Total Saponins From Anemarrhena Asphodeloides Improve Cognitiverole Of Diabetes And Depression Model Rats

ObjectiveDepression and diabetes are closely associated with cognitive impairment,and can increase the risk of developing Alzheimer' s disease,clinical diabetes is often associated with depression,which can increase the severity of cognitive impairment.Modern pharmacological studies have shown that Total saponins from anemarrhena asphodeloides has improves diabetes,antidepressant,improves cognitive impairment and other effects,the main saponins,about 6%.This subject from the perspective of anti-apoptosis hypothesis of anemarrhena asphodeloides treat depression,diabetes-related mechanisms of cognitive impairment.Methods1.Model establishmentDiabetes with depression model rat of established:100 male SD rat,according to weight and the blood sugar lipid,used half random of method is divided into 9 group,10 each group,named as normal group(NC group),depression model group(MD group),MD administration group(ZMD);12 each group,diabetes named as model group(DM group),composite model group(DD group),DM administration group(ZDM group),DD administration high dose group(ZDD-H),DD administration low dose group(ZDD-L),DD positive drug group(DD-P).Used chronic unpredictive stimulus(CUMS)induced depression model,STZ and high fat feed induced 2 type diabetes model,CUMS+STZ+high fat induced composite model,through continuous irrigation stomach with Total saponins from anemarrhena asphodeloides for 17 week.2.from the angle of anti-apoptosis,explore the mechanism of anemarrhena asphodeloides to treat cognitive impairment of diabetes mellitus and depressive.Check the Group rat weight every week.Administration 12 weeks,biochemical method detection plasma FPG,TC,A?1-42,GLU,and BDNF.Administration 12 weeks,Open-Field experiment,and Barnes maze experiment evaluation behavior learn,to drug 17 week,Anatomy save rat brain organization,HE staining of pathologic Changes in hippocampus of rats,Immunohistochemistry assay in hippocampus of rats in each group the expression protein of NR2A?NR2B?P-JNK?BAX and BCL-2,Caspase-3 and real-time fluorescence quantitative PCR detection the expression of NR2A mRNA?NR2B mRNA?JNK mRNA?BCL-2 mRNA and BAX mRNA Caspase-3 mRNA in hippocampus of rats in each group.Results1.Evaluation of external indicatorsBecause of depressive model,the weight of MD Group and DD Group were significantly lower than the normal group.Because of diabetes model,DD Group of rats body weight can be higher than MD Group,DM group due to the amount of stz abdominal injection,weight and normal group did not significantly difference,the administration group rat of depression model,weight did not change significantly,but the diabetes model group after the drug significantly decreased.In Open-field experiment,depressive model rats decreased significantly,and depressive behavioral characteristics were obvious,and the depressive behavioral state of rats was significantly improved by anemarrhena asphodeloides.In the experiment of Barnes Maze,the cognitive and spatial memory abilities of the rats were decreased markedly compared with the normal group in each model group,and the learning and memory ability of each drug group was improved in comparison with each model group,in which the effect of high dose group was slightly obvious than the low dose group.2.The indexes of blood and tissuesAfter 6?12?17 weeks of blood glucose test,compared with the normal group,MD group Rat blood glucose is slightly lower than the normal group,after the drug is not afraid of significant differences;compared with normal group,the fasting blood glucose(FPG)and total cholesterol(TC)of DM group of Rats,significantly increased(P<0.01),compared with normal group,DD Group rats have elevated blood glucose trend,but the trend is lower than the DM group;compared with the DM group,the total saponin can inhibit the increase of FPG and TC in ZDM Group.After 12 weeks,compared with normal group,the plasma A?1-42 protein and glutamic acid in the rat MD and DM Group was significantly elevated(P<0.05 or P<0.01),and the compound model group A?1-42 protein,glutamate did not have significant statistical significance;compared with MD and DM Group,the total saponins can significantly decrease the plasma A?1-42 protein and glutamic acid level in ZMD and ZDM group rats(P<0.05 or P<0.01),although DD Group was not statistically significant compared to normal group,the anemarrhena asphodeloides was reduced glutamate level(P<0.05 or P<0.01);Compared with the normal group,BDNF in the three rat model group decreased significantly(P<0.01),but with the anemarrhena asphodeloides,the BDNF did not increase,the difference was not statistically significant.After 17 weeks,the MD,DM,DD rat Hippocampus A ? 1-42 protein and glutamic acid increased significantly(P<0.01),compared with two separate model groups,DD rat A ?1-42 protein and glutamic acid level,a slightly higher,there is a certain model superposition trend,but not statistically significant;compared with the corresponding model group,the total saponin can significantly reduce the level of A?1-42 protein and glutamic acid in the hippocampus of rats(P<0.05 or P<0.01)By HE staining,the neurons in the hippocampus CA1 area of rats were observed,and the cells were intact and dense in the normal group of rat hippocampus.The neurons arranged neatly,coloring evenly,morphological rules,the nucleus of large and round;MD,DM,DD Model group of rat hippocampal neurons decreased,a large number of nuclear contraction,the gap between the cells deepened,the emergence of some irregular neurons,the necrosis of the small glial cells increase.3.Through NR-JNK-BCL-2,Bax,Caspase-3 pathway to explore the role of anemarrhena asphodeloides improving cognitive impairment.In this study,we test the average light density of rat Hippocampus CA1 region NR2A,NR2B,P-JNK,BCL-2,BAX and Caspase-3 proteins.The results showed that three model group NR2A,NR2B,P-JNK,Bax light density increased(P<0.05 or P<0.01),compared with normal group,and Caspase-3 had no statistically significance;After the total saponin was given to the drug,the model group NR2A,NR2B,P-JNK,Bax of the light density decreased(P<0.05 or P<0.01).By real-time fluorescence quantitative PCR assay,the results showed that The expression of three model group NR2A mRNA NR2B mRNA,JNK mRNA,Bax mRNA was a significant increase or in the trend,BCL-2 mRNA expression decreased(P<0.05 or P<0.01),Caspase-3 mRNA expression was not statistically significant,compared to normal group.;After the total saponin was given,the model group NR2A mRNA,NR2B mRNA,JNK mRNA and Bax mRNA expression decreased or in the trend,BCL-2 mRNA expression increased(P<0.05 or P<0.01).Conclusion1.Body weight results show that anemarrhena asphodeloides can reduce the diabetes model of rat body weight,the Depression model has no effect on the body when abnormal fat accumulation,know that total saponins can play its role in reduceding lipid.anemarrhena asphodeloides has the effect of reducing the accumulation of glutamate,decreasing the behavior of depressive rats,and reducing the effect of fasting blood glucose,total cholesterol,and improving blood glucose tolerance to cure diabetes.2.Through the experiment,the total saponin has A?1-42 protein in the plasma and hippocampus of the diabetic model and depressive model.Glutamate content,and improve the learning and memory ability of rats,regulating NMDA NR-JNK-BCL-2,Bax induced apoptosis pathway,indicating that diabetes mellitus and depression have similar cognitive impairment mechanisms,knowing that total saponins by regulating this pathway to improve the role of cognitive impairment.The anemarrhena asphodeloides of total saponin has no significant effect on BDNF,which may have no obvious regulating effect on the results of Caspase-3.