| Panax ginseng is a kind of precious Chinese herbal plant,which has high medicinal value and is widely used in medical and health care industries.It is found that the main active component of ginseng is ginsenoside.There are many kinds of ginsenosides in ginseng.Different ginsenosides have different pharmacological activities,especially some rare ginsenosides have better pharmacological activities than common natural ginsenosides.However,due to the small content in natural ginsenosides,it is difficult to prepare them in large quantities.Therefore,the transformation method is adopted It has become a developing trend to obtain rare saponins by means of method.Ginsenoside Re,as one of the most common saponins,has high content in ginseng and is easy to obtain.In this paper,the enzymatic transformation of ginsenoside Re provides a new way to enrich and prepare rare ginsenoside with high pharmacological activity.Objective: on the basis of laboratory research,ginsenoside Re was degraded by enzymatic method,and ginsenoside Rg1 and Rh1 were purified,and their pharmacological activities in anti-inflammatory,APAP induced acute liver injury and Alzheimer's disease activity were studied.Methods:(1)preparation of Ginsenoside Rg1 and Rh1: ginsenoside Re was transformed by?-glucosidase method.The degradation products of ginsenoside Re were separated,purified and detected by AB-8 macroporous resin method,silica gel column chromatography,thin layer chromatography and high performance liquid chromatography.(2)To study the activity of AD(Alzheimer's disease): to establish AD mice model,to study the effect of ginsenoside Rh1 on cognitive impairment of AD mice model by dark avoidance test,platform jumping test,Morris water maze test and acetylcholine(Ach)kit.(3)the anti-inflammatory activity of research: first of all,inflammation mice model was established,and then through the mouse ear edema induced by xylene and carrageen glue induced paw edema in mice experiment,acetic acid in mice,the influence of blood capillary permeability of the abdominal and body torsion test,using the kits serum inflammatory cytokines in mice tumor necrosis factor(TNF-?),interleukin 6(IL-6),interleukin-1 the expression level of beta(IL-1?),as indicator,explore Rg1 and ginsenosides Rh1 anti-inflammatory activity in the body.(4)study on the activity of acute liver injury caused by APAP: Acetaminophen(APAP)is adopted to establish the mice liver injury model,through the kits in mice serum tumor necrosis factor(TNF-?),interleukin 1 beta(IL-1 ?),interleukin 6(IL-6)and superoxide dismutase(SOD),aspertate aminotransferase(GOT),malondialdehyde(MDA)and the expression level of cereal third transaminase(GPT),observe the liver tissue pathological changes,immune protein analysis of Bax,Bcl-2 protein detection,research of Rg1 ginsenosides and Rh1 APAP cause liver injury protection.(3)ginsenoside Rg1 and Rh1 can play an anti-inflammatory role by regulating the expression of inflammatory factors.Results:(1)ginsenoside Re was degraded to monomer ginsenoside Rg1 by enzymolysis,with the conversion rate of 59.25% and extraction rate of 11.21%.The conversion rate was67.75% and the extraction rate was 11.73%.(2)Ginsenoside Rh1 can improve cognitive impairment in AD mice.Compared with the negative group,saponin Rh1 can increase the expression of Ach in the hippocampus of mice.In the dark avoidance experiment,the incubation period of the saponin Rh1 dose group increased relatively,the incubation period of the Rh1low-dose group was 144.5±29.3s(P <0.05),and the incubation period of the Rh1 medium-dose group was 150.3±18.2s(P<0.01),The incubation period of the Rh1 high-dose group regression test was 132.7±22.6 s;in the platform jumping test,the incubation period of each dose group of saponin Rh1 increased to varying degrees,the incubation period of the Rh1 low-dose group was111.3±15.8s,and the incubation period of the Rh1 medium-dose group was 132.6±36.7s(P<0.01),the incubation period of the high-dose Rh1 group was 102.9±23.3s;in the Morris water maze test,the swimming path of each saponin Rh1 dose group was simpler,and the time to find the platform was relatively shorter.(3)Ginsenosides Rg1 and Rh1 can play an anti-inflammatory role by regulating the expression levels of inflammatory factors.Saponins Rg1 and Rh1 significantly inhibited the expression of inflammatory factors TNF-?,IL-6,IL-1?;can alleviate the paw swelling symptoms in mice,the paw swelling inhibition rate in the Rg1 high-dose group was 41.8%,and the paw swelling inhibition in the Rh1 medium-dose group The rate is 33.5%;it can relieve ear swelling symptoms,the ear swelling inhibition rate in the Rg1 high-dose group is57.2%,and the ear swelling inhibition rate in the Rh1 medium-dose group is 80.9%;and the effect of acetic acid-induced abdominal capillary permeability in mice The permeability inhibition rate of Rg1 high-dose group was 40.7%,and the permeability inhibition rate of Rh1medium-dose group was 47.7%;and the number of writhing in mice was also significantly reduced.The inhibition rate of writhing response in Rg1 high-dose group was 43.8%.The inhibition rate of writhing response in the middle-dose Rh1 group was 41.5%.(4)Ginsenoside Rg1 and Rh1 have the protective effect of improving the liver injury caused by APAP.Compared with the negative group,the content of MDA in the serum of the mice in each dose group of Rg1 and Rh1 decreased,the activity of GSH and SOD increased significantly,and the content of GOT,GPT,TNF-?,IL-6 and IL-1?in the serum decreased.HE staining showed thatthe necrosis area of liver tissue decreased significantly.The results of immunoprotein showed that Bax protein decreased and bcl-2 protein increased.Conclusion: ginsenoside Re can be degraded to Rg1 and Rh1;Ginsenoside Rh1 can improve the cognitive impairment of AD mice;Ginsenoside Rg1 and Rh1 can inhibit the inflammatory response to a certain extent,and can effectively inhibit the liver injury caused by APAP by inhibiting oxidative stress,inflammatory response and hepatocyte apoptosis. |
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