Correlation Between Autophagy-related Proteins in Peripheral Blood Mononuclear Cells and Inflammatory Markers in Hemodialysis Patients
试验速览
- 阶段
- 不适用
- 状态
- 已完成
- 发起方
- 入组人数
- 62
- 试验地点
- 1
- 主要终点
- Levels of the autophagy-associated proteins microtubule-associated proteins 1A/1B light chain 3A (LC3I/II)
研究概览
简要总结
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.
详细描述
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.
研究设计
- 研究类型
- Observational
- 观察模型
- Case Control
- 时间视角
- Cross Sectional
入排标准
- 年龄范围
- 20 Years 至 90 Years（Adult, Older Adult）
- 性别
- All
- 接受健康志愿者
- 是
入选标准
- •Both sexes aged between 20-90 years.
- •Received stable hemodialysis at least 3 months.
- •Written informed consent.
排除标准
- •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
结局指标
主要结局
Levels of the autophagy-associated proteins microtubule-associated proteins 1A/1B light chain 3A (LC3I/II)
时间窗: 1 years
microtubule-associated proteins 1A/1B light in PBMCs will be detected by western blotting.
Levels of the autophagy-associated proteins beclin-1
时间窗: 1 years
beclin-1 in PBMCs will be detected by western blotting.
Levels of the autophagy-associated proteins ubiquitin-binding protein p62 (p62)
时间窗: 1 years
ubiquitin-binding protein p62 (p62) in PBMCs will be detected by western blotting.
次要结局
- Measurements of TNF-α(1 years)
- Analysis of biomarkers of IL-6(1 years)
- Analysis of biomarkers of Liposaccharide-binding protein (LBP)(1 years)