Effects Of Atractylodes Macrocephala Koidz Combined With Cuscuta Chinensis Lam On Glycolipid Metabolism And The Expression Of Pi3k And Akt In Drosophila

Objectives Atractylodes Macrocephala Koidz combined with Cuscuta Chinensis Lam（AMK-CCL）was used to intervene the insulin-resistant diabetic Drosophila model,and observe its effects on the glucose and lipid metabolism of Drosophila diabetic model,as well as investigate the preliminarily mechanisms.Methods Using the GAL4/UAS dual expression system and the genetic hybridization assay,we specifically impeded the physiological function of the Drosophila Insulin Receptor（In R）in the fat body,reduced the transduction activity of the Insulin signaling pathway to causes Insulin Resistance（IR）,thereby establishing a Drosophila IR diabetes model.Three strains of w¹¹¹⁸ male flies,Cg-GAL4 female flies and UAS-In R^K1409A male flies were selected to cross at the ratio of 10:6,that is,10 female virgin flies and 6 male flies were randomly selected into a hybrid tube.There are 10 parallel tubes for each group.In control group,Cg-GAL4 female virgin flies were crossed with w¹¹¹⁸ male flies,and the other groups were Cg-GAL4 female virgin flies with UAS-In R^K1409A male flies.They were randomly divided into control group,model group,metformin group,and low,medium and high dose groups that adding the alcohol extract of AMK-CCL,and the middle dose is further divided into two groups.The control group and the model group were reared in normal fruit fly culture medium,and the other groups were kept in the Drosophila culture medium,which contains 0.00165g/m L metformin,0.0125g/m L,0.025g/m L,0.05g/m L and0.1g/m L AMK-CCL alcohol extract.Observe the growth status of each group,and collect the Cg-GAL4/+third-instar larvae as the control group samples,and the Cg-GAL4/+;UAS-In R^K1409A/+third-instar larvae as the remaining samples group samples.All the above samples were tested for glucose,trehalose,glycogen,triglyceride,and protein concentration,and the above in vivo indexes were calculated to obtain the optimal concentration of the AMK-CCL.Then,the groups were divided into the control group,the model group,the metformin group,and the optimal dose group of AMK-CCL.The reporter gene of Phosphatidylinositol 3 kinase（PI3K）activity-t GPH was introduced into the IR model to determine the PI3K status;Western blot method was used to detect the expression of Protein kinase B（Protein kinase B,Akt）and p-Akt protein in the fat body of Drosophila.Finally,green fluorescent protein（GFP）was inserted into the Drosophila Forkhead box protein O（d Fox O）gene region（d Fox O-GFP）to marker the endogenous intracellular localization and expression of d Fox O in fatbody cells,and thus monitor the PI3K activity,phosphorylation level of Akt,and transcription activity of d Fox O in the above four groups.Results 1 After grouping and crossing,compared with the control group,the model group displayed fewer progeny reproduction,delayed development,reduced activity,smaller body size,and weight loss;compared with the model group,the AMK-CCL group had the above mentioned Drosophila.The situation has been improved significantly.2Compared with the control group,the circulating glucose and circulating trehalose levels in the model group increased obviously,and the content of protein,glycogen/protein,or triglyceride/protein decreased largely,indicating that the energy storage capacity of the Drosophila fat body declines,P<0.05.3 Compared with the model group,the levels of circulating glucose in the AMK-CCL treated group decreased,and the low dose of this formula was the best dose,P<0.05.4 Compared with the normal group,the protein content in model group reduced,the PI3K activity and the phosphorylation level of Akt reduced significantly（P<0.05）,while the transcriptional activity of d Fox O was upregulated;compared with the IR model group,the activity of PI3K and phosphorylation level of Akt increased significantly（P<0.05）,while the transcription activity of d Fox O reduced largely in the AMK-CCL group.Conclusions 1 The optimal intervention dose of AMK-CCL for insulin-resistant diabetic Drosophila model is 0.0125g/m L.2 AMK-CCL reduces the circulating glucose concentration in IR diabetic Drosophila model.3 AMK-CCL improve the energy storage capacity of glycogen and triglycerides,and increase the protein content in Drosophila.4AMK-CCL may ameliorate insulin resistance in the fat body of diabetic Drosophila,by which that improving the activity of PI3K and the phosphorylation level of Akt,and inhibiting the transcription activity of d Fox O.Figure 23;Table 7;Reference 108...