Study On Pharmacological Activities And Mechanisms Of Action Of Sesquiterpenoids Of Panax Ginseng

At present,the research on ginseng mainly focuses on the water-soluble components and activities such as saponins,proteins and polysaccharides,while the research on the fat-soluble parts of ginseng is less.The research group studied the Sesquiterpenoids in the fat-soluble parts of ginseng,established the extraction method of Sesquiterpenoids of Panax Ginseng?SPG?.The experimental research on the activity and mechanism of antidepressant-like and anti-acute liver injury was carried out.The main research contents and results were as follows:1.Establishment and composition analysis of extraction methodsIn this paper,the essential oil of ginseng was extracted by supercritical CO₂ extraction system,and the sesquiterpenoids compounds in the essential oil was refined by steam distillation to collection of distillates for 170-350 min,namely SPG.The sesquiterpenoids compounds was analyzed by gas chromatography-mass spectrometry?GC-MS?,and a total of 21 structures were identified.2.Study on the antidepressant mechanism of SPGThe mouse depression model was induced by lipopolysaccharide?LPS?to explore the antidepressant-like activity and mechanism of action of SPG.ICR mice were given SPG?0.25and 1 mg/kg?for 7 days by oral administration,and LPS?0.5 mg/kg?was intraperitoneally injected 1 hour after SPG administration on the last day.Behavioral experiments were performed 24 hours after LPS administration,and serum and hippocampus of mice were obtained after the behavioral experiment.The relevant indexes in serum and hippocampus were determined according to the kit instructions.At the same time,the antidepressant-like mechanism of SPG was analyzed by detecting the western-blot results of hippocampus.The results showed that with LPS model group,SPG effectively reduced the immobility time of mice in tail suspension test?TST?and forced swimming test?FST?;SPG significantly reduced the levels of tumor necrosis factor-??TNF-??and interleukin-6?IL-6?in the serum.In addition,Western-blot results showed that SPG up-regulated the expression of brain-derived neurotrophic factor?BDNF?,tropomyosin-related kinase B?TrkB?and sirtuin type 1?Sirt1?,and down-regulated nuclear factor-?B?NF-?B?and?B-?inhibitor?I?B-??.In summary,SPG?0.25and 1 mg/kg?had significant antidepressant-like effect,and its mechanism of action was related to inhibition of hippocampal inflammation and increase of oxidative stress and neurotrophic factors.3.Study on the mechanism of SPG to protect acute liver injuryThree different models of acute liver injury,including carbon tetrachloride?CCl₄?model,D-galactosamine?D-GalN?/LPS model and acetaminophen?APAP?model were used to explore the protective effect and mechanism of SPG on the acute liver injury.?1?Establishment of CCl₄ acute liver injury model.The CCl₄ acute liver injury model was used to explore the pharmacological activity and mechanism of SPG protection against acute liver injury by CCl₄-induced.ICR mice were given SPG?2.5 and 10 mg/kg?for 7 days by oral administration,and CCl₄?0.2%?was intraperitoneally injected 1 hour after SPG administration on the last day.After 24 hours of CCl₄ administration,the serum and liver of the mice were collected.The relevant indexes in serum and liver tissues were determined according to the kit instructions.The mechanism of SPG protecting CCl₄-induced acute liver injury was analyzed by H&E staining,Western-blot and RT-PCR.The results showed that SPG significantly decreased the levels of aspartate aminotransferase?AST?,alanine aminotransferase?ALT?,IL-6,interleukin-1??IL-1??and TNF-?in the serum;the content of glutathione?GSH?,superoxide dismutase?SOD?and catalase?CAT?was increased,and the content of malondialdehyde?MDA?was lowered in liver tissue compared with the CCl₄ model group.In addition,it was found from the H&E staining analysis that SPG could significantly improve the CCl₄-induced apoptosis and inflammatory infiltration of liver cells.Besides,western-blot results showed that SPG could inhibit the expression of c-Jun N-terminal kinase?p-JNK?,extracellular regulatory protein kinase?p-ERK?,p-p38,NF-?Bp65 and cyclooxygenase-2?COX-2?by CCl₄-induced.In summary,SPG?2.5 and 10 mg/kg?exhibited strong hepatoprective effect on the CCl₄-induced acute liver injury,which was related to its anti-oxidantive and anti-inflammatory capabilities.?2?Establishment of a D-GalN/LPS acute liver injury model.The D-GalN/LPS acute liver injury model was used to explore the pharmacological activity and mechanism of SPG protection of acute liver injury by D-GalN/LPS-induced.ICR mice were given SPG?2.5?10mg/kg?for 7 days by oral administration,and LPS?50?g/kg?and D-GalN?800 mg/kg?was intraperitoneally injected 1 hour after SPG administration on the last day.After 24 hours of D-GalN/LPS administration,the serum and liver of the mice were collected.Relevant indicators in serum and liver tissues were determined according to the kit instructions.The mechanism of SPG protecting D-GalN/LPS-induced acute liver injury was analyzed by H&E staining,Western-blot and RT-PCR.The results showed that compared with D-GalN/LPS model group,SPG significantly reduced the levels of AST,ALT,TTNF-?and IL-1?in serum,increased the levels of GSH,CAT and SOD,and decreased the levels of MDA in liver tissue.In addition,SPG could alleviate D-GalN/LPS-induced liver histopathological damage.Moreover,western-blot assay showed that SPG up-regulated the expression of heme oxygenase-1?HO-1?,nuclear factor E2 related factor 2?Nrf2?and Sirt1,and down-regulated NF-?Bp65 and I?B by D-GalN/LPS-induced.At the same time,RT-PCR results revealed that SPG down-regulated the levels of TNF-?and IL-1?mRNA by D-GalN/LPS-induced.In conclusion,SPG might?2.5 and 10 mg/kg?significantly protected D-GalN/LPS-induced acute liver injury,and its mechanism of action was related to its anti-oxidation and anti-inflammatory capabilities.?3?Establishment of an APAP acute liver injury model.The pharmacological activity of SPG-protected APAP-induced acute liver injury was explored by using APAP acute liver injury mouse model.ICR mice were given SPG?2.5?10 mg/kg?for 7 days by oral administration,and APAP?300 mg/kg?was intraperitoneally injected 1 hour after SPG administration on the last day.After 24 hours of APAP administration,the serum and liver of the mice were collected.The serum levels of ALT and AST were determined according to the kit instructions.The results showed that there was no significant difference in the serum ALT and AST levels between the SPG pretreatment group and the APAP model group.Therefore,SPG?2.5 and 10 mg/kg?did not have a significant protective effect on APAP-induced acute liver injury.