Metabolism of Branched-chain Amino Acids in Monocyte/Macrophage During Sepsis

Study Overview
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Brief Summary
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The current project was designed to examine the metabolic level of branched-chain amino acids in plasm and monocyte/macrophage, and its role in immune dysfunction during sepsis.

Detailed Description
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In this study, the investigators will collect the peripheral blood of 1,3,7,14 day after diagnose of sepsis. Then, the concentration of branched-chain amino acids(BCAAs) in monocytes and plasma, and the expression of BCAAs metabolic enzymes in monocytes will be determined. Additionally, the inflammatory cytokines, e.g., Interleukin-1β (IL-1β), Tumor necrosis factor-α (TNF-α), Interleukin-10 (IL-10）, the chemokines macrophage inflammatory proteins 1α and 1β (MIP-1α and 1β) in the peripheral blood and the phagocytic function of monocytes will also be detected. The current project was designed to examine the metabolic level of branched-chain amino acids in plasm and monocyte/macrophage, and its role in immune dysfunction during sepsis.

Official Title
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Metabolism of Branched-chain Amino Acids in Monocyte and Macrophage During Sepsis

Conditions
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Sepsis Syndrome

Intervention / Treatment
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* Other: Metabolic level of BCAAs in monocytes and macrophages


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: patients diagnosed with sepsis, with or without type 2 diabetes healthy volunteers Exclusion Criteria: patients with immunodeficient diseases patients who accept glucocorticoid or immunosuppressant patients with an age younger than 18 years patients who cannot provide the informed consent

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 70 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Sepsis with type 2 diabetes<br>Sepsis patients with type 2 diabetes | Other: Metabolic level of BCAAs in monocytes and macrophages<br>* Metabolic level of BCAAs in monocytes and macrophages<br>|
| Sepsis without type 2 diabetes<br>Sepsis patients without type 2 diabetes | Other: Metabolic level of BCAAs in monocytes and macrophages<br>* Metabolic level of BCAAs in monocytes and macrophages<br>|
| Volunteers<br>Healthy volunteers | Other: Metabolic level of BCAAs in monocytes and macrophages<br>* Metabolic level of BCAAs in monocytes and macrophages<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Intracellular levels of branched-chain amino acids in monocytes | BCAAs Assay Kit will be used to measure the intracellular levels of BCAAs in monocytes. | up to 14 day after diagnose of sepsis | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Plasma levels of branched-chain amino acids | BCAAs Assay Kit will be used to measure the plasma levels of BCAAs. | 1,3,5,7,10,14 day after diagnose of sepsis | 
| The expression of branched-chain amino acid aminotransferase in monocytes | The expression of branched-chain amino acid aminotransferase in monocytes will be determined by qPCR and western blot. | 1,3,5,7,10,14 day after diagnose of sepsis | 
| The expression of branched-chain α-ketoacid dehydrogenase in monocytes | The expression of branched-chain α-ketoacid dehydrogenase in monocytes will be determined by qPCR and western blot. | 1,3,5,7,10,14 day after diagnose of sepsis | 
| The expression of PP2Cm in monocytes | The expression of mitochondrial-targeted phosphatase, PP2Cm in monocytes will be determined by qPCR and western blot. | 1,3,5,7,10,14 day after diagnose of sepsis | 
| The plasma level of IL-1β | The plasma level of IL-1β will be determined by ELISA kit. | 1,3,5,7,10,14 day after diagnose of sepsis. | 
| The plasma level of TNF-α | The plasma level of TNF-α will be determined by ELISA kit. | 1,3,5,7,10,14 day after diagnose of sepsis. | 
| The plasma level of IL-10 | The level of IL-10 will be determined by ELISA kit. | 1,3,5,7,10,14 day after diagnose of sepsis. | 
| The plasma level of chemokine MIP-1α. | The plasma level of chemokine MIP-1α will be determined by ELISA kit. | 1,3,5,7,10,14 day after diagnose of sepsis. | 
| The plasma level of chemokine MIP-1β. | The plasma level of chemokine MIP-1β will be determined by ELISA kit. | 1,3,5,7,10,14 day after diagnose of sepsis. | 
| The phagocytic index of monocytes. | The phagocytic index of monocytes will be determined by a E.coli Phagocytosis Assay Kit. | 1,3,5,7,10,14 day after diagnose of sepsis. |