Study On The Chemical Constituents Of Patrinia Punctiflora And Its Effect On Improving Insulin Resistance In HepG2 Cells

Patrinia punctiflora belongs to the Patrinia genus in the Valerianaceae,mainly distributed in southern Henan,southwestern Shaanxi,Jiangsu and other places,and is a traditional dual-use resource for medicine and food.In the region of Zhejiang Changhua,P.punctiflora as folk medicine is used for antiphlogistic,anticoagulant,purulent and diuretic and in Tiantai Mountain,its whole herbs is used to treat appendicitis,in Guangxi,its root dip wine is used to treat falls,and leaf external application to wash sore poison.in the region of Jiangxi,its tender leaves are used as vegetables.According to literature research,Patrinia plants are often used in the formula for the treatment of diabetes.At the same time,the research group has previously discovered a series of chemical components with hypoglycemic activity from P.scabiosaefolia.,and screened for Hep G2 cell insulin resistance activity for P.punctiflora.It was found that at the same concentration,the effect of P.punctiflora extraction on promoting glucose absorption in Hep G2cells is more significant than P.scabiosaefolia.In order to deeply explore the hypoglycemic active ingredients of Patrinia plants,this study selected P.punctiflora from this genus for chemical composition and activity screening,as well as mechanism exploration,in order to provide effective basis for the deep development and utilization of Patrinia plants.The ethanol extraction method was used to obtain the whole herb extract of P.punctiflor a.D-101 Macroporous Resin,Silica gel,ODS and Sephadex LH-20 were selected as separation materials.In combination with Flash chromatography and Semipre-HPLC separation technologies,the chemical constituents were systematically studied by means of NMR,MS,IR,UV,ORD an d other spectral analysis methods（HSQC,HMBC,¹H-¹H COSY and NOESY）.In addition,a tot al of 36 compounds were isolated and identified by comparison between standard substances and literature,mainly lignans,iridoids,aromatic aldehydes and other compounds,as follows:（1S,3R,5S,7S,8S,9S）-1-methoxy-7-hydroxy-8-methyl-3,8-epoxy-Δ^4,11-dihyronepetane（1）,（1S,3R,5S,7R,8S,9S）-1-methoxy-7-hydroxy-8-methyl-3,8-epoxy-Δ^4,11-dihyronepetane（2）,patriscabioin M（3）,patriscabioin P（4）,8,9-didehydro-7-hydroxydolichodial（5）,patriscabioin Q（6）,jatamanin N（7）,patrirscabio in R（8）jatamanin Y（9）,patrinoside H（10）,patrinoside-aglucone（11）,stenopterin A（12）,（+）-n ortrachelogenin（13）,matairesionol（14）,（+）-isolariciresinol（15）,（7S,8R）-dihydrodehydrodiconiferyl alcoho（16）,（+）-lariciresinol（17）,（+）-syringaresinol（18）,（+）-medioresinol（19）,8-hydroxy-7’-epipi noresinol（20）,pinoresinol（21）,pinoresinol-4-O-β-D-glucopyranoside（22）,illiciumlignan C（23）,matairesinol 4’-O-β-D-glucopyranoside（24）,kaempferol-3,7-di-O-α-L-rahmnoside（25）,sinapic alde hyde（26）,alismoxide（27）,coniferaldehyde（28）,syringaldehyde（29）,vanillin（30）,1H-indole-3-c arbaldehyde（31）,ethyl caffeate（32）,benzyl-β-D-glucopyranoside（33）,methyl-β-D-glucopyranosid e（34）,（3S,5R,6S,7E）3,5,6-trihydroxy-7-megastigmen-9-one（35）,（-）-loliolide（36）.Compounds 4,6～10 are new iridoids.Insulin resistance model was established on Hep G2 cells with glucosamine,rosiglitazone was used as positive control drug,and the improvement effect and molecular mechanism of the chemical components of Patrinia punctiflora on hepatic insulin resistance were studied from the perspective of promoting glucose absorption,and the compounds of Patrinia punctiflora with the activity of improving insulin resistance were explored.The results showed that compounds 1,3,9,13,18,26～29,31～32 could promote the glucose uptake of IR-Hep G2 cells to different degrees.By comparing the MTT and GOD POD experiments,the effects of new compound 9 on the expression of some key proteins related to insulin resistance in Hep G2 cells were detected by Western blotting technique from the perspective of the optimal dose and multiple perspectives of the new compound.The results showed that compound 9 increased the protein expression levels of PI-3K,p-AKT,GLUT4 and p-GSK3β,and inhibited the expression of PEPCK and G6Pase proteins,which are key gluconeogenetic enzymes.These results suggest that compound 9 can improve insulin resistance by activating the PI-3K/AKT signaling pathway,promoting phosphorylated Akt to activate the downstream signaling molecule glucose transporter 4（GLUT4）to the plasma membrane,promoting glycogen synthesis,inhibiting gluconogenesis,and thereby improving insulin resistance.