Screening of Differentially Expressed Proteins in Skeletal Muscle of Type 2 Diabetes KKAy Mice Treated with Couplet Medicines of Scutellariae Radix and Coptidis Rhizoma Based on nano-LC-LTQ/Orbitrap-MS
LIU Tong-tong, XIAO Qin, SHENG Jun-qing, et al. Screening of Differentially Expressed Proteins in Skeletal Muscle of Type 2 Diabetes KKAy Mice Treated with Couplet Medicines of Scutellariae Radix and Coptidis Rhizoma Based on nano-LC-LTQ/Orbitrap-MS[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(17): 126-131.
LIU Tong-tong, XIAO Qin, SHENG Jun-qing, et al. Screening of Differentially Expressed Proteins in Skeletal Muscle of Type 2 Diabetes KKAy Mice Treated with Couplet Medicines of Scutellariae Radix and Coptidis Rhizoma Based on nano-LC-LTQ/Orbitrap-MS[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(17): 126-131. DOI: 10.13422/j.cnki.syfjx.20181610.
Objective:To identify the differentially expressed proteins in skeletal muscle of type 2 diabetes KKAy mice from the model group and the administration group by nano-LC-LTQ/Orbitrap-MS
and speculate mechanism of couplet medicines of Scutellariae Radix and Coptidis Rhizoma against type 2 diabetes. Method:Twenty male KKAy mice aged 8-10 weeks were used. KKAy mice were given full-fledged high-fat diet and randomly divided into 2 groups
included Scutellariae Radix and Coptidis Rhizoma group(27 g·kg-1) and model group(the same amount of distilled water).Mice in the two groups were continuous intragastric administrated for 8 weeks.After the last administration for 1 hour
mice were sacrificed
skeletal muscle was taken and placed in -80℃ refrigerator.Skeletal muscles of the model group and administration group were ground in liquid nitrogen
lysate and protease inhibitors were added to extract the protein
the concentration of protein was determined
and then protein denaturation
reduction
alkylation
enzymolysis and desalting
the samples were identified by nano-LC-LTQ/Orbitrap-MS.Data were processed by Xcalibur 2.2 software and Proteome Discoverer 1.4.0.288
and Sieve v2.1(×64) was adopted to analyze and find relevant differential proteins with quantitative analysis of non-standard samples
differentially expressed proteins were screened and identified
and bioinformatics analysis of the differentially expressed proteins was carried out. Result:A total of 107 differentially expressed proteins were identified from Scutellariae Radix and Coptidis Rhizoma group and model group
and five differential proteins related to diabetes were screened out by biological analysis
including 4 up-regulated[5'-adenosine-activated protein kinase(AMPK)
adiponectin(APN)
etc] and 1 down-regulated[adipocyte type fatty acid binding protein4(FABP4)].The differential proteins mainly involved in inflammatory reactions
signal pathway regulation and other biological processes. Conclusion:Through effective comparison and analysis
we obtain some differentially expressed proteins involved in insulin signal transduction and other pathways
this paper can provide a new target for the study on the molecular mechanism of Scutellariae Radix and Coptidis Rhizoma against insulin resistance.