茺蔚子中木脂素类化学成分及肝细胞保护作用研究  

Lignans from Leonuri Fructus and their hepatoprotective effects

作　　者：刘娟汝 彭成[1,2] 何育霖 彭芳[1,2] 蒙春旺 熊亮 LIU Juan-ru;PENG Cheng;HE Yu-lin;PENG Fang;MENG Chun-wang;XIONG Liang(State Key Laboratory of Southwestern Chinese Medicine Resources,School of Pharmacy,Chengdu University of Traditional Chinese Medicine;Institute of Innovative Medicine Ingredients of Southwest Specialty Medicinal Materials,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China)

机构地区：[1]成都中医药大学药学院西南特色中药资源国家重点实验室 [2]成都中医药大学西南特色药材创新药物成分研究所,成都611137

出　　处：《天然产物研究与开发》2022年第10期1699-1706,1763,共9页Natural Product Research and Development

基　　金：国家自然科学基金(82022072,U19A2010);西南特色中药资源多维评价多学科交叉创新团队(ZYYCXTD-D-202209);特色中药资源多维评价与产品开发创新团队(2022C001)。

摘　　要：研究益母草Leonurus japonicus Houtt.果实茺蔚子中木脂素类化学成分及其肝细胞保护活性和作用机制。采用大孔吸附树脂、硅胶、HW-40F凝胶柱色谱及半制备液相色谱等对茺蔚子正丁醇提取部位进行分离纯化,根据理化性质及波谱数据鉴定化合物结构。本文从茺蔚子正丁醇提取部位分离鉴定了8个木脂素类化合物,分别为(+)-异落叶松脂醇-9′-O-β-D-葡萄糖苷(1)、(-)-异落叶松脂醇-9′-O-β-D-葡萄糖苷(2)、马先蒿木脂素(3)、(-)-异落叶松脂醇-4-O-β-D-葡萄糖苷(4)、异落叶松脂素(5)、phyllanglaucin B(6)、(-)-7 R,8 S-7′,8′-二羟基-二氢脱氢松柏醇-9-O-β-D-葡萄糖苷(7)、(+)-7 S,8 R-7′,8′-二羟基-二氢脱氢松柏醇-9-O-β-D-葡萄糖苷(8),化合物1~8均为首次从茺蔚子中分离得到。其次,采用MTT法考察化合物对对乙酰氨基酚(acetaminophen,APAP)诱导的HL-7702肝细胞损伤的保护作用,结果表明化合物1和3可显著提高APAP诱导肝细胞损伤的存活率;进而,采用Hoechst 33342荧光染色法和Western blot法观察活性最好的化合物3对细胞凋亡以及Bcl-2和Bax蛋白表达水平的影响。结果显示,与模型组相比,化合物3可使细胞核致密浓染现象变少,凋亡小体减少,同时明显促进Bcl-2蛋白表达,抑制Bax蛋白表达,并升高Bcl-2/Bax比值,表明化合物3的作用机制与调节Bcl-2和Bax蛋白表达量,抑制细胞凋亡有关。To study the chemical constituents from Leonuri Fructus(the fruits of Leonurus japonicus Houtt.)and their hepatocellular protective effects and mechanisms.The chemical compounds were isolated from the n-butanol extract of Leonuri Fructus by macroporous resin,silica gel,HW-40F column chromatography,and semi-preparative HPLC.Their structures were determined by physicochemical properties and spectroscopic data.Finally,eight lignans were isolated and identified as(+)-isolariciresinol-9′-O-β-D-glucopyranoside(1),(-)-isolariciresinol-9′-O-β-D-glucopyranoside(2),densispicoside(3),(-)-isolariciresinol-4-O-β-D-glucopyranoside(4),isolariciresinol(5),phyllanglaucin B(6),(-)-7 R,8 S-7′,8′-dihydroxy-dihydrodehydroconiferyl alcohol-9-O-β-D-glucopyranoside(7),and(+)-7 S,8 R-7′,8′-dihydroxy-dihydrodehydroconiferyl alcohol-9-O-β-D-glucopyranoside(8).Compounds 1-8 were isolated from Leonuri Fructus for the first time.Next,MTT assay was used to detect the protective effects of the isolates on acetaminophen(APAP)-induced hepatocyte injury.The results showed that compounds 1 and 3 significantly improved the survival rate of HL-7702 cells injured by APAP.Furthermore,Hoechst 33342 fluorescence staining and Western blot were carried out to observe cell apoptosis in HL-7702 cells and the expression level of Bcl-2 and Bax of compound 3 which has the best hepatoprotective effects.The results showed that compound 3 reduced the nuclear density of HL-7702 cells and the numbers of apoptotic bodies,significantly promoted the expression of Bcl-2 while inhibited the expression of Bax,and increased the ratio of Bcl-2/Bax expression compared with model group.It indicated that compound 3 played a role in suppressing apoptosis in HL-7702 cells by regulating the expression of Bcl-2 and Bax proteins.

关 键 词：茺蔚子 木脂素 肝细胞保护 作用机制 细胞凋亡 

分 类 号：R284.2[医药卫生—中药学]