| Diabetes is a global epidemic disease.Its morbidity and mortality are subject to a yearly progressive increase.Therefore,the study of diabetes has always been a significant topic all over the world.Sutherlandia Frutescens(S.fru)is a Southern African legume which has traditionally been used as an indigenous medicine for a variety of ailments.Likewise,Leonurus Artemisia,as a commonly used medicine in China,also has extensive medical effect.Accordingly,the therapeutic effect of these two drugs on diabetes has become a well-concerning focus.This thesis is conducted using the extract of Leonurus Artemisia and Sutherlandia Frutescens as material basis as well as modern science and technology as method,establishing the type II diabetes zebrafish model,to probe into the toxic effect of leonurine,extract from Leonurus Artemisia,ethanol extract from Sutherlandia Frutescens(S.fru-OH)and water extract from Sutherlandia Frutescens(S.fru-H2O),and Standard D-pinitol on zebrafish,and analyze the impact and mechanism of these drugs on the glucose and lipid metabolism of zebrafish.To conduct toxicity study on zebrafish,we first divide subjects into several groups,including normal control group,leonurine group,ethanol extract from Sutherlandia Frutescens group,water extract from Sutherlandia Frutescens group,and D-pinitol positive control group.Zebrafish we select as object are fertilized within 24 hours and proceed normal development.We medicate them and observe the changes of their morphological characteristics,cardiac development,aberration rate,death rate,hatchability and so on.It turns out that when the concentration of S.fru-OH reaches 30μg/m,juvenile fish all die in the experimental period,while the same death condition would only occur when the concentration of S.fru-H2O reaches 200 μg/m.When the concentration of leonurine reaches over 100 μg/m,zebrafish would experience sever pericardial cyst and cyst vitelline,but the death rate is lower than that medicated with the two extracts of S.fru.When the zebrafish are medicated with D-pinitol,rapid mass death condition only occurs when its concentration reaches 300 μg/m,and the aberration rate is relatively low at any concentration.This toxicity test demonstrates that the lethal toxicity and teratogenic toxicity of S.fru-OH are stronger than those of S.fru-H2O,and the effect of D-pinitol is close to that of S.fru-H2O.Moreover,high concentration of leonurine has strong teratogenic toxicity,but would not necessarily cause death,and toxicity would not show up at a low concentration.These conclusions provide reference of concentration for the following experiments.Finally,we medicate zebrafish fertilized within 96 hours with 100 μM cAMP and 1mM DEX.After incubation for 48 hours,we establish type Ⅱ diabetes zebrafish model by PEPCK gene overexpression.We then conducted mRNA expression quantity test,by detection of RT-qPRC,on relevant genes involved with insulin and glycometabolism of the zebrafish of PEPCK gene overexpression.Those relevant genes include PEPCK,GLU2,GP,GYS1,G6Pase,PFKFB3,and so on.It turns out that after zebrafish of type Ⅱ diabetes model are treated with the two extracts of S.fru and leonurine,expression quantity of multiple target genes including EPCK,GLU2,GP,GYS1,G6Pase,PFKFB3 would notably increase or decrease.It demonstrates that extracts of S.fru and leonurine could affect glycometabolism by regulating some genes involved with glucolipid metabolism,and thus improve insulin resistance.In terms of reducing blood sugar,S.fru-OH performs better than S.fru-H2O,and the effect of leonurine is also passable.In conclusion,leonurine,S.fru-OH,and S.fru-H2O have positive hypoglycemic effect on zebrafish of type Ⅱ diabetes model.This thesis makes a further exploration on the anti-diabetic mechanism of these drugs,and thus offers new ideas and lays a foundation for research and development of anti-diabetic drugs. |
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