Analysis Of Chemical Constituents And Activity Study Of Brassica Chinensis

In this experiment,Crateva unilocalaris Buch.(C.unilocalaris)shoots was as experiment materials,the three different extracts were extracted with 80%methanol,80%ethanol and 80%acetone,respectively.The chemical constituents were identified by UHPLC-ESI-MS/MS,and antioxidant activities was determined.The main antioxidant was characterized by DPPH·-UHPLC-HRMS and ABTS·+–UHPLC–HRMS analysis.In addition,the effects of the three different extracts on inhibiting intracellular ROS in H₂O₂-induced Hep G2 cell were determined.The effects of ethanol extract from C.unilocalaris on brain damage and cognitive impairment induced by D-galactose in mice and its prevention of acute liver injury induced by acetaminophen were assessed in this study,the relative mechanism was analyzed by Western blot(WB).The main findings are as follows:(1)The acetone extracts contained the highest total phenolics,total flavonoids,saponins contents in the tree extracts,reaching to 39.51±2.55,29.93±1.20,144.56±4.90 mg/g DW.There are few reports on the analysis of the composition and the activity of C.unilocalaris.This study firstly investigates the main ingredients in C.unilocalaris.The main compounds were analyzed by UHPLC-ESI-HEMS/MS,and the results showed that 14 compounds,including 6 phenols and 7 saponins,were identified,chlorogenic acid is the main phenolic compound,and chikusetsusaponin Iva is main saponins.The quantitative results showed that 80%acetone extract had higher content of each component,with chlorogenic acid reaching 13.99 mg/g and chikusetsusaponin Iva reaching 29.16 mg/g.(2)The three different extracts of C.unilocalaris showed better antioxidant activity,and 80%acetone extract had better scavenging effect on DPPH·and ABTS·+,The IC₅₀ values were 23.59±0.13,22.90±0.68?g/m L,respectively.The main antioxidant activities in 80%acetone extract of C.unilocalaris were identified by DPPH·-UHPLC-HRMS and ABTS·+–UHPLC–HRMS analysis.After the reaction with DPPH·,the concentrations of six phenolic compounds decreased significantly with 8.76%-50.61%,while the decrease of saponins was not observed,after the reaction with ABTS·+,the concentrations of six phenolic compounds decreased significantly with 18.95%?50.88%,while the decrease of saponins was not observed,which indicted that phenolic compounds but not saponins contributed significantly to the DPPH or ABTS radical scavenging activity of C.unilocalaris shoots according to the results of DPPH·–UHPLC–HRMS and ABTS·+–UHPLC–HRMS analyses.It can be concluded from the results of cell experiment that three different extracts at 120?g/m L have good inhibitory effect on H₂O₂-induced ROS production in Hep G2 cells.(3)For food and health reasons,80%ethanol extract was chosen to be used in subsequent animal tests.D-galactose(D-gal)induced aging similar to natural senescence in mice,the prevention of the ethanol extract(CSE)of C.unilocalaris on D-galactose induced oxidative damage in brain and liver and cognitive impairment of mice was evaluated in this study.The results shown that CSE significantly increased the SOD,CAT,GSH-Px activities and decreased the MDA content.Based on the result of H&E staining and immunohistochemical analysis,D-gal damage the morphology and number of Neu N cells in neurons in brain,especially,hippocampus,of mice,and D-gal damage the liver of mice.However,CSE improved the morphology and number of Neu N cells and brain injury.The result of western blot shown that CSE active PI3K/Akt/Nrf2 signal pathway,up-regulated the expression of p-PI3K and p-Akt,which further promoted the Nrf2 nuclear translocation(immunofluorescence),the secondary antioxidant enzymes(HO-1,NQO1)was also elevated to alleviate the damage by oxidative stress.Four behavioral experiments have shown that D-gal produces symptoms similar to anxiety and reduced learning and memory,while high doses of ethanol extract(600mg/kg)significantly improve cognitive impairment caused by D-gal.(4)The prevention of CSE on acetaminophen(APAP)-induced acute liver injury was evaluated in this study.The result shown that CSE obviously improve the four-representative index(ALT,AST,ALP,LDH)in the plasma,in addition,CSE increase the antioxidant enzyme activities(SOD,GSH,CAT)and decrease MDA content.The histomathological results showed that contains central lobular necrosis,nucleolysis,pyknosis and nuclear rupture occurred was observed in the APAP-induced mice(model group),CSE treatment reduced the severity and severity of liver injury in a dose-dependent manner,especially in the high-dose group(600mg/kg).WB results showed that CSE activated the expression of related proteins Nrf2,NQO1,Bax,Bcl-2,Caspase-3.It was speculated that C.unilocalaris might alleviate the APAP-induced liver injury by alleviating oxidative stress and apoptosis...