{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST001504","ANALYSIS_ID":"AN002492","VERSION":"1","CREATED_ON":"October 9, 2020, 12:20 pm"},

"PROJECT":{"PROJECT_TITLE":"Metabolomics reveals the protective effect of isosteviol sodium against multiple organ injury in septic mice","PROJECT_SUMMARY":"Sepsis is a severe inflammatory disorder that can lead to multiple organ injury. Isosteviol sodium (STV-Na) is a terpenoid derived from stevioside that exerts anti-inflammatory, antioxidant and anticancer activities. However, the influence of STV-Na on sepsis remains unknown. Here, we assessed the potential effects of STV-Na on sepsis and multiple organ injury induced by lipopolysaccharide (LPS). We found that STV-Na increased the survival rate of mice treat with LPS, significantly improved the functions of the heart, lung, liver, and kidney, and reduced the production of inflammatory cytokines. Moreover, Multiorgan metabolomics analysis demonstrated that glutathione metabolism, purine metabolism, glycerophospholipid metabolism and pantothenate and CoA biosynthesis, were significantly altered by STV-Na. This study provides novel insights into the metabolite changes of multiple organ injury in septic mice, which may help characterize the underlying mechanism and provide an improved understanding of the therapeutic effects of STV-Na on sepsis.","INSTITUTE":"Guangdong University of Technology","LAST_NAME":"Wang","FIRST_NAME":"Shanping","ADDRESS":"No. 100, Waihuan Xilu, Guangzhou Higher Education Mega Center, Panyu District,","EMAIL":"shanpingwang@outlook.com","PHONE":"15521002792"},

"STUDY":{"STUDY_TITLE":"Metabolomics reveals the protective effect of isosteviol sodium against multiple organ injury in septic mice - Spleen","STUDY_SUMMARY":"Sepsis is a severe inflammatory disorder that can lead to multiple organ injury. Isosteviol sodium (STV-Na) is a terpenoid derived from stevioside that exerts anti-inflammatory, antioxidant and anticancer activities. However, the influence of STV-Na on sepsis remains unknown. Here, we assessed the potential effects of STV-Na on sepsis and multiple organ injury induced by lipopolysaccharide (LPS). We found that STV-Na increased the survival rate of mice treat with LPS, significantly improved the functions of the heart, lung, liver, and kidney, and reduced the production of inflammatory cytokines. Moreover, Multiorgan metabolomics analysis demonstrated that glutathione metabolism, purine metabolism, glycerophospholipid metabolism and pantothenate and CoA biosynthesis, were significantly altered by STV-Na. This study provides novel insights into the metabolite changes of multiple organ injury in septic mice, which may help characterize the underlying mechanism and provide an improved understanding of the therapeutic effects of STV-Na on sepsis.","INSTITUTE":"Guangdong University of Technology","LAST_NAME":"Wang","FIRST_NAME":"Shanping","ADDRESS":"No. 100, Waihuan Xilu, Guangzhou Higher Education Mega Center, Panyu District,","EMAIL":"shanpingwang@outlook.com","PHONE":"15521002792"},

"SUBJECT":{"SUBJECT_TYPE":"Mammal","SUBJECT_SPECIES":"Mus musculus","TAXONOMY_ID":"10090"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"wild-type",
"Sample ID":"Ctrl-1",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"Ctrl-W-1"}
},
{
"Subject ID":"wild-type",
"Sample ID":"Ctrl-2",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"Ctrl-W-2"}
},
{
"Subject ID":"wild-type",
"Sample ID":"Ctrl-3",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"Ctrl-W-3"}
},
{
"Subject ID":"wild-type",
"Sample ID":"Ctrl-4",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"Ctrl-W-4"}
},
{
"Subject ID":"wild-type",
"Sample ID":"Ctrl-5",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"Ctrl-W-5"}
},
{
"Subject ID":"wild-type",
"Sample ID":"Ctrl-6",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"Ctrl-W-6"}
},
{
"Subject ID":"wild-type",
"Sample ID":"Ctrl-7",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"Ctrl-W-7"}
},
{
"Subject ID":"wild-type",
"Sample ID":"Ctrl-8",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"Ctrl-W-8"}
},
{
"Subject ID":"wild-type",
"Sample ID":"Ctrl-9",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"Ctrl-W-9"}
},
{
"Subject ID":"wild-type",
"Sample ID":"LPS-1",
"Factors":{"Treatment":"Model"},
"Additional sample data":{"RAW_FILE_NAME":"LPS-W-1"}
},
{
"Subject ID":"wild-type",
"Sample ID":"LPS-2",
"Factors":{"Treatment":"Model"},
"Additional sample data":{"RAW_FILE_NAME":"LPS-W-2"}
},
{
"Subject ID":"wild-type",
"Sample ID":"LPS-3",
"Factors":{"Treatment":"Model"},
"Additional sample data":{"RAW_FILE_NAME":"LPS-W-3"}
},
{
"Subject ID":"wild-type",
"Sample ID":"LPS-4",
"Factors":{"Treatment":"Model"},
"Additional sample data":{"RAW_FILE_NAME":"LPS-W-4"}
},
{
"Subject ID":"wild-type",
"Sample ID":"LPS-5",
"Factors":{"Treatment":"Model"},
"Additional sample data":{"RAW_FILE_NAME":"LPS-W-5"}
},
{
"Subject ID":"wild-type",
"Sample ID":"LPS-6",
"Factors":{"Treatment":"Model"},
"Additional sample data":{"RAW_FILE_NAME":"LPS-W-6"}
},
{
"Subject ID":"wild-type",
"Sample ID":"LPS-7",
"Factors":{"Treatment":"Model"},
"Additional sample data":{"RAW_FILE_NAME":"LPS-W-7"}
},
{
"Subject ID":"wild-type",
"Sample ID":"LPS-8",
"Factors":{"Treatment":"Model"},
"Additional sample data":{"RAW_FILE_NAME":"LPS-W-8"}
},
{
"Subject ID":"wild-type",
"Sample ID":"LPS-9",
"Factors":{"Treatment":"Model"},
"Additional sample data":{"RAW_FILE_NAME":"LPS-W-9"}
},
{
"Subject ID":"wild-type",
"Sample ID":"STV-1",
"Factors":{"Treatment":"Treatment"},
"Additional sample data":{"RAW_FILE_NAME":"STV-W-1"}
},
{
"Subject ID":"wild-type",
"Sample ID":"STV-2",
"Factors":{"Treatment":"Treatment"},
"Additional sample data":{"RAW_FILE_NAME":"STV-W-2"}
},
{
"Subject ID":"wild-type",
"Sample ID":"STV-3",
"Factors":{"Treatment":"Treatment"},
"Additional sample data":{"RAW_FILE_NAME":"STV-W-3"}
},
{
"Subject ID":"wild-type",
"Sample ID":"STV-4",
"Factors":{"Treatment":"Treatment"},
"Additional sample data":{"RAW_FILE_NAME":"STV-W-4"}
},
{
"Subject ID":"wild-type",
"Sample ID":"STV-5",
"Factors":{"Treatment":"Treatment"},
"Additional sample data":{"RAW_FILE_NAME":"STV-W-5"}
},
{
"Subject ID":"wild-type",
"Sample ID":"STV-6",
"Factors":{"Treatment":"Treatment"},
"Additional sample data":{"RAW_FILE_NAME":"STV-W-6"}
},
{
"Subject ID":"wild-type",
"Sample ID":"STV-7",
"Factors":{"Treatment":"Treatment"},
"Additional sample data":{"RAW_FILE_NAME":"STV-W-7"}
},
{
"Subject ID":"wild-type",
"Sample ID":"STV-8",
"Factors":{"Treatment":"Treatment"},
"Additional sample data":{"RAW_FILE_NAME":"STV-W-8"}
},
{
"Subject ID":"wild-type",
"Sample ID":"Dex-1",
"Factors":{"Treatment":"Positive"},
"Additional sample data":{"RAW_FILE_NAME":"Dex-W-1"}
},
{
"Subject ID":"wild-type",
"Sample ID":"Dex-2",
"Factors":{"Treatment":"Positive"},
"Additional sample data":{"RAW_FILE_NAME":"Dex-W-2"}
},
{
"Subject ID":"wild-type",
"Sample ID":"Dex-3",
"Factors":{"Treatment":"Positive"},
"Additional sample data":{"RAW_FILE_NAME":"Dex-W-3"}
},
{
"Subject ID":"wild-type",
"Sample ID":"Dex-4",
"Factors":{"Treatment":"Positive"},
"Additional sample data":{"RAW_FILE_NAME":"Dex-W-4"}
},
{
"Subject ID":"wild-type",
"Sample ID":"Dex-5",
"Factors":{"Treatment":"Positive"},
"Additional sample data":{"RAW_FILE_NAME":"Dex-W-5"}
},
{
"Subject ID":"wild-type",
"Sample ID":"Dex-6",
"Factors":{"Treatment":"Positive"},
"Additional sample data":{"RAW_FILE_NAME":"Dex-W-6"}
},
{
"Subject ID":"wild-type",
"Sample ID":"Dex-7",
"Factors":{"Treatment":"Positive"},
"Additional sample data":{"RAW_FILE_NAME":"Dex-W-7"}
},
{
"Subject ID":"wild-type",
"Sample ID":"Dex-8",
"Factors":{"Treatment":"Positive"},
"Additional sample data":{"RAW_FILE_NAME":"Dex-W-8"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Samples of blood were gathered in heparinized tubes and then spun down at 3000 rpm at 4 °C for 10 min to obtain supernatant of plasma samples for subsequent preparation. Heart, lung, liver, spleen and kidney samples were homogenized in a five-fold volume of normal saline, respectively, and spun down at 12,000 rpm at 4 °C for 20 min to acquire supernatants for further preparation.","COLLECTION_PROTOCOL_FILENAME":"shanpingwang_Collection_Protocol.docx","SAMPLE_TYPE":"Spleen"},

"TREATMENT":{"TREATMENT_SUMMARY":"After being acclimatized for 1 week, the mice were separated, at random, into four groups: 1) saline + saline; 2) saline + LPS; 3) STV-Na+ LPS; and 4) dexamethasone (Dex) + LPS. Mice were intraperitoneally administered saline (0.1 mL/10 g) or STV-Na (5, 10, 20 mg/kg) and Dex (10 mg/kg) two times per day every 12 h for 3 consecutive days, and one hour after the first intraperitoneal injection on day 3, saline (0.1 mL/g body weight) or LPS from E. coli (0111: B4, 20 mg/kg) was intraperitoneally administered.","TREATMENT_PROTOCOL_FILENAME":"shanpingwang_Treatment_Protocol.docx"},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"A total of 160 µL of MTBE solution (methyl-T-butyl-ether: methanol: water, 6/3/1, v/v/v) was applied to 40 µL of the plasma or tissue homogenate supernatant, vortexed for 30 min at 4°C and spun at 12,000 rpm for 30 min. Two extract components were produced: an organic hydrophobic layer and a hydrophilic layer. These two extracts were vacuum-dried and dissolved in 0.1% (v/v) formic acid in water (45 µL), followed by analysis. The pooled quality control (QC) samples including whole plasma and tissues were utilized for monitoring data acquisition performance throughout the analysis. Finally, 6 duplicate QC samples were prepared and injected at the start of the sequence, and after each of the six tissue samples was inserted, the QC samples were added to determine system stability.","SAMPLEPREP_PROTOCOL_FILENAME":"shanpingwang_Sampleprep_protocol.docx"},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_SUMMARY":"Chromatographic separations were conducted utilizing a Waters BEH C18, 2.1 mm×50 mm 1.7 µm particle column with a Dionex Ultimate 3000 UHPLC system from Thermo Fisher Scientific (CA, USA). The mobile phase encompassed water with 0.1% v/v formic acid (A) and acetonitrile with 0.1% v/v formic acid (B). Columns were kept at 40°C and eluted using a linear gradient: 2-30% B at 0-4 min, 30-40% B at 4-5 min, 40% B at 5-8 min, 40-60% B at 8-10 min, 60-100% B at 10-17 min, 100% B at 17-19 min, 100-2% B at 19-19.1 min, and 2% B at 19.1-25 min. To increase the amount of metabolites and save experimental time, a new sampling method was used to detect both the organic phase and the aqueous phase extracts (Qiuhui Xuan et al., 2018; Shanping Wang et al., 2019): 5 µL of organic phase extracts were first loaded without running the elution gradient, which lasted for one minute at the initial mobile phase, and then 5 µL of the aqueous phase extracts were added to the same column in order to start running the elution gradient using a 0.4 mL/min flow rate.","CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Thermo Dionex Ultimate 3000","COLUMN_NAME":"Waters Acquity BEH C18 (50 x 2.1mm, 1.7 um)","METHODS_FILENAME":"shanpingwang_Chromatography_Methods.docx"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS","ANALYSIS_PROTOCOL_FILE":"shanpingwang_Analysis_Protocol.docx"},

"MS":{"INSTRUMENT_NAME":"Bruker TIMS TOF","INSTRUMENT_TYPE":"QTOF","MS_TYPE":"ESI","ION_MODE":"POSITIVE","MS_COMMENTS":"MSMS Progenesis QI 2.1 software EZinfo 3.0 software","MS_RESULTS_FILE":"ST001504_AN002492_Results.txt UNITS:Intensity Has m/z:No Has RT:Yes RT units:Seconds"}

}