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

• Conditions: Sepsis Syndrome
• Interventions: Other: Metabolic level of BCAAs in monocytes and macrophages

• Registration Number: NCT03842371
• Lead Sponsor: West China Hospital

Brief Summary

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

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.

Study Timeline

• Status Verified: 2019-02
• Study First Submitted: 2019-02-08
• Study Start: 2019-02-11
• Study First Submitted QC: 2019-02-14
• Last Update Submitted: 2019-02-14
• Study First Posted: 2019-02-15
• Last Update Posted: 2019-02-15
• Primary Completion: 2021-12-30
• Study Completion: 2022-02-28

Recruitment & Eligibility

• Status: UNKNOWN
• Sex: All
• Target Recruitment: 90

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

Study & Design

• Study Type: OBSERVATIONAL
• Study Design: Not specified

Arm && Interventions

Group	Intervention	Description
Sepsis without type 2 diabetes	Metabolic level of BCAAs in monocytes and macrophages	Sepsis patients without type 2 diabetes
Sepsis with type 2 diabetes	Metabolic level of BCAAs in monocytes and macrophages	Sepsis patients with type 2 diabetes
Volunteers	Metabolic level of BCAAs in monocytes and macrophages	Healthy volunteers

Primary Outcome Measures

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

Secondary Outcome Measures

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