腺梗豨莶PPARγ激动活性成分及其抗炎作用研究  

Diterpenoids as PPARγ agonists from Siegesbeckia pubescens and their anti-inflammatory effects in vitro

作　　者：高丽娟[1] 徐世芳[1] 李晓誉[1] 黄文康 郝淑娟[1] 叶益萍 GAO Li-juan;XU Shi-fang;LI Xiao-yu;HUANG Wen-kang;HAO Shu-juan;YE Yi-ping(Key Laboratory of Neuropsychiatric Drug Research of Zhejiang Province,Institute of Materia Medica,Zhejiang Academy of Medical Sciences,Hangzhou 310013,China)

机构地区：[1]浙江省神经精神疾病药物研究重点实验室浙江省医学科学院药物研究所

出　　处：《中国中药杂志》2019年第23期5191-5197,共7页China Journal of Chinese Materia Medica

基　　金：浙江省中医药科技计划项目(2016ZB028)

摘　　要：该研究从腺梗豨莶Siegesbeckia pubescens水提物(SPA)中筛选、分离和鉴定PPARγ激动活性成分,并探讨它们的体外抗炎活性。通过瞬时转染PPARγ重组质粒的293T细胞模型,采用双荧光素酶报告基因法筛选、分离SPA中的PPARγ激动活性成分,并以色谱技术或光谱技术鉴定化学结构;进一步通过TNF-α诱导HT-29细胞的体外溃疡性结肠炎(UC)模型,以RT-PCR检测腺梗豨莶PPARγ激动活性成分对IL-1β,IL-8和TNF-αmRNA表达的影响,并以ELISA法检测它们对IL-8分泌的影响。结果显示,从腺梗豨莶具有PPARγ激动活性的D50组分中分离并鉴定了5个已知的二萜类化合物,分别鉴定为奇壬醇(1),豨莶精醇(2),对映-2-酮-15,16,19-三羟基海松烷-8(14)-烯-19-O-β-D-吡喃葡萄糖苷(3),豨莶苷(4)和对映-2β,15,16,19-四羟基海松烷-8(14)-烯-19-O-β-D-吡喃葡萄糖苷(5)。这5个化合物均具有显著的PPARγ激动活性且呈明显地浓度依赖性(P<0. 01)。此外,化合物1显著抑制IL-1βmRNA的表达和IL-8分泌(P<0. 001);化合物2和3均显著抑制IL-8,IL-1β,TNF-αmRNA的表达以及IL-8分泌(P<0. 01或P <0. 001);化合物4显著抑制IL-1β和TNF-αmRNA的表达(P <0. 01或P<0. 001);化合物5能显著抑制IL-1βmRNA表达(P <0. 001)。综上所述,腺梗豨莶中分离得到的二萜类化合物1~5具有PPARγ激动活性和潜在的体外抗UC作用。This study aims to investigate the PPARγ agonists isolated from the aqueous extract of Siegesbeckia pubescens( SPA) and their anti-inflammatory activities in vitro. The 293 T cells transfected transiently with PPARγ recombinant plasmid were used as a screening model to guide the isolation of PPARγ activitating components,and then PPARγ activities were measured by double luciferase reporter gene assay. The chemical structures were identified by chromatography or spectroscopic techniques. Furthermore,a UC inflammatory model in vitro was established on HT-29 cells by stimulating with TNF-α. The mRNA levels and secretion of proinflammatory cytokines on HT-29 cells,such as IL-1β,TNF-α,IL-8,were detected by RT-PCR and ELISA. The results showed that five diterpenoids were obtained from the fraction D50 with the strongest PPARγ activity among others in SPA,and determined as kirenol( 1),darutigenol( 2),enantiomeric-2-ketone-15,16,19-three hydroxypinomane-8( 14)-ene-19-O-β-D-glucoside( 3),darutoside( 4),enantiomeric-2-β,15,16,19-four hydroxypinomane-8( 14)-ene-19-O-β-D-glucoside( 5),respectively. All the compounds exhibited active effects on PPARγ in a concentration-dependent manner( P<0. 01). In addition,compound 1 significantly inhibited the expression of IL-1β mRNA and secretion of IL-8 on HT-29 cells inflammation model( P<0. 001);both compounds 2 and 3 effectively inhibited the expression of IL-1β,TNF-α,IL-8 mRNA and secretion of IL-8( P<0. 01 or P<0. 001),although at different extent;compound 4 significantly inhibited the expression of IL-1β and TNF-α mRNA( P<0. 01 or P<0. 001),while compound 5 inhibited the expression of IL-1β mRNA obviously( P<0. 001). In conclusion,the diterpenoids 1-5 isolated from S. pubescens have the PPARγ activation activities and potential effects of anti-UC in vitro.

关 键 词：腺梗豨莶 溃疡性结肠炎 PPARΓ TNF-Α IL-1Β IL-8 

分 类 号：R285[医药卫生—中药学] R284[医药卫生—中医学]