前列欣胶囊对脂多糖诱导RAW264.7细胞炎症的抑制作用  

The anti-inflammatory of the butanol part of Qianliexin Capsule on LPS induced RAW264.7 cells

作　　者：陈龙 刘册家[2,3] 贾传青 穆岩 段文娟 臧凌鹤 于宗渊 王晓 CHEN Long;LIU Cejia;JIA Chuanqing;MU Yan;DUAN Wenjuan;ZANG Linghe;YU Zongyuan;WANG Xiao(Shandong Analysis and Test Center,Shandong Key Laboratory of TCM Quality Control Technology,Qilu University of Technology(Shandong Academy of Sciences),Jinan 250014,China;School of Pharmacy,Shandong University,Jinan 250012,China;Shandong Hongjitang Pharmaceutical Group Co.,Ltd.,Jinan 250103,China;School of Pharmacy,Shandong University of Traditional Chinese Medicine,Jinan 250355,China;School of Life Science and Biopharmaceutics,Shenyang Pharmaceutical University,Shenyang 110016,China)

机构地区：[1]齐鲁工业大学(山东省科学院),山东省分析测试中心,山东省中药质量控制技术重点实验室,山东济南250014 [2]山东大学药学院,山东济南250012 [3]山东宏济堂制药集团股份有限公司中药研究院,山东济南250103 [4]山东中医药大学药学院,山东济南250355 [5]沈阳药科大学生命科学与生物制药学院,辽宁沈阳110016

出　　处：《沈阳药科大学学报》2022年第10期1257-1262,共6页Journal of Shenyang Pharmaceutical University

基　　金：国家十三五重点研发计划(2017YFC1702704);山东省济南市泉城“5150”引才倍增计划(2017017);山东省重大科技创新工程项目(2021CXGC010508)。

摘　　要：目的 探讨前列欣胶囊正丁醇部位(the butanol part of Qianliexin Capsule, QLXB)对脂多糖(lipopolysaccharide, LPS)诱导小鼠巨噬细胞RAW264.7细胞分泌炎症细胞因子的影响,并探讨其对核因子-κB(nuclear factor kappa B,NF-κB)信号通路中相关蛋白的干预作用,初步明确QLXB的抗炎作用机制。方法 建立LPS诱导的RAW264.7细胞炎症模型,采用12.5、25、50 mg·L^(-1)剂量的正丁醇部位干预细胞24 h后,Griess法检测上清液中一氧化氮(nitric oxide, NO)含量;酶联免疫吸附法(enzyme-linked immunosorbent assay, ELISA)检测细胞上清液中肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白介素-1β(interleukin-1β,IL-1β)、白介素-6(interleukin-6,IL-6)和前列腺素E2(prostaglandin E2,PGE_(2))的含量;Western blot法检测Toll样受体4(Toll-like receptor 4,TLR4)、环氧化酶-2(cyclooxygenase-2,COX-2)、磷酸化的核转录因子p65 (phosphorylated nuclear factor kappa B p65,p-NF-κB p65)、磷酸化核因子-κB抑制蛋白激酶α/β(phosphorylated inhibitor of kappa B kinaseα/β,p-IKKα/β)和一氧化氮合酶(inducible nitric oxide synthase, iNOS)的表达情况。结果 与正常对照组比较,模型组RAW264.7细胞上清液中NO、IL-6、IL-1β、TNF-α和PGE_(2)的分泌水平显著升高,前列欣胶囊中正丁醇部位呈剂量依赖性降低细胞上清中上述炎症因子的分泌水平;在蛋白水平上,与模型组比较,TLR4、COX-2、p-NF-κB p65、p-IKKα/β和iNOS蛋白表达均显著升高,前列欣胶囊中正丁醇部位能有效抑制上述蛋白的表达,并呈剂量依赖性。结论 前列欣胶囊中正丁醇部位可能通过抑制NF-κB信号通路的激活而减轻LPS诱导的RAW264.7细胞炎症反应。Objective To preliminary explore the anti-inflammatory mechanism of the butanol part of Qianliexin Capsule(QLXB),to test the effects on the inflammatory cytokines of lipopolysaccharide(LPS) induced mice macrophage 264.7 cells, and to explore their intervention effect in the proteins associated with Nuclear factor kappa B(NF-κB).Methods The LPS-induced RAW264.7 cell inflammation model was established, and nitric oxide(NO) in cell supernatant was detected by Griess method after QLXB intervention with 12.5,25 and 50 mg·L^(-1)for 24 h;Tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),interleukin-6(IL-6) and prostaglandin E2(PGE_(2)) were detected by Enzyme-linked immunosorbent assay(ELISA);The expressions of Toll-like receptor 4(TLR4),cyclooxygenase-2(COX-2),phosphorylated nuclear factor kappa B p65(p-NF-kB p65),phosphorylated inhibitor of kappa B kinase α/β(p-IKKα/β) and inducible nitric oxide synthase(iNOS) were detected by Western blot.Results Compared with normal control group, the secretion levels of NO,IL-6,IL-1β,TNF-α and PGE_(2)in the model group increased significantly, QLXB had reduced the concentration significantly.For the protein expressions, TLR4,COX-2,p-NF-kB p65,p-IKKα/β and iNOS protein expressions in the model group were significantly higher compared with the control group.The QLXB effectively inhibited the expressions of the above proteins and showed dose-dependent property.Conclusion The QLXB may reduce the inflammatory response of RAW264.7 cells induced by LPS by inhibiting the activation of the NF-κB inflammatory signaling pathway.

关 键 词：RAW264.7细胞 炎症因子 NF-ΚB信号通路 

分 类 号：R96[医药卫生—药理学]