熊果酸减轻LPS诱导的THP-1细胞损伤

Ursolic acid alleviated the LPS-induced damage in THP-1 cells

目的探讨熊果酸减轻脂多糖诱导的THP-1细胞损伤的作用及其机制。方法以脂多糖诱导的THP-1炎性细胞为模型,MTT法检测不同浓度的熊果酸(0.1、1、5、10、20、40和80μmol/L)对细胞增殖的影响,RT-PCR法检测TLR4、MCP-1和IL-6 mRNA的表达,ELISA法检测MCP-1和IL-6表达,Western blot法检测P65、磷酸化P65蛋白表达,荧光素酶报告系统检测核转录因子κB(NF-κB)活性。结果与对照组比较,LPS作用组能显著增高MCP-1、TLR4、IL-6 mRNA和MCP-1、IL-6、P65、磷酸化P65蛋白的表达并上调NF-κB活性(P<0.05);与LPS单独作用组比较,熊果酸(1和5μmol/L)干预组能够显著降低MCP-1、TLR4、IL-6mRNA和MCP-1、IL-6表达水平并下调NF-κB活性(P<0.05)。结论熊果酸可能是通过下调NF-κB活化减轻脂多糖诱导的THP-1细胞的损伤。

Objective To investigate the alleviating effect and mechanism of ursolic acid against LPS-induced dam-age in THP-1 cells. Methods THP-1 cells were exposed to 10 μg / L LPS for 20 h,ursolic acid of different con-centrations were added. Cell proliferation was tested by MTT,the expressions of TLR4,MCP-1 and IL-6 mRNA were detected by RT-PCR,enzyme-linked immunosorbent assay（ ELISA） was applied to detect the production of monocyte chemoattractant protein1（MCP-1）and interleukin-6（IL-6）,P65 and Phosphorylation-P65 were detec-ted on protein level using Western blot,the nuclear transcription factor kappa B（NF-kappa B）activity was detec-ted by luciferase report system. Results LPS group significantly increased MCP-1,TLR4,IL-6 mRNA and MCP-1,IL-6,P65,Phosphorylation-P65 proteins expression and enhanced NF-κB activity. Ursolic acid（1, 5 μmol / L） intervention groups significantly reduced MCP-1, TLR4, IL-6 mRNA expression and MCP-1, IL-6 pro-teins expression and inhibited NF-κB activity. Conclusions Ursolic acid may alleviate LPS-induced damage of THP-1 cells by reducing the NF-κB activity.