistical difference at 10 and 20 μg·mL^-1(P<0.05), and the level of TNF-α, and IL-6 had statistical difference at 20 μg·mL^-1(P<0.05).The protein expression level of inflammatory factors was significantly decreased(P<0.05, P<0.01).The protein expression level of p-AKT, and p-ERK was increased, with significant difference at each concentration of p-AKT(P<0.05, P<0.01, P<0.001).p-ERK showed significant difference at 10 and 20 μg·mL^-1(P<0.01).The protein expression level of Bax was decreased(P<0.05, P<0.01).The protein expression level of Bcl-2 was increased(P<0.05, P<0.01, P<0.001).Conclusion EAEPL can regulate the expression of inflammation factors and activate the pathway of AKT/ERK on LPS-stimulated BV-2 cells, suggesting the anti-inflammatory and anti-apoptosis effect of EAEPL, and the mechanism of EAEPL against stroke.

关 键 词：柿叶醋酸乙酯提取物 脑卒中 小胶质细胞 炎症因子 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]