熊果酸对脂多糖作用下肺上皮细胞炎症因子表达的影响及机制研究

Effect of ursolic acid on expression of inflammatory factors of pulmonary epithelial cells under lipopolysaccharide and its mechanism

摘要:

目的： 熊果酸（ursolic acid，UA）对脂多糖（lipopolysaccharide，LPS）作用下人肺上皮细胞来源的A549细胞的生物学活性影响及潜在机制。方法： CCK-8检测不同浓度（10、50、100、150、200 μg/mL）UA对A549细胞增殖的影响。此外，检测浓度为10 μg/mL的UA作用于A549细胞不同时间点对细胞的增殖影响。荧光定量PCR检测UA对LPS作用下A549细胞的白细胞介素（interleukin，IL）-1β、IL-6、肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）的mRNA表达。ELISA检测ERK、p38、JNK抑制剂对UA对LPS作用下A549细胞的IL-1β表达的影响，结果采用单因素方差分析。结果： UA能够显著抑制A549细胞的活性且呈浓度依赖性。CCK-8检测结果显示，UA作用A549细胞24 h后，10 μg/mL的UA能够明显抑制A549细胞的活性（P<0.05）。此外，UA能够明显抑制LPS诱导的TNF-α、IL-1β、IL-6的mRNA表达（P<0.05）。其中，对LPS诱导的IL-1β抑制作用最为明显。ELISA结果显示，ERK、JNK信号通路抑制剂作用下并不能影响UA抑制炎症的效果，但p38信号通路抑制剂能显著逆转UA对LPS诱导的IL-1β的抑制作用（P<0.05）。p38信号通路的激活剂却可以逆转这种抑制作用（P<0.05）。结论： UA能够抑制LPS诱导的IL-1β，且通过p38信号通路作用。

Abstract:

Objective: To observe the effect of ursolic acid (UA) on the biological activity of A549 cells derived from human pulmonary epithelial cells under lipopolysaccharide (LPS) and its potential mechanism. Methods: The CCK-8 method was used to detect the effect of UA (10, 50, 100, 150 and 200 μg/mL) at different concentrations on the proliferation of A549 cells. In addition, the effect of 10 ug/mL UA on the proliferation of A549 cells at different time points was also detected. The qPCR was used to detect the mRNA expression of interleukin (IL) -1β, IL-6 and tumor necrosis factor-α (TNF-α) in A549 cells under LPS. The effect of UA on the expression of IL-1β in A549 cells under LPS was detected by ELISA. Results: UA was able to significantly inhibit the activity of A549 cells. Results of CCK-8 showed that 10 μg/mL UA was able to significantly inhibit the activity of A549 cells after 24 hours of UA treatment (P<0.05). In addition, UA was able to significantly inhibit the mRNA expression of TNF-α, IL-1β and IL-6 induced by LPS (P<0.05). Among them, the inhibition effect of LPS-induced IL-1β was the most obvious. Results of ELISA showed that UA to inhibit inflammation was not influenced by ERK and JNK signaling pathway inhibitors, but p38 signaling pathway inhibitors was able to significantly reverse the inhibitory effect of UA on LPS-induced IL-1β (P<0.05). However, p38 signal pathway activator was able to reverse the inhibition effect (P<0.05). Conclusion: UA can inhibit the LPS-induced IL-1β and can exerts function through p38 pathway.