Study On The Effects Of Incarvillea Arguta Royle On Liver Protection,Lithagogue,Choleretic And Its Chemical Constituents

Objection:This study used the classic liver injury,cholesterol gallstone and choleretic model to investigate the role of Incarvillea arguta Royle in liver protection,choleretic,and row of stones;UHPLC-Q-TOF-MS/MS Qualitative analysis of chemical constituents in Incarvillea arguta Royle.Methods:(1)The model of acute liver injury in mice was induced by intragastric ANIT(60 mg·kg-1)olive oil solution.The hepatoprotective effects of total extracts and total alkaloids were studied and serum glutamate-acetone was detected.Acid transaminase(ALT),aspartate aminotransferase(AST),alkaline phosphatase(ALP),total bile acid(TBA),total bilirubin(TBIL)and free bilirubin(DBIL);Superoxide dismutase(SOD),malondialdehyde(MDA)and glutathione peroxidase(GSH)in tissue homogenate,FXR,CYP7A1,BSEP,MRP2 mRNA in liver tissue by RT-PCR The expression level,hematoxylin-eosin(HE)staining to observe the pathological changes of liver tissue explored the hepatoprotective mechanism of the I.argute Royle;(2)Feeding mice with high-fat diet to produce a cholesterol stone model to study the stone-removing effect of stone grass,detecting TC,HDL-C,LDL-C,the level of triglyceride(TG),and measuring the relative expression of LXR-?,SRBI mRNA to study the stone-removing mechanism of the stone.(3)The normal choledochal drainage method was used to study the choleretic effect of calculus,and the biliary mechanism of calculus(TBIL)and TC in bile was studied to study the choleretic mechanism of calculus.(4)UHPLC-Q-TOF-MS/MS,YMC-ULtraHT Pro C18 column(100 mm × 3 mm,2 ?m)was eluted with a mobile phase gradient of acetonitrile-water solution;mass spectrometry was performed using an electrospray(ESI)ion source.Data were collected in negative ion mode,and the components were identified by retention time,accurate molecular ion peak and secondary mass spectrometry fragmentation.The flow rate is 0.4 mL·min-1,the column temperature is 30 ° C,and the detection wavelength is 254 nm.Result:(1)The ALT,AST,ALP,TBA,TBIL and DBIL levels in the serum of the liver can significantly reduce the levels of SOD and GSH in the liver homogenate of mice,and decrease the level of MDA,indicating the grass Anti-liver injury can be induced by inhibiting lipid peroxidation and scavenging free radicals;the mechanism of anti-liver injury by sarcophagus may be up-regulated by stimulating FXR expression,and enhanced expression of FXR inhibits the expression of bile acid synthase CYP7A1,acting on bile acids At the same time,the transcription of BSEP and MRP2 on the hepatocyte membrane is increased,which acts on the bile acid transport link.The two links work together to alleviate the cholestasis.(2)The high-dose group and the high-dose total alkaloid group significantly increased HDL-C levels and significantly decreased TC and TG levels,indicating that it may reduce cholesterol production rate by lowering cholesterol and lowering phospholipid levels;The mechanism of anti-liver injury of total alkaloids in grass may be to down-regulate the expression of LXR-?,and the decreased expression of LXR-? reduces the expression of SRBI,which makes the expression of SRBI insufficient and reduces the rate of stone formation.(3)Compared with normal rats,high-dose group of I.argute Royle and high-dose alkaloids could significantly increase the flow of bile in normal rats and reduce the levels of TBIL and TC in bile,but there was no significant change in TBA content.The relative expression of MRP2 and BSEP mRNA in the high dose group was significantly increased.The relative expression of LXR-? mRNA in the liver tissue of the alkaloid high dose group was significantly decreased,and the expression of BSEP mRNA was significantly increased.(4)UHPLC-Q-TOF-MS/MS technique was used to identify seven alkaloids in the I.argute Royle,namely O-methylcorypalline ? N-methylcorydaldine ? Boldine dimethyl ether?Lysicamine?Berbine ?Piperine,two flavonoids.Ingredients are Quercetin and Luteolin.Conclusion: The I.argute Royle has significant liver-protecting,treatment of cholesterol stonesand choleretic effects.Its liver-protecting mechanism is to increase the synthesis and transport of bile acids.The mechanism of treatment of cholesterol stones is to lower cholesterol.The content,reducing the level of phospholipids and increasing the content of HDLC to reduce the incidence of cholesterol stones.Its choleretic mechanism is to increase bile flow and change bile components.UHPLC-Q-TOF-MS/MS method can quickly identify the chemical components in the stone,simple and fast,and lay a certain foundation for the study of I.argute Royle.