#METABOLOMICS WORKBENCH Lxy2024_20250207_002224 DATATRACK_ID:5598 STUDY_ID:ST003733 ANALYSIS_ID:AN006125 PROJECT_ID:PR002319 VERSION 1 CREATED_ON February 16, 2025, 7:56 pm #PROJECT PR:PROJECT_TITLE Proteomics and metabolomics analysis of Mongolian acupuncture therapy for type 2 PR:PROJECT_TITLE diabetes mellitus with atherosclerosis PR:PROJECT_TYPE Metabolomics analysis PR:PROJECT_SUMMARY The results of serum biochemical and H&E staining analysis suggest that PR:PROJECT_SUMMARY acupuncture therapy can improve cellular inflammatory factors, kidney function, PR:PROJECT_SUMMARY serum lipid function and upper carotid artery tissue lesions in rat of T2DM with PR:PROJECT_SUMMARY atherosclerosis, which is an effective treatment method. In addition, we applied PR:PROJECT_SUMMARY proteomics and metabolomics to comprehensively demonstrate that Mongolian PR:PROJECT_SUMMARY acupuncture therapy in T2DM with atherosclerosis mainly through regulation of PR:PROJECT_SUMMARY fatty acid metabolism. PR:INSTITUTE Inner Mongolia Medical University PR:DEPARTMENT School of Mongolian Medicine and Pharmacy PR:LAST_NAME Li PR:FIRST_NAME Xueyong PR:ADDRESS Hohhot 010110, P. R. China PR:EMAIL 1324612868@qq.com PR:PHONE 18947196075 #STUDY ST:STUDY_TITLE Proteomics and metabolomics analysis of Mongolian acupuncture therapy for type 2 ST:STUDY_TITLE diabetes mellitus with atherosclerosis ST:STUDY_SUMMARY The results of serum biochemical and H&E staining analysis suggest that ST:STUDY_SUMMARY acupuncture therapy can improve cellular inflammatory factors, kidney function, ST:STUDY_SUMMARY serum lipid function and upper carotid artery tissue lesions in rat of T2DM with ST:STUDY_SUMMARY atherosclerosis, which is an effective treatment method. In addition, we applied ST:STUDY_SUMMARY proteomics and metabolomics to comprehensively demonstrate that Mongolian ST:STUDY_SUMMARY acupuncture therapy in T2DM with atherosclerosis mainly through regulation of ST:STUDY_SUMMARY fatty acid metabolism. ST:INSTITUTE Inner Mongolia Medical University ST:LAST_NAME Li ST:FIRST_NAME Xueyong ST:ADDRESS Hohhot 010110, P. R. China ST:EMAIL 1324612868@qq.com ST:PHONE 18947196075 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Rattus norvegicus SU:TAXONOMY_ID 10116 SU:GENDER Male #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data SUBJECT_SAMPLE_FACTORS - Control1 Sample source:carotid artery tissue | Factor:Control RAW_FILE_NAME(Raw file name)=Pos_Control1.mzML Neg_Control1.mzML SUBJECT_SAMPLE_FACTORS - Control2 Sample source:carotid artery tissue | Factor:Control RAW_FILE_NAME(Raw file name)=Pos_Control2.mzML Neg_Control2.mzML SUBJECT_SAMPLE_FACTORS - Control3 Sample source:carotid artery tissue | Factor:Control RAW_FILE_NAME(Raw file name)=Pos_Control3.mzML Neg_Control3.mzML SUBJECT_SAMPLE_FACTORS - Control4 Sample source:carotid artery tissue | Factor:Control RAW_FILE_NAME(Raw file name)=Pos_Control4.mzML Neg_Control4.mzML SUBJECT_SAMPLE_FACTORS - Control5 Sample source:carotid artery tissue | Factor:Control RAW_FILE_NAME(Raw file name)=Pos_Control5.mzML Neg_Control5.mzML SUBJECT_SAMPLE_FACTORS - Mol1 Sample source:carotid artery tissue | Factor:Model RAW_FILE_NAME(Raw file name)=Pos_Mol1.mzML Neg_Mol1.mzML SUBJECT_SAMPLE_FACTORS - Mol2 Sample source:carotid artery tissue | Factor:Model RAW_FILE_NAME(Raw file name)=Pos_Mol2.mzML Neg_Mol2.mzML SUBJECT_SAMPLE_FACTORS - Mol3 Sample source:carotid artery tissue | Factor:Model RAW_FILE_NAME(Raw file name)=Pos_Mol3.mzML Neg_Mol3.mzML SUBJECT_SAMPLE_FACTORS - Mol4 Sample source:carotid artery tissue | Factor:Model RAW_FILE_NAME(Raw file name)=Pos_Mol4.mzML Neg_Mol4.mzML SUBJECT_SAMPLE_FACTORS - Mol5 Sample source:carotid artery tissue | Factor:Model RAW_FILE_NAME(Raw file name)=Pos_Mol5.mzML Neg_Mol5.mzML SUBJECT_SAMPLE_FACTORS - Acp1 Sample source:carotid artery tissue | Factor:Acupuncture RAW_FILE_NAME(Raw file name)=Pos_Acp1.mzML Neg_Acp1.mzML SUBJECT_SAMPLE_FACTORS - Acp2 Sample source:carotid artery tissue | Factor:Acupuncture RAW_FILE_NAME(Raw file name)=Pos_Acp2.mzML Neg_Acp2.mzML SUBJECT_SAMPLE_FACTORS - Acp3 Sample source:carotid artery tissue | Factor:Acupuncture RAW_FILE_NAME(Raw file name)=Pos_Acp3.mzML Neg_Acp3.mzML SUBJECT_SAMPLE_FACTORS - Acp4 Sample source:carotid artery tissue | Factor:Acupuncture RAW_FILE_NAME(Raw file name)=Pos_Acp4.mzML Neg_Acp4.mzML SUBJECT_SAMPLE_FACTORS - Acp5 Sample source:carotid artery tissue | Factor:Acupuncture RAW_FILE_NAME(Raw file name)=Pos_Acp5.mzML Neg_Acp5.mzML SUBJECT_SAMPLE_FACTORS - QC1 Sample source:carotid artery tissue | Factor:QC RAW_FILE_NAME(Raw file name)=Pos_QC1.mzML Neg_QC1.mzML SUBJECT_SAMPLE_FACTORS - QC2 Sample source:carotid artery tissue | Factor:QC RAW_FILE_NAME(Raw file name)=Pos_QC2.mzML Neg_QC2.mzML SUBJECT_SAMPLE_FACTORS - QC3 Sample source:carotid artery tissue | Factor:QC RAW_FILE_NAME(Raw file name)=Pos_QC2.mzML Neg_QC3.mzML #COLLECTION CO:COLLECTION_SUMMARY SD male rats (SPF, 130-150 g) were randomly selected as diabetic modals of DM CO:COLLECTION_SUMMARY (fed high-sugar and high-fat diet). After 6 weeks of induction of insulin CO:COLLECTION_SUMMARY resistance, STZ injection was diluted with sterile sodium citrate buffer (0.1 CO:COLLECTION_SUMMARY mol L-1, pH 4.2) and injected into mice at a subpathogenic dose (30 mg kg-1) CO:COLLECTION_SUMMARY tail vein to induce the occurrence of diabetes. After injection, caudal vein CO:COLLECTION_SUMMARY blood was collected on day 3, day 5 and day 7 respectively, and blood glucose CO:COLLECTION_SUMMARY exceeding 16.7 mmol L-1 was successful in the modeling of diabetic rats, and CO:COLLECTION_SUMMARY unsuccessful 15 rats were excluded. Then vitamin D3 injection was given to CO:COLLECTION_SUMMARY diabetic model rats with a total dose of 6×105 U kg-1 for 3 consecutive days, CO:COLLECTION_SUMMARY which damaged the integrity of the arterial wall of the rats, increased CO:COLLECTION_SUMMARY endothelial permeability, promoted the immersion and deposition of lipids and CO:COLLECTION_SUMMARY calcium in the vascular wall, caused the overload of arterial calcium, and CO:COLLECTION_SUMMARY promoted the degeneration and calcification of smooth muscle cells. Rats in the CO:COLLECTION_SUMMARY diabetic model group are continued to receive a high-fat diet for 4 weeks. At CO:COLLECTION_SUMMARY the end of the experiment, all rats were sacrificed, and the upper carotid CO:COLLECTION_SUMMARY artery tissue was sent for examination. CO:SAMPLE_TYPE carotid artery tissue #TREATMENT TR:TREATMENT_SUMMARY the upper carotid artery tissue of about 1 cm long was quickly taken, rinsed TR:TREATMENT_SUMMARY with normal saline, frozen with liquid nitrogen and stored at -80 ℃ until TR:TREATMENT_SUMMARY metabolite detection #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Samples were weighed before the extraction of metabolites and dried lyophilized SP:SAMPLEPREP_SUMMARY were ground in a 2 mL Eppendorf tube containing a 5 mm tungsten bead for 1 min SP:SAMPLEPREP_SUMMARY at 65 Hz in a Grinding Mill. Metabolites were extracted using 1 mL precooled SP:SAMPLEPREP_SUMMARY mixtures of methanol and water (v/v,4:1) and then placed for 1 h ultrasonic SP:SAMPLEPREP_SUMMARY shaking in ice baths. Subsequently, the mixture was placed at -20 °C for 1 h SP:SAMPLEPREP_SUMMARY and centrifuged at 14,000 g for 20 min at 4 °C. The supernatants were recovered SP:SAMPLEPREP_SUMMARY and concentrated to dryness in vacuum. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Shimadzu Nexera X2 CH:COLUMN_NAME Waters ACQUITY UPLC HSS T3 (100 x 2.1mm,1.8um) CH:SOLVENT_A 0.1% formic acid in water CH:SOLVENT_B 100% acetonitrile (ACN) CH:FLOW_GRADIENT The gradient was 0% buffer B for 2 min and was linearly increase to 48% in 4 CH:FLOW_GRADIENT min, and then up to 100% in4 min and maintained for 2 min, and then decreased to CH:FLOW_GRADIENT 0% buffer B in 0.1 min, with 3 min re-equilibration period employed. CH:FLOW_RATE 0.3 mL/min CH:COLUMN_TEMPERATURE 40 ℃ #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Q Exactive Plus Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS The electrospray ionization (ESI) with positive-mode and negative mode were MS:MS_COMMENTS applied for MS data acquisition separately. The instrument was set to acquire MS:MS_COMMENTS over the m/z range 70-1050 Da for full MS. The full MS scans were acquired at a MS:MS_COMMENTS resolution of 70,000 at m/z 200, and 17,500 at m/z 200 for MS/MS scan. The MS:MS_COMMENTS maximum injection time was set to for 100 ms for MS and 50 ms for MS/MS. The MS:MS_COMMENTS isolation window for MS2 was set to 2 m/z and the normalized collision energy MS:MS_COMMENTS (stepped) was set as 20, 30 and 40 for fragmentation. The raw MS data were MS:MS_COMMENTS processed using MS-DIAL for peak alignment, retention time correction and peak MS:MS_COMMENTS area extraction. MS:MS_RESULTS_FILE ST003733_AN006125_Results.txt UNITS:Peak area Has m/z:Yes Has RT:Yes RT units:Minutes #END