Effects Of Danggui-Shaoyao-San And JianPi Lishi Recipe On Cognitive Impairment And The Changes Of Glycosylation Of Estrogen α Receptor O-GlcNAc In Female Db/db Mice

ObjiectiveEpidemiological studies have found that diabetes could cause Pathological changes in the neurophysiological structure of the brain,leading to cognitive dysfunction.More and more studies have found that Danggui Shaoyao San(DSS)can effectively relieve diabetes and cognitive dysfunction.The study aimed to investigate whether Danggui Shaoyao San(DSS)and Fuling Baizhu Zexie(FBZ)could improve the cognitive dysfunction of female diabetic mice by inhibiting the abnormal modification of O-GlcNAc glycosylation of ERa in db/db mice.Which can Provide new therapeutic targets for diabetic encephalopathy.MethodsThe study was used female spontaneous diabetes model db/db mice as a classic model for studying diabetes.The 12-week-old db/db female mice were randomly divided into the following six groups after being adaptively reared for one month:model group,DSS high-dose group,DSS low-dose group,FBZ high-dose group,FBZ low-dose group and Positive drug metformin(Met)Groups,with 12 in each group,and 12 homologous db/m female mice were used as the control group.The drugs in each group were given by intragastric administration for 10 weeks.The normal control group and the model group were replaced with 0.9%saline.After the gavage,Oral Glucose tolerance test and Insulin tolerance test were used measure the changes of glucose tolerance and insulin tolerance in db/db mice;Morris water maze was used to measure the changes in the sPatial learning and memory abilities in db/db mice;The blood biochemical test was used to measure blood glucose and blood lipid levels in db/db mice;Nissl staining was used to measure the changes of brain neurons in db/db mice;ELISA kits(PGE2,TXB2 and LTB4)were used to measure the levels of inflammatory mediators in db/db mice;MDA,SOD,CAT and GSH-PX kits were used to detect the levels of oxidation Stress in db/db mice;NO,iNOS and TNOS kits were used to detect the levels of nitriding stress in db/db mice;LDH,PK and HK kits were used to detect the levels of glucose metabolism in db/db mice;Western blot was used to detect the expression of O-GlcNAc glycosylation,O-linked N-acetylglucosaminidase(OGT),O-linked N-acetylglucosaminidase(OGA)and estrogen recePtor ERa in db/db mice brain.In addition,immunofluorescence was used to detect the intensity and co-localization of estrogen receptor ERa and O-GlcNAc glycosylation in db/db mice brain.ResultsThe results of Morris water maze showed that compared with the normal control group,the escape latency of the model group was significantly longer,the DSS group’s escape latency was shortened,and the swimming trajectory was relatively simple and clear.Although the FBZ group could reduce the escape latency,but the difference is not statistically significant.DSS could improve the learning and memory ability of female db/db mice,especially the high-dose DSS group;Nissl staining results showed that DSS could reduce hippocampal neuron damage in db/db mice.Oral Glucose tolerance test results showed that blood glucose and the area under the curve were decreased,which indicated that DSS could reduce impaired glucose tolerance in db/db mice.Insulin tolerance test results showed that the area under the curve was not decreased,indicated that DSS and FBZ could not improve the impaired insulin function of db/db mice.The blood biochemical test results showed that both DSS and FBZ could improve the hyperlipidemia and high blood glucose of db/db mice.The activities of SOD,GSH-PX and CAT were increased,and the levels of MDA,NO,iNOs and TNOS were decreased,indicated that the DSS and FBZ could alleviate oxidative stress and nitriding stress damage in db/db mice,while the effect of DSS was better than FBZ.The levels of PK,HK and LDH were decreased,indicated that DSS and FBZ could reduce the level of glucose metabolism in db/db mice brain.The levels of PGE2,TXB2 and LTB4 were decreased,indicated that DSS and FBZ could alleviate inflammatory damage in db/db mice brain.Western blot(ERα,O-GlcNAc,OGA and OGT)and immunofluorescence(ERa and O-GlcNAc)results showed that the expression of ERa and OGA were increased,while the expression of O-GlcNAc and OGT were decreased,indicated that DSS and FBZ could increase the content of estrogen receptor ERa in db/db mice brain,and reduce the level of O-GlcNAc glycosylation in db/db mice brain.ConclusionsDSS and FBZ could improve the cognitive impairment of diabetic mice by influencing glucose and lipid metabolism disorder,regulating glucose metabolism level,reducing oxidative stress and inflammation damage;and the mechanism was related to inhibit the abnormal modification of O-GlcNAc glycosylation of ERα.