Regulatory Mechanism Of Salvianolic Acid B On Metabolic Disorders In Mice With Diabetic Nephropathy

BackgroundDiabetic nephropathy is one of the most serious microvascular complications of diabetes,it is the leading cause of glomerular sclerosis and end-stage renal disease(ESRD)worldwide.Its incidence rate is increasing year by year,bringing heavy mental and economic burden to patients and their families.Nowadays,how to effectively control the incidence of DN has become a difficult problem in the medical field.The pathogenesis involves multiple pathways and many targets,and the effects of glucose and lipid metabolism disorders on diabetes and its complications are particularly important.Under the leadership of the instructor Professor Luo Ren,the research team used the evidence-based medical research to find the treatment of stage III of diabetic nephropathy with "Yiqi Yangyin Huoxue"therapy.Combined with the clinical experience of the instructor for many years,the Chinese medicine compound Qidan Dihuang Granule was developed(NO.ZL201310015464.5).Multi-center randomized controlled clinical trial results show that Qidan Dihuang Granule can significantly reduce the proteinuria of stage III of diabetic nephropathy,preliminary work confirmed that salvianolic acid B is one of its possible important active ingredients;It is one of the important pathological foundations of diabetic nephropathy,and salvianolic acid B has a significant effect on the symptoms of diabetic nephropathy.This study based on metabolomics technology to propose a scientific hypothesis:salvianolic acid B may regulate metabolic disorders in the body to achieve the role of treatment of diabetic nephropathy.Therefore,this subject was treated with salvianolic acid B in db/db mice with diabetic nephropathy model.Metabolomics was used to analyze the chemical constituents and metabolic pathways of salvianolic acid B absorbed into blood by DN mice.It provided more evidence for the treatment and improvement of diabetes and its complications.MethodEighteen 8-week-old male db/db mice with C57BLKs/J background were used in the experiment;and 6 male healthy db/+mice of the same background and same age,according to blood glucose,food intake,body weight,and water intake.Animals were divided into control group,model control group,alogliptin group(Alogliptin)and salvianolic acid B group(Salvinolic acid).6 animals per group.The body weight of the mice,the amount of water and food intake,and all animals in the experimental group were measured for fasting body weight after administration.During the experiment,the food intake and water intake of the mice were recorded.Blood glucose and body weight were measured at 0,4,8,12,and 16 weeks,respectively.At the 16th week after the administration,the mice were sacrificed to collect blood lipid,liver function,renal function and myocardial enzyme related indicators.Analysis of the chemical constituents of salvianolic acid B absorbed into blood by DN mice by metabolomic techniques using ultra performance liquid chromatography quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS).Metabolic pathways were analyzed by multivariate statistical methods to find potential biomarkers to explore the regulation of salvianolic acid B on metabolic disorders of diabetic nephropathy,and to clarify the regulation mechanism of salvianolic acid B on metabolic disorders of diabetic nephropathy.Result1.Salvianolic acid B improved the food intake,water intake,body weight and blood glucose,blood lipids and myocardial enzymes in mice with diabetic nephropathy;the improvement of body weight,blood glucose and blood lipids by salvianolic acid B was close to that of alogliptin.Salvianolic acid B is superior to alogliptin in improving food intake,water intake and myocardial enzymes.2.Compared with the normal group,the model group mice were identified 23 significant differential metabolites by analyzing serum metabolites.In the model group,the expression of Glyceollin Ⅲ,PC(o-18:0/22:0),TG(20):1(11Z)/22:0/20;5(5Z,8Z,11Z,14Z,17Z)),QYNAD,3,4-Methylenesebacic acid,7-Hydroxytrichodermol,(S)-Angelicain,Auxin a,Sphingosine 1-phosphate were low;the expression of Vaccenic acid,Alpha-dimorphecolic acid,Docosa-4,7,10,13,16-pentaenoic acid,4-HdoHE,Taurohyocholate,Isotetrandrine,PC(16:1(9Z)/22:6(4Z,7Z,10Z,13Z,16Z,19Z)),PC(18:3(6Z,9Z,12Z)/22:6(4Z,7Z,10Z,13Z,16Z,19Z)),PI(16:0/22:4(10Z,13Z),16Z,19Z)),Ganglioside GA2(d18:1/18:0)were high.By analyzing these differential compounds,we found that calcium-dependent signaling pathways,choline metabolism regulation,ERS,Golgi-to-ER signaling pathway,G-protein coupled receptor signaling,sphingolipid metabolism-related pathways,lipids and lipoprotein metabolism pathways were involved.3.Compared with the model group,the salvianolic acid B group were identified 25 significant differential metabolites by analyzing serum metabolites.Salvianolic acid B increased the content of lysoPC(28:0),15-Acetyl-4-dexynivalenol,TG.(18:1(11Z)/24:0/20:5(5Z,8Z,11Z,14Z,17Z)),p-Cresol sulfate,Mequitazine,LysoPE(0:0/18:1(9Z)),LysoPE(0:0/20:1(11Z)),1-Arachidonoylglycerophosphoin-ositol,3-O-Sulfogalactosylceramide(d18:1/12:0),while reducing the content of SM(d16:1/24:1)(15Z)),Glyceollin Ⅲ,PC(o-16:0/22:0),Bovinic acid,Alpha-dimorphecolic acid,Eicosapentaenoic acid,Arachidonicacid,Docosa-4,7,10,13,16-pentaenoic acid,LysoPE(18:0/0:0),PA(20:4(5Z,8Z,11Z,14Z)e/2:0).By analyzing these differential compounds,we found that alpha-linolenic acid and linoleic acid metabolic pathways,polyunsaturated fatty acid synthesis,RORA,YAP1 and TAZ gene expression,mitochondrial biosynthesis,circadian clock regulation,ECBs signaling,SREBP,PLCβ,PPAR-α,PLA2 activates lipid metabolism pathways,lipids and lipoprotein metabolism pathways were involved.Conclusion1.Salvianolic acid B was similar to alogliptin in terms of therapeutic effects,including hypoglycemic and lipid-lowering effects in diabetic nephropathy db/db mice;At the same time,it can also improve the symptoms of polydipsia in db/db mice and improve the quality of life in mice2.db/db mice had significant differential metabolites with normal control mice.These metabolites were mainly involved in lipid and lipoprotein metabolism,biosynthesis of unsaturated fatty acids,arachidonic acid metabolism,sphingolipid metabolism,and acetylcholine.Important metabolic pathways such as synthesis,fatty acid,triacylglycerol and ketone metabolism,glycolipid metabolism disorder;Salvianolic acid B regulated the metabolic disorder of the body by regulating these metabolic pathways to achieve the treatment of diabetic nephropathy...