Chemical Constitutes From The Roots Of Juglans Manshurica Maxim. And Their Anti-tumor Effects On Lung-cancer Cells

Juglans mandshurica Maxim.is a deciduous tree of the genus Juglans in the family Juglandaceae,that is widely distributed in Northeast and North China.The roots,stem barks,leaves and fruits of this plant had been used as a tranditional medicine with effects of heat-clearing and detoxicating,and warming the kidney-yang.Previous phytochemical and pharmacological investigations revealed the presence of quinones,diarylheptanes,flavonoids and triterpenoids isolated from this plant and exhibited anti-tumor,anti-oxidant,anti-bacterial and analgesic activities.In order to elucidate the chemical constituents and pharmacological activity of this plant,the study was conducted in the Et OAc fraction of ethanol extract from the roots of J.mandshurica,using silica gel column and preparative middle pressure liquid chromatography.Twenty-five compounds were isolated and identified by NMR and other spectroscopic methods,including thirteen diarylheptanes: galeon(1),(6R)-3’’,4’-epoxy-6-hydroxy-7-(4’’-hydroxyphenyl)-1-(3’-methoxyphenyl)heptan-5-one(2),myricatomentogenin(3),juglanin A(4),(5R)-2’,5-dihydroxy-3’,4’’-dimethoxy-3’’,4’-oxo-1,7-diphenyl-5-heptanol(5),(5R)-2’,4’’,5-trihydroxy-3’-methoxy-3’’,4’-oxo-1,7-diphenyl-5-heptanol(6),juglanin B(7),(R)-4-(5-hydroxy-7-(4-hydroxyphenyl)-heptyl)-2-methoxy-phenol(8),3,7-dihydroxy-1-(4-hydroxyphenyl)-7-(4-hydroxy-3-methoxyphenyl)heptane(9),rhoiptelol C(10),3(R)-hydroxy-1,7-bis(4-hydroxy-3-methoxypheyl)heptane(11),1,7-bis(4-hydroxy-3-methoxyphenyl)-3-heptanone(12),rhoiptelol B(13),eight quinones: regiolone(14),botrytone(15),5,8-dihydroxy-4-methoxy-1-tetralone(16),5-methoxy-4-hydroxy-α-tetralone(17),2-methoxy juglone(18),3-methoxy juglone(19),5-hydroxy-2-hydroxymethyl-1,4-4-naphthoquinone(20),1,3-dimethoxycarbony-8-hydroxy-9,10-anthraquinone(21),three flavonoids:(-)-naringenin(22),dihydrokaempferol(23),kaempferol(24)and one triterpene:betulinic acid(25).Among them,compound 17 was the first time to be reported in the family Juglandaceae,20 was the first time to be identified from the genus Juglans,and 12 and 16 were obtained from J.mandshurica for the first time.The twenty-five isolated compounds were screened for in vitro anti-lung cancer activity by the MTT method.The experimental results showed that compounds 18,20 and 21 showed significant inhibitory effects on the proliferation of human lung cancer H1299,A549 cells.In addition,compound 18 inhibitited proliferatin of human lung cancer H1299, A549 cells,and promoted apoptosis by decreasing the expression level of anti-apoptotic protein Bcl-2,and increasing the expression levels of pro-apoptotic protein Bax,cleaved caspase-3 and cleaced-PARP.