The Chemical Constituents From Patrinia Villosa And Effects On Improving Insulin Resistance In HepG2 Cells

Patrinia villosa(Thunb.)Juss.,also known as Pandaozeng,is a perennial herb of Patrinia genus in Valerianaceae,mainly distributed in Guizhou,Hunan,Hubei and Henan provinces of China.The medicinal parts of Baijiangcao are P.villosa and Patrinia scabiosaefolia Fisch.ex Trev with roots.Its taste is bitter,slightly cold,with the effect of clearing heat and detoxifying,eliminating carbuncle discharge pus,removing blood stasis and relieving pain.In addition to the medicinal value,young leaves of P.villosa can be eaten as wild vegetables,which belongs to the resources of both medicine and food.Literatures showed that the research on Baijiangcao mainly focused on P.scabiosaefolia,while the research on P.villosa was less.Through activity screening,it was found that P.villosa extract had the activity of improving insulin resistance,but its active components were not clear.Therefore,in this paper,the chemical constituents of P.villosa were systematically studied,and the insulin-resistant Hep G2 cells were used as an experimental model in vitro to evaluate the improvement of insulin resistance activity and its mechanism of action of compounds isolated from P.villosa.Macroporous resin,silica gel and gel as separation materials were used,the monomer compounds were separated and purified by pressure,atmospheric and decompression column chromatography,rapid preparation liquid chromatograph and semipreparative high performance liquid chromatography(SemipreHPLC).A total of 32 compounds were isolated and identified from 70% ethanol extract of P.villosa by means of ultraviolet(UV),infrared(IR),nuclear magnetic resonance(NMR)and mass spectrometry(MS),which were as follows: 8,9-didehydro-7-hydroxydolichodial(1),patrirscabioin P(2),patrirscabioin M(3),(1S,3R,5S,7S,8S,9S)-1-methoxy-7-hydroxy-8-methyl-3,8-epoxy-Δ4,11-dihyronepetane(4),patrirscabioin Q(5),patrinoside-aglucone(6),7-O-trans-p-coumaroyl patrinoside(7),7-O-cis-p-coumaroylpatrinoside(8),2-O-trans-p-Coumaroyl loganin(9),2-O-cis-p-Coumaroyl loganin(10),patrinoside I(11),patrinoside(12),viburtinoside II(13),viburtinoside Ⅲ(14),nardostiridoid B(15),patrinoside J(16),patrinoside K(17),7-α-O-methylmorroniside(18),7-β-O-methylmorroniside(19),forsythide dimethyl ester(20),dihydrodehydrodiconiferyl alcohol(21),pinoresinol-4-O-glucoside(22),wasabiside A(23),dihydrodehydrodiconiferyl alcohol-4-O-β-D-glucopyranoside(24),(7S,8R)-1-(4-O-β-D-glucopyranosyl-3-methoxyphenyl-2-[4-(3-hydroxypropyl)-2,6-dimethox--yphenoxy]-1,3-propanediol(25),pinoresinol(26),matairesinol(27),1-(4′-hydroxy-3′-methoxyphenyl)-2-[4′′-(3-hydroxypropyl)-2′′,6′′-dimethoxyphenoxy]propane-1,3-diol(28),kaempferide 3-O-neohesperidoside(29),Kaempferol 3-O-[α-Lrhamnopyranosyl-(1→6)]-[α-L-rhamnopyrano-syl-(1→2)]-β-D glucopyranoside(30),betulinic(31),betulinic acid(32).Among them,compounds 2,5,8,11,16,17 were new compounds and compounds 7,9,10,13-15,29-32 were isolated from Patrinia genus for the first time.The results of activity screening showed that,compounds 22(pinoresinol-4-O-glucoside)and 23(wasabiside A)could promote glucose metabolism in insulin-resistant Hep G2 cells.Further study of the mechanism of action showed that compounds 22(PG)and 23(WA)promote the expression of PI-3K,p-AKT,GLUT4 and p-GSK3β proteins,and inhibit the expression of PEPCK and G6 Pase proteins,thus activating PI-3K/AKT signaling pathway,promoting glucose uptake and glycogen synthesis,indirectly inhibiting gluconeogenesis and improving insulin resistance.