| [Objective] Explore the clinical and experimental study of BXD in improving metabolic inflammatory injury of T2 DM in order to increase the application range of BXD;at the same time,the NP technology was used to analyze the action target and related signal pathway of BXD in the treatment of T2 DM,and to further analyze the theoretical basis and mechanism of its prevention and treatment of T2 DM.Through clinical and basic trials,to verify the effect of TCM in regulating energy metabolism pathway,restoring the balance of glucose?lipiding metabolism and improving IR.[Method] Network Pharmacology: the target genes of BXD were obtained by using the database of traditional Chinese medicine system pharmacology analysis platform(bioinformatics analysis tool for molecular mechanism of TCM,BATMAN-TCM),and the possible fractions of BXD were used as targets.The target genes of T2 DM were collected from OMIM and TIDDI disease databases.After the genes were intersected,the protein-protein interaction network was constructed by STRING.Through the R plug-in biological information analysis tool for GO enrichment analysis and the KEGG sub-signal pathway.Above of all the results were displayed visually.Clinical Study: Using the method of randomized controlle study,cases of T2 DM with spleen deficiency and stomach stagnation were collected from the inpatient department and outpatient department which Hospital of Chengdu University of tcm from October 2018-October 2019.According to the principle of random grouping(random number table),they were divided into the control group and the treatment group.The control group was treated with the original hypoglycemic western medicine,and the treatment group was treated with BXD on the basis of western medicine.The course of treatment was 12 weeks.Basic Research: The animal model of T2 DM was established by the early mature method.80 male SD rats were randomly divided into normal group(NG,10 rats)and model group(DM,70 rats).50 rats were randomly divided into model group(DM),metformin group(MET,0.16g·kg-1),Banxia Xiexin Decoction dose group(BXD,6.98g·kg-1),NG and DM were respectively given the same volume of normal saline for 4 weeks.The contents of fasting blood glucose,TC and TG in serum were detected by biochemical methodinsulin.FFA,ADPN,TNF-? and IL-6 in serum and SOD,GSH Px,ATP?ATP / AMP in liver tissue were detected by ELISA.The contents of AMPK??SIRT1?PGC-1?and PPAR ? in liver tissue were detected by PCR and WB?[Results] Network pharmacology:Eleven of the 245 target genes of BXD are associated with T2 DM.The overlapping genes are PPARD,INS,TNF,SOD2,MTNR1 B,KCNB1,SIRT1,GPBAR1,ADORA1,GHRL and AVPR2.Clinical trial:A total of 56 cases were collected,including 3 cases of abscission and 3 cases of rejection;50 cases were actually completed,including 26 cases in the control group while the other cases in the treatment group.The baseline data of gender and age were the same in the two groups(P > 0.05).(1)After treatment,compared with the control group,the P values of FPG and P2 HPG were less than 0.05(P < 0.05),and the P values of Hb A1 c and HOMA-IR were less than 0.05(P < 0.05);(2)After treatment,compared with the COT the P values of TC and TG were less than 0.05(P < 0.05),the difference was statistically significant;(3)Compared with the COT,the concent of FFA and ADPN in the treatment group were significantly lower(P < 0.05),the difference was significant.(4)Compared with the COT,the content of IL-6 and TNF-? in the treatment group decreased significantly(P < 0.05).(5)After treatment,compared with the control group,the concentration of ATP in the treatment group decreased(P<0.05),the ratio of AMP/ATP decreased(P>0.05).while the concentrations of SIRT1,AMPK?and PGC-1? in the TRT were lower than those in the control group(P < 0.05). Basic Test 1.Establishment of T2 DM rat model and effect on blood glucose(1)Through general observation,compared with ng group,the rats in model group were not sensitive to activities,weight loss,dull fur,diet and other intake decreased;(2)In the model group,fasting blood glucose,insulin and insulin resistance index increased significantly(P<0.05);(3)After the intervention of BXD,the weight,mental state,fasting blood glucose,insulin and insulin resistance index of the model group were improved.The model of T2 DM rats was established successfully.2.Effect of BXD on blood lipid metabolism in DM Rats Compared with NG group,the content of TC,TG and FFA in DM group was significantly higher,while the content of ADPN was significantly lower(P<0.01).Compared with DM group,the content of TC,TG and FFA in drug intervention group was significantly lower,and the content of ADPN was significantly higher(P<0.01 or P< 0.05).The change of BXD group was more significant.3.Effect of BXD on inflammatory factors and oxidative stress in DM Rats Compared with NG,the serum IL-6 and TNF-?of DM rats increased significantly(P< 0.01),On the contrary in the DM.The liver GSH PX and SOD of DM rats decreased without statistical difference(P>0.05),compared with NG.GSH PX and SOD in liver tissue of rats in group BXD were significantly higher than those in group DM(P<0.05);4.Effect of BXD on the related regulatory factors of AMPK signal pathway in DM Rats Compared with NG group,the content of ATP and AMP/ATP in liver tissue of DM group was significantly lower(P<0.01);After drug intervention ATP and AMP/ATP increased.Compared with NG group,the content of SIRT1 m RNA in liver tissue of rats decreased significantly,while the content of PGC-1?m RNA decreased significantly,the content of SIRT1 m RNA,AMPK?and PPAR?m RNA in liver tissue of rats in drug intervention group increased significantly,compared with that in DM group,and there was significant difference between drug intervention group DM group.The content of AMPKa m RNA and PPAR? m RNA in liver tissue of rats in drug intervention group was significantly higher than that in DM group,and the content of AMPKa m RNA and PPARa m RNA was significantly higher.Compared with NG group,there was no significant difference in the expression of AMPK?and PPAR? protein in the liver tissue of DM group(P>0.05),and the expression of PGC-1? protein increased;The expression of SIRT1 protein decreased significantly(P<0.05);compared with DM group,the expression of PGC-1?protein in the liver tissue of drug intervention group was significant The decrease was statistically significant(P<0.05 or P<0.01).There was no significant difference in SIRT1 and PPAR?protein expression(P>0.05).[Conclusion](1)Through gene expression,BXD can regulate inflammatory factors,oxidative stress,energy metabolism,affect the binding of insulin and its receptor,induce SIRT1 expression and activate it AMPK/SIRT1/PGC-1?signaling pathway regulates the opening of energy metabolism,promoting glycolipid metabolism and mitochondrial biosynthesis to enhance insulin sensitivity.To explain the advantages of multi-target and multi-channel medicine in the prevention and treatment of T2 DM,and provide research direction and theoretical reference for the follow-up experimental research.(2)T2DM can cause disorder of glucose and lipid metabolism,inflammation,oxidative stress injury,and lead to imbalance of energy metabolism.In DM group,there are more inflammatory cell infiltration and steatosis in pancreatic tissue and liver tissue,and the ratio of lipid droplets in liver tissue is significantly increased.(3)BXD can lower blood sugar of type 2 diabetic patients and rats,lower insulin resistance index,improve insulin resistance,down-regulate the content of IL-6 and TNF-?,regulate the level of pro-inflammatory factors,and enhance the body's sensitivity to insulin;Reduce the content of ADPN,activate the AMPK system,increase the body's consumption of FFA,reduce the levels of TC and TG and insulin resistance,and thereby exert a lipid-lowering effect;meanwhile,BXD can upregulate AMP in type 2 diabetic patients and rats / ATP ratio,activate the AMPK/SIRT1/PGC-1? system,increase the ATP content,thereby promoting the body's gluconeogenesis,fatty acid oxidation and mitochondrial biosynthesis to enhance anti-inflammatory,antioxidant and regulate energy metabolism.(4)BXD can inhibit fatty acid oxidation and pro-inflammatory cytokines,decrease FFA content and SOD and GSH activity by activating PPAR ? downstream molecule of PGC-1 ? in DM rats,so as to reduce oxidative stress damage and lipid accumulation of rat hepatocytes,improve IR,and restore the level of glucose and lipid metabolism.combined(5)With the study of this experiment,we think that the key of "spleen does not dissipate essence" lies in the functional disorder of "spleen governing movement and transformation",in which "spleen governing transformation" may play a major role. |
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