溪黄草甲素对脂多糖诱导的炎性反应的作用及机制研究  

Effect and mechanism of rabdoserrin A on LPS-induced inflammatory response

作　　者：孙礼芹 李光霞 王瑞[1] 李医明[1] 钱菲 贾琦[1] SUN Li-qin;LI Guang-xia;WANG Rui;LI Yi-ming;QIAN Fei;JIA Qi(School of Traditional Chinese Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Institute of Interdisciplinary Integrative Medicine Research,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)

机构地区：[1]上海中医药大学中药学院,上海201203 [2]上海中医药大学交叉科学研究院,上海201203

出　　处：《中草药》2021年第15期4561-4568,共8页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金资助项目(81973458)。

摘　　要：目的研究溪黄草甲素对脂多糖(lipopolysaccharides,LPS)诱导的体内外炎性反应的作用及机制。方法 BALB/c小鼠ipLPS建立急性炎性反应模型,给予溪黄草甲素和地塞米松进行干预,检测各组小鼠血清中肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)和白细胞介素-6(interleukin-6,IL-6)的水平。体外建立LPS诱导的小鼠单核巨噬细胞RAW264.7炎性模型,采用XTT法考察溪黄草甲素对RAW264.7细胞存活率的影响;采用Griess法考察溪黄草甲素对RAW264.7细胞上清液中一氧化氮(nitric oxide,NO)水平的影响;采用ELISA法考察溪黄草甲素对RAW264.7细胞上清液中TNF-α和IL-6水平的影响;采用Western blotting法考察溪黄草甲素对RAW264.7细胞诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)、Janus激酶2(Janus kinase 2,JAK2)/信号转导和转录激活因子3(signal transducer and activator of transcription 3,STAT3)、核因子-κB(nuclearfactor-κB,NF-κB)、丝裂原活化蛋白激酶(mitogen-activatedproteinkinase,MAPK)信号通路相关蛋白表达的影响。结果溪黄草甲素显著抑制LPS诱导的急性炎性小鼠模型血清中TNF-α和IL-6水平(P<0.05、0.01、0.001)。溪黄草甲素(≤5μmol/L)对RAW264.7细胞存活率无明显影响;溪黄草甲素显著抑制LPS诱导的RAW264.7细胞NO分泌和上清液中TNF-α、IL-6水平,以及iNOS、磷酸化JAK2(p-JAK2)和磷酸化STAT3(p-STAT3)蛋白表达水平(P<0.05、0.01、0.001),但对NF-κB和MAPK信号通路相关蛋白表达无抑制作用。结论溪黄草甲素能够抑制LPS诱导的体内外炎性反应,其作用机制可能与抑制JAK2/STAT3信号通路有关。Objective To study the effect and mechanism of rabdoserrin A on lipopolysaccharides(LPS)-induced inflammatory response in vivo and in vitro. Methods BALB/c mice were ip LPS to establish an acute inflammatory response model, rabdoserrin A and dexamethasone were used for intervention, levels of tumor necrosis factor-α(TNF-α) and interleukin-6(IL-6) in serum of mice in each group were detected. LPS-induced inflammatory model of RAW264.7 cells was established in vitro, and XTT method was used to investigate the effect of rabdoserrin A on survival rate of RAW264.7 cells;Griess was used to investigate the effect of rabdoserrin A on nitric oxide(NO) level in supernatant of RAW264.7 cells;ELISA was used to investigate the effect of rabdoserrin A on levels of TNF-α and IL-6 in supernatant of RAW264.7 cells;Western blotting was used to investigate the effect of rabdoserrin A on expressions of inducible nitric oxide synthase(iNOS), Janus kinase 2(JAK2)/signal transducer and activator of transcription 3(STAT3), nuclear factor-κB(NF-κB) and mitogen-activated protein kinase(MAPK) signaling pathway related protein in RAW264.7 cells. Results Rabdoserrin A significantly inhibited the levels of TNF-α and IL-6 in the serum of acute inflammation mice model induced by LPS(P < 0.05, 0.01, 0.001). Rabdoserrin A(≤ 5 μmol/L) had no significant effect on survival rate of RAW264.7 cells;Rabdoserrin A significantly inhibited NO secretion, levels of TNF-α and IL-6 in supernatant, expressions of i NOS, phosphorylated JAK2(p-JAK2) and phosphorylated STAT3(p-STAT3) in RAW264.7 cells induced by LPS(P < 0.05, 0.01, 0.001), but had no inhibitory effect on expressions of NF-κB and MAPK signaling pathways related protein. Conclusion Rabdoserrin A can inhibit the inflammatory response induced by LPS in vivo and in vitro, and its mechanism may be related to the inhibition of JAK2/STAT3 signaling pathway.

关 键 词：溪黄草甲素 RAW264.7细胞 脂多糖 抗炎 JAK2/STAT3信号通路 

分 类 号：R285.5[医药卫生—中药学]