Correlation Between Autophagy-related Proteins in Peripheral Blood Mononuclear Cells and Inflammatory Markers in Hemodialysis Patients
Trial Snapshot
- Phase
- Not Applicable
- Status
- Completed
- Sponsor
- Enrollment
- 62
- Locations
- 1
- Primary Endpoint
- Levels of the autophagy-associated proteins microtubule-associated proteins 1A/1B light chain 3A (LC3I/II)
Study Overview
Brief Summary
One recent study demonstrated impaired autophagy in patients receiving hemodialysis (HD). To clarify whether this alteration is related to the inflammatory state in HD patients, we focused on basal autophagy in peripheral blood mononuclear cells (PBMCs) of HD patients with and without inflammation and controls. PBMCs were harvested using Ficoll density gradient centrifugation . Levels of the autophagy-associated proteins ubiquitin-binding protein p62 (p62), microtubule-associated proteins 1A/1B light chain 3A (LC3I/II) and beclin-1 in PBMCs will be detected by western blotting. Enzyme-linked immunosorbent assay kits will be used to detect the serum concentrations of interleukin (IL)-6, liposaccharide-binding protein (LBP) and tumor necrosis factor (TNF)-α.
Detailed Description
Study Design and Population
Study Design We shall enroll 30 healthy volunteers as the control group. 180 sex- and age-matched hemodialysis patients will be enrolled in this study. To be included in the study, patients have to be at least 20-years-old and on outpatient hemodialysis (HD) for at least 3 months. Patients are excluded if they had malignancy, severe liver disease (bilirubin >1.6 mg/dl), uncontrolled hypertension, severe obesity (BMI >35), currently on carbamazepine, statins or immunosuppressive agents. The medical record is thoroughly reviewed for each subject by a collaborating physician in the study. All subjects will provide written informed consent to participate and the protocol will be send to the institutional review boards of Tungs' Taichung Metroharbour Hospital
Study Parameters Predialysis blood samples are obtained on a mid-week day. Within 30 min after sampling, the remaining blood is centrifuged at 3,000 g for 10 min, immediately aliquoted and frozen at -80°C until further analysis.
Cytokine assays The following markers of inflammation and hemostasis are determined by ELISA in patients' sera: interleukin-6 (Quantikine, R&D Systems,) and plasma concentrations of CRP in a highly sensitive assay (hsCRP).
Analysis of lipids/lipoproteins Total cholesterol, highdensity lipoprotein (HDL) cholesterol, and triglyceride concentrations are determined enzymatically. Plasma glucose is measured by a glucose-oxidase method.
Study Design
- Study Type
- Observational
- Observational Model
- Case Control
- Time Perspective
- Cross Sectional
Eligibility Criteria
- Ages
- 20 Years to 90 Years (Adult, Older Adult)
- Sex
- All
- Accepts Healthy Volunteers
- Yes
Inclusion Criteria
- •Both sexes aged between 20-90 years.
- •Received stable hemodialysis at least 3 months.
- •Written informed consent.
Exclusion Criteria
- •patients with severe infections, severe heart disease and liver disease, malignancy, autoimmune disorders, severe malnutrition, or clinical conditions requiring oral nutrition supplements;
- •Inability to follow protocol.
- •Pregnancy or wishing/trying to get pregnant
Outcomes
Primary Outcomes
Levels of the autophagy-associated proteins microtubule-associated proteins 1A/1B light chain 3A (LC3I/II)
Time Frame: 1 years
microtubule-associated proteins 1A/1B light in PBMCs will be detected by western blotting.
Levels of the autophagy-associated proteins beclin-1
Time Frame: 1 years
beclin-1 in PBMCs will be detected by western blotting.
Levels of the autophagy-associated proteins ubiquitin-binding protein p62 (p62)
Time Frame: 1 years
ubiquitin-binding protein p62 (p62) in PBMCs will be detected by western blotting.
Secondary Outcomes
- Measurements of TNF-α(1 years)
- Analysis of biomarkers of IL-6(1 years)
- Analysis of biomarkers of Liposaccharide-binding protein (LBP)(1 years)