Study On The Screening Of Protecting Liver Active Components And Their Mechanism Of Action In Odontites Vulgaris Moench

Objective To clarify the material basis of Odontites vulgaris Moench,and to make a preliminary study on its liver-protecting mechanism.Methods The ethanol extract of Odontites vulgaris Moench was prepared into different polar parts of water,n-butanol,ethyl acetate and petroleum ether.First of all,the activity screening of different polar parts of Odontites vulgaris Moench was carried out by MTT experiment.Second,the use of modern advanced separation and purification methods to prepare monomer compounds from the active site.At the same time,UPLC-Q-Exactive was used to analyze the chemical components in the active parts of Odontites vulgaris Moench,and the material basis research of Odontites vulgaris Moench was completed.The monomer components in the active part were verified by MTT experiment.Finally,detect the effects of active phenylethanoid glycosides and iridoids in Hep G2 cells on hydrogen peroxide-induced LDH release,SOD activity and GSH content in Hep G2 cells,and screen common differences through RNA-seq,using q RT-PCR and Western blot methods to detect their effects on the expression of ID2,HSPA6,HSP70,MYLIP and RGS16 m RNA and protein,and preliminarily explore the mechanism of Odontites vulgaris Moench.Results 1)Different concentrations of the n-butanol and ethyl acetate parts of Odontites vulgaris Moench had protective effects on Hep G2 cell injury induced by hydrogen peroxide;2)17 monomer compounds were derived from n-butanol and ethyl acetate of Odontites vulgaris Moench 3)A total of 35 compounds were identified by UPLC-MS analysis,which preliminarily clarified the material basis of Odontites vulgaris Moench;4)Acteoside,Isoacteoside,Arenarioside,and Melampyroside can increase the content of SOD and GSH and reduce the release of LDH;5)Acteoside can significantly increase the expression of ID2,HSP70,MYLIP,RGS16 m RNA caused by hydrogen peroxide,and reduce the expression of HSPA6 m RNA;Isoacteoside can significantly increase the expression of HSP70,MYLIP,RGS16 m RNA caused by hydrogen peroxide,and reduced the expression of HSPA6 m RNA;Arenarioside and Melampyroside can significantly reduce the increase in the expression of ID2,HSPA6,HSP70,MYLIP and RGS16 m RNA caused by hydrogen peroxide.At the same time,the protein test results showed that Acteoside can increase the expression of ID2,RGS16 protein,and reduce the expression of HSPA6,HSP70,MYLIP proteins;Isoacteoside can reduce the expression of HSP70,MYLIP and RGS16 proteins;Arenarioside can reduce ID2,HSPA6,MYLIP,HSP70,RGS16 protein expression;Melampyroside can increase ID2 and HSPA6 protein expression,and reduce HSP70,MYLIP,RGS16 protein expression.Conclusion The active parts of Odontites vulgaris Moench for protecting the liver are mainly n-butanol and ethyl acetate.Its chemical components are mainly divided into phenethyl alcohol glycosides,flavonoids,iridoids and phenolic acids.Among them,phenethyl alcohol glycosides and iridoids may be the main active ingredients that play a hepatoprotective effect.