基于LPS炎症模型探究贝母素甲与贝母素乙对巨噬细胞Raw264.7的抗炎作用  

Exploring the Anti-inflammatory Effects of Peimine and Peiminine on Macrophage Raw264.7 Based on LPS Inflammation Model

作　　者：金泽兰 董云哲 李青 董莉莉 郭焕茹 严灿龙 王忠华[1] JIN Ze-Lan;DONG Yun-Zhe;LI Qing;DONG Li-Li;GUO Hua-Ru;YAN Can-Long;WANG Zhong-Hua(College of Biological&Environmental Sciences,Zhejiang Wanli University,Ningbo 315100,China;School of Life Sciences,Ningxia University,Yinchuan 750000,China;Ningbo Agricultural Technology Extension Station,Ningbo 315100,China)

机构地区：[1]浙江万里学院,生物与环境学院,宁波315100 [2]宁夏大学生命科学学院,银川750000 [3]宁波市农业技术推广总站,宁波315100

出　　处：《农业生物技术学报》2024年第10期2361-2370,共10页Journal of Agricultural Biotechnology

基　　金：浙江省中药材新品种选育重大科技专项(2021C02074);宁波市重点研发计划暨“揭榜挂帅”项目(2023Z120)。

摘　　要：贝母素甲与贝母素乙为百合科草本植物浙贝母(Fritillaria thunbergii)中的代表性生物碱类药用成分,对浙贝母抗炎功效发挥重要作用。为从细胞水平上进一步探究这两类生物碱的抗炎功效,本研究以脂多糖(lipopolysaccharide,LPS)诱导的小鼠(Mus musculus)单核巨噬细胞(Raw264.7细胞)为炎症模型,采用Griess法、ELISA、Western blot、qRT-PCR等方法分析贝母素甲和贝母素乙对LPS诱导的Raw264.7细胞分泌促炎因子一氧化氮(nitric oxide,NO)、肿瘤坏死因子-α(tumour necrosis factor-α,TNF-α)、白介素-6(interleukin-6,IL-6)、IL-1β、一氧化氮合酶(induciblenitric oxide synthase,iNOS)、环氧合酶-2(cyclooxygenase-2,COX-2)的影响。结果显示,贝母素甲与贝母素乙均能抑制Raw264.7细胞分化,减少LPS刺激产生的伪足,贝母素甲与贝母素乙中、高剂量组Raw264.7细胞上清液中NO、TNF-α、IL-6、IL-1β浓度显著降低(P<0.05,P<0.01)。贝母素甲与贝母素乙对Raw264.7细胞iNOS、COX-2蛋白表达均有抑制作用,且对于iNOS蛋白,贝母素乙的抑制作用高于贝母素甲,对于COX-2蛋白,贝母素甲的抑制作用高于贝母素乙。贝母素甲和贝母素乙均能显著降低Raw264.7细胞IL-6和IL-1β表达量。综上,贝母素甲和贝母素乙均有良好的抗炎效果。本研究为生物碱类物质对炎症的治疗提供一定的数据支持。Peimine and Peiminine are representative alkaloid medicinal component in the lily family herb Fritillaria thunbergii.They play an important role in the anti-inflammatory effect of F.thunbergii.In order to further investigate the anti-inflammatory effects of these 2 types of alkaloids at the cellular level,this study used lipopolysaccharide(LPS)induced Raw264.7 cells as an inflammatory model,Griess,ELISA,Western blot and qRT-PCR were used to analyze the effects of peimine and peiminine on LPS-induced secretion of pro-inflammatory factors nitric oxide(NO),tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),IL-1β,nitric oxide synthase(iNOS)and cyclooxygenase-2(COX-2)in Raw264.7 cells,the results showed that both peimine and peiminine could inhibit the differentiation of Raw264.7 cells and reduce the pseudopodia produced by LPS stimulation,NO,TNF-α,IL-6 and IL-1βin Raw264.7 cell supernatant of medium and high dose groups of peimine and peiminine significantly decreased(P<0.05,P<0.01).Peimine and peiminine inhibited the expression of iNOS and COX-2 protein in Raw264.7 cells,and the inhibition of peiminine on iNOS protein was higher than that of peiminine on COX-2 protein,the inhibitory effect of peiminine a was higher than that of peiminine.The expression of IL-6 and IL-1βin Raw264.7 cells was significantly decreased by both peimine and peiminine.In conclusion,both peimine and peiminine have good anti-inflammatory effect,which provides some data support for the treatment of inflammation by alkaloids.

关 键 词：贝母素甲 贝母素乙 RAW264.7 抗炎活性 

分 类 号：S326[农业科学—作物遗传育种]