艳山姜挥发油调控NF-κB信号抑制LPS诱导的视网膜 Müller细胞炎症反应

Inhibitory Effect of Essential Oil from Aplinia zerubet Fruits on LPS-Induced Inflammation in Retinal Müller Cells by Regulating NF-κB Signal

目的:探索艳山姜挥发油对LPS诱导的视网膜Müller细胞炎症反应的保护作用及其作用机制。方法:采用LPS诱导视网膜Müller细胞发生炎症反应,给予低、高浓度艳山姜挥发油进行干预,分组为空白对照组、模型组、艳山姜挥发油低浓度组(1μg/L)、艳山姜挥发油高浓度组(5μg/L)。Western Blot法、ELISA法检测Müller细胞中炎症因子TNF-α、IL-6、IL-1β的表达和分泌。Western Blot法检测NF-κB信号分子TLR4、IκBα、p-IκBα、P65、p-P65蛋白表达,IF法观察P65入核情况。进一步采用NF-κB特异性抑制剂PDTC处理Müller细胞后,检测IκBα、p-IκBα、P65、p-P65、TNF-α、IL-6、IL-1β蛋白表达。结果:艳山姜挥发油能抑制LPS诱导Müller细胞炎症因子TNF-α、IL-6、IL-1β的合成与释放,抑制IκBα、p-IκBα、P65、p-P65蛋白表达,改善炎症反应。采用NF-κB特异性抑制剂PDTC干预后可抑制上述蛋白表达,且艳山姜挥发油+PDTC组与PDTC组比较无明显差异。结论:艳山姜挥发油可抑制LPS诱导的大鼠视网膜Müller细胞炎症反应,其作用机制与NF-κB信号的激活相关。

Objective:To investigate the protective effect and mechanism of essential oil from Aplinia zerubet fruits on LPS-induced retinal Müller cells inflammatory response.Methods:The inflammatory response of retinal Müller cells was induced by LPS,and low and high doses of essential oil from Aplinia zerubet fruits were administered for intervention.The rats were divided into the normal control group,model group,essential oil from Aplinia zerubet fruits low-dose group(1 μg/L) and high-dose group(5 μg/L).The expressions and secretions of inflammatory cytokines TNF-α,IL-6 and IL-1β in Müller cells were detected by Western Blot and ELISA.The protein expressions of NF-κB signaling molecules TLR4,IκBα,p-IκBα,P65 and p-P65 were detected by Western Blot,the nuclear situation of P65 was observed by IF method.The expressions of IκBα,p-IκBα,P65,p-P65,TNF-α,IL-6 and IL-1β were detected in Müller cells treated with PDTC,a specific inhibitor of NF-κB.Results:The essential oil of Aplinia zerubet fruits could inhibit the synthesis and release of inflammatory factors TNF-α,IL-6 and IL-1β in Müller cells induced by LPS,inhibit the proteins expressions of IκBα,p-IκBα,P65 and p-P65,improve the inflammatory response.NF-κB specific inhibitor PDTC could inhibit the expressions of the above proteins,and there was no significant difference between the volatile oil combined with PDTC group and PDTC group.Conclusion:The essential oil of Aplinia zerubet fruits can inhibit LPS-induced inflammation of Müller cells in the retina of rats,and the mechanism is related to the activation of NF-κB signal.