丁苯酞对于星型胶质细胞炎症反应的保护作用及缝隙连接蛋白Cx43表达的影响

Protective Effect of Dl-3-n-butylphthalidle on Inflammation of Astrocytes and the Expression of Connexin 43

关键词:  丁苯酞  星形胶质细胞  脂多糖  炎症反应  缝隙连接蛋白Cx43

Key words:  butylphthalide  astrocytes  lipopolysaccharide  inflammatory response  connexin 43

摘要:

目的 研究丁苯酞（dl-3-n-butylphthalidle，NBP）对于脂多糖（lipopolysaccharide，LPS）诱导的星型胶质细胞（astrocytes，Ast）炎症反应的抑制作用和机制，同时观察其对于缝隙连接蛋白Cx43表达的影响。方法 Ast细胞采用梯度浓度的NBP处理，1 μg·mL^-1的LPS诱导炎症反应。试剂盒检测乳酸脱氢酶（lactate dehydrogenase，LDH）漏出量，CCK-8法检测细胞存活率，Griess法检测NO的释放水平，ELISA法检测培养基中炎症因子IL-1β、IL-6及TNF-α的表达水平。Western blotting法检测p65，p-IκB，Cx43的表达。免疫荧光染色法检测Cx43的分布。siRNA沉默Ast中Cx43的表达，LPS诱导炎症发生后，检测Ast的炎症因子释放以及细胞存活水平。结果 NBP可以降低Ast细胞中LDH的漏出，提高细胞存活率，抑制炎症因子的释放，同时抑制细胞中p65、p-IκB的表达。NBP可以同时抑制Ast中Cx43的表达。siRNA沉默Cx43后，Ast的炎症反应得到抑制。结论 丁苯酞可以抑制LPS诱导的Ast炎症反应，而Cx43与炎症反应的发生有关。丁苯酞抑制Ast炎症反应与抑制Cx43的表达有关。

Abstract:

OBJECTIVE To study the inhibition effect and mechanism of dl-3-n-butylphthalidle(NBP) on the inflammatory response of astrocytes(Ast) induced by lipopolysaccharide(LPS) and its effect on the expression of connexin 43(Cx43). METHODS Ast was treated with gradient concentration of NBP, and 1 μg·mL^-1 LPS induced inflammatory response. Lactate dehydrogenase(LDH) leakage was detected by kit, cell survival was detected by CCK-8, NO release was detected by Gries, IL-1β, IL-6 and TNF-α expression were detected by ELISA. Western blotting was used to detect the expression of p65, p-IκB and Cx43. The distribution of Cx43 was detected by immunofluorescence staining. SiRNA silenced the expression of Cx43 in Ast. After LPS induced inflammation, the release of inflammatory factors and cell survival were measured. RESULTS NBP could reduce LDH leakage, improve cell survival rate, inhibit the release of inflammatory factors, and inhibit the expression of p65 and p-IκB in Ast cells. NBP also inhibited the expression of Cx43 in Ast. After silencing Cx43 with siRNA, the inflammatory response of Ast was inhibited. CONCLUSION NBP can inhibit LPS induced Ast inflammatory response, and Cx43 is related to the occurrence of inflammatory response. The effect of NBP on Ast is related to the inhibition of Cx43 expression.