Study On The Quality Analysis And Pharmacological Activity Of Total Saponins Of Ginseng Stem And Leaf And Flavored Black Ginseng

In this paper,the thesis focused on the research ginseng stems and leaves of total saponins（SLG）and garlic as processing aid for ginseng get new ginseng products-flavored black ginseng（FBG）.The objective of this project was to investigate the quality and biological activity of SLG and FBG,and the main research contents and results are as follows:1.Quality analysis of total saponins of ginseng stems and leaves（SLG）and flavor black ginseg（FBG）.Firstly,the content of 20 ginsenosides in SLG and FBG was determined by HPLC,and the contents of total sugar,reducing sugar,protein and 5-hydroxymethylfurfural in FBG were determined by phenol sulfuric acid methods,two nitro salicylic acid methods（DNS）,Coomassie Brilliant Blue methods and HPLC,respectively.Finally,study on the transformation of ginsenosides in FBG by garlic.The results showed that SLG contains 20 kinds of ginsenoside content of a total of 69.99%,and common ginsenosides were converted into rare saponins such as ginsenoside Rk3,Rg5 and so on in FBG.Meanwhile,reducing sugar accounted for 92.06%of the total sugar content and the protein content was very low,and the content of 5-HMF was as low as 7.40 mg/kg in FBG.It was found that garlic can promote ginsenoside conversion,with the longer the processing time,the higher the temperature,the less polar ginsenoside conversion when studying the effect of garlic from processing time and temperature.2.Study on preventing obesity effect and its mechanism of action of SLG.The study of SLG on the prevention of obesity was conducted by dividing the groups into experimental animal models with administration with high fat diet（HFD）.At the end of the experiment,serum and organ tissues were taken,and the index of organ was calculated;the relevant indexes in serum were determined according to the instructions of the kit;and the content of acylcarnitine in the muscle tissue was determined by mass spectrometry.At last,H&E staining,immunohistochemistry,Western blot and RT-PCR were used to analyze the liver and adipose tissue.The results showed that SLG significantly reduced weight gain and decreased organ index in C57BL/6J obese mice induced by HFD.Compared with HFD,SLG reduced free fatty acid（FFA）,Leptin,Glucose and Insulin levels,relieved dyslipidemia,and regulated acylcarnitine levels.In addition,SLG could down-regulated the expression of PPARγ,FAS,CD36 and FATP2,and up-regulated the expression of CPT-1,UCP-2,PPARα,HSL and ATGL in liver tissues.Meanwhile,compared with HFD group,SLG group reduced mRNA levels of PPARγ,aP2 and Leptin and up-regulated the gene expression of PPARα,PGC-1α,UCP-1 and UCP-3 in adipose tissue.It can be seen that SLG significant effect of preventing obesity in the range of 125-500mg/kg,and its mechanism may be related to promoteβ-oxidation and thermogenesis of mitochondrial,inhibit lipogenesis,promote lipolysis and reduce the intake of free fatty acids.3.Study on anti-tumor effect and its mechanism of action of FBG and FBGP.Study the anti-tumor effect of FBG and its polysaccharide by the establishment of H₂₂ tumor-bearing mouse model.After the experiment,the serum,organ and tumor tissues were taken,and the tumor inhibition rate and organ index were calculated;the indexes of serum physiological and cytokines were determined according to the instructions of the kit.Finally,tumor tissue detected by H&E staining,TUNEL staining,immunohistochemistry,Western blot and qRT-PCR methods.The results showed that the high dose of FBG and FBGP was 61.74%and 57.84%,respectively,and could increase immune organ index and cytokine content with no obvious side effects.In addition,FBG and FBGP could induce tumor cell apoptosis,the expression of up-regulate Bax and down-regulate Bcl-2 and VEGF by H&E staining,TUNEL staining,immunohistochemistry,Western blot and RT-PCR analysis.Therefore,FBG and FBGP could exhibit an inhibitory effect on H₂₂ tumor-bearing mice,which mechanism may be related to improving immune function,inhibiting angiogenesis,and inducing tumor cell apoptosis.4.Study on anti-depressant effect and its preliminary mechanism of action of FBG.At first,tail suspension test（TST）and forced swimming test（FST）of the classical behavior desperation model were used to investigate the antidepressant effects of FBG and the best dose was screened out.Next,study serotonergic,norepinephrine,dopaminergic,GABAergic and glutamatergic systems was involved in the antidepressant effect of FBG in FST by receptor antagonist or agonist pretreatment.Finally,the levels of 5-HT,NA,DA,GABA and GLU in the hippocampus of mouse brain were measured.The results showed that compared with the model group,FBG（100 mg/kg）significantly shortened the immobility time of mice in TST and FST,but had no effect on spontaneous activity test.In addition,FBG recreased levels of 5-HT,NA and GABA,decrease level of GLU level and no significant change level of DA by the blockade experiment and the measurement of brain tissue homogenate.It is concluded that FBG has obvious antidepressant effect,and its mechanism may be mediated by serotonergic,noradrenergic,GABAergic and glutamatergic systems.