研究旨在探讨比较儿茶素和表儿茶素在小鼠肺泡巨噬细胞(MH-S)中的抗炎作用及其机制.利用脂多糖(LPS)刺激MH-S细胞建立炎症模型,用MTS法和LDH法检测儿茶素和表儿茶素对MH-S细胞活力的影响;ELISA法检测细胞上清液IL-1β、IL-6和TNF-α含量;qRT-PCR 法检测细胞Nos2、Il-6、Tnf-α、Grp78、Ire1α、Perk和Atf6 mR-NA 表达;Western Blot 法检测iNOS 蛋白表达.结果显示,6.25～100 μg/mL儿茶素和表儿茶素无显著细胞毒性,可浓度依赖地下调LPS诱导的IL-1β分泌和iNOS表达,还降低iNOS、IL-6以及内质网应激(ERS)相关蛋白IRE1α的mRNA转录水平.低浓度儿茶素的抗炎活性强于表儿茶素,高浓度则相反.研究表明,儿茶素和表儿茶素可以通过减少IL-1β、IL-6分泌,抑制iNOS蛋白表达发挥抗炎作用,其抗炎作用可能与抑制ERS有关.
Study on the anti-inflammatory effect and mechanism of catechin on mouse alveolar macrophages
The purpose of this study was to explore and compare the anti-inflammatory effects and the possible mechanisms of cate-chin and epicatechin in mouse alveolar macrophages(MH-S).Lipopolysaccharide(LPS)was used to stimulate MH-S cells to establish a cellular inflammation model.The effects of catechin and epicatechin on MH-S cell viability were detected by MTS and LDH assays.The contents of interleukin-1β(IL-1β),IL-6,and TNF-α in the supernatant were detected by enzyme-linked immunosorbent assay(ELISA).Real-time quantitative polymerase chain reaction(qRT-PCR)was used to detect the mRNA transcription levels of Nos2,Il-6,Tnf-α,Grp78,Ire1α,Perk,and Atf6.The protein expression of iNOS was detected by Western Blot.The results showed that 6.25-100 μg/mL of catechin and epicatechin had no significant toxicity on MH-S cells.Both catechin and epicatechin down-regulated the secretion of IL-1β and the protein expression of iNOS in LPS-induced MH-S cells in a concentration-dependent manner.Mean-while,catechin and epicatechin reduced the mRNA transcription levels of iNOS,IL-6 and endoplasmic reticulum stress(ERS)-related protein IRE1α.The anti-inflammatory activity of catechin was stronger than that of epicatechin at low concentrations,but catechin was lower than epicatechin at high concentrations.These results indicated that catechin and epicatechin had anti-inflammatory effects by re-ducing the secretion of IL-1β,IL-6 and the expression of iNOS,which might be related to the inhibition of ERS.
万方数据
维普
