Nghiên cứu nồng độ SERP5, RBP4, IL 18 huyết thanh ở bệnh nhân đái tháo đường týp 2 TT TIENG ANH

MINISTRY OF DEPARTMENT OF DEFENSE EDUCATION AND TRAINING MILITARY MEDICAL UNIVERSITY NGUYEN VIET DUNG RESEARCH THE LEVEL SFRP5, RBP4, IL-18 SERUM IN TYPE II DIABETES PATIENTS Specialization: Internal Medicine Code: 9.72.01.07 SUMMARY THESIS OF PHD MEDICINE HA NOI – 2021 THESIS COMPLETING AT MILITARY ACADEMY PhD Assoc Prof DOAN VAN DE Prof Dr NGUYEN LINH TOAN SCIENTIFIC GUIDE: Reviewer 1: PhD Assoc Reviewer 2: PhD Prof VU THI THANH HUYEN NGUYEN VAN DO Reviewer 3: PhD Assoc Prof NGUYEN KIM LUONG The thesis will be defended in front of The Thesis Judging Committee at… The thesis can be found at: National Library Military Medical Academy Library INTRODUCTION Diabetes type is one of the most common chronic diseases in the world, which is rapidly increasing and becoming a great concern issue of public health, especially for developing countries According to a survey by the International Diabetes Federation (IDF), in 2017 there were about 425 million people (age 20-79) with diabetes accounting for 8.8% and 451 million people if the age range is expanded from 18 -99 Patients type diabetes are often overweight, obese and having a gain in adipose tissue, concentrated in the abdomen and visceral area Recent studies have shown that adipose tissue not only plays a vital in energy storage, but also has an important endocrine function Adipose tissue secretes a lot of hormones and chemical mediators with important roles in the regulation of energy metabolism, retinol binding protein (RBP4), MCP-1, has been shown to be related with the mechanism of inducing the metabolic disorders body, some are risk factors and some are agents against type diabetes, metabolic syndrome and cardiovascular disease Among them, SFRP5 is known to be an anti-inflammatory adipokine and RBP4, IL18 is a pro-inflammatory adipokines that are strongly associated with lowgrade chronic inflammation, overweight, obesity, type diabetes, atherosclerosis arteries, … However, the abnormal increase and decrease in levels of SFRP5, RBP4, IL-18 and its role in the pathogenesis of overweight, obesity, and type diabetes are controversial and need to be studied and clarified In Vietnam, there have been some authors studying about adipose tissues: adiponectin, leptin, MCP-1, TNF-α, IL-6, IL-10, hs-CRP, in overweight, obesity, pre-diabetes, type diabetes, hypertension However, no studies addressing the role and the relationship of SFRP5, RBP4, IL-18 in type diabetes patients Therefore, studying the changes in concentrations and their relationship in patients with type diabetes is a necessary and meaningful study science and practice Stemming from the above fact, we conducted a study on the topic "Studying serum levels of SFRP5, RBP4, IL-18 in patients with type diabetes" with objectives: Investigation of serum SFRP5, RBP4, IL-18 levels in type diabetes patients Analysis of the relationship between level serum SFRP5, RBP4, IL-18 with some clinical and subclinical characteristics and insulin resistance in type diabetes patients Practical significance and new contributions - The thesis confirms that the thesis is one of the research works at the doctoral level that has contributed to the specialty of Internal Medicine Endocrinology This is a new study that has never been done in the country - Research results have described serum levels of SFRP5, RBP4, IL-18 in patients with type diabetes Analysis of the relationship between serum SFRP5, RBP4, IL-18 levels with some clinical characteristics Clinical, subclinical and insulin resistance status in patients with type diabetes - Through quantification of these adipokin levels, we give specific values of SFRP5, RBP4, IL-18 concentrations in people with and without type diabetes in Vietnam At the same time, based on the correlation between these adipokin levels and some cardiovascular-metabolic risk factors, it is possible to identify their effects on the body in patients with type diabetes Thesis structure The thesis is presented in 133 pages, including: pages of problem statement, 34 pages of overview, 19 pages of research objects and methods, 30 pages of research results, 45 pages of discussion, pages of conclusion and 01 page of recommendations The thesis has 14 images, 42 tables, 15 charts, 01 diagram, there are 177 references including 22 Vietnamese documents, 155 English documents CHAPTER 1: OVERVIEW 1.1 OVERVIEW OF SFRP5, RBP4, IL-18 1.1.1 Secreted Fizzled - Related Protein (SFRP5) In recent years, a number of studies on SFRP5 have been conducted in the world, both experimentally and clinically, and have proven that SFRP5 has anti-inflammatory effects and decreased SFRP5 levels are the cause of metabolic syndrome (HCCH), obesity and cardiovascular disease SFRP5 has a role in regulating glucose metabolism, potentially reducing the risk of HCC, type diabetes, cardiovascular disease, etc In addition, SFRP5 also works to reduce belly fat and promote lipoprotein oxidation SFRP5 in insulin resistance and type diabetes: Study at the cellular level, SFRP5 is upregulated by pancreatic islets of obese rodents and humans Administration of SFRP5 reduced proliferation in pancreatic islet cells and experimental studies showed that overexpression of SFRP5 resulted in higher serum fasting insulin levels in both lean and lean mice obese mice 1.1.2 Retinol binding protein (RBP4) RBP4, a retinol transporter, increases insulin resistance and contributes to an increased risk of diabetes To determine the mechanism underlying RBP4induced insulin resistance, we found that an increase in RBP4 induces adipose tissue inflammation by activating innate immunity that induces an adaptive immune response Mice with excessive levels of RBP4 (RBP4-Ox) exhibit insulin resistance and glucose intolerance, and increased CD4 T-cell and macrophage infiltration in inflammatory adipose tissue 1.1.3 Interlekin 18 (IL-18) Role of IL-18 in Type Diabetes: In an experimental study in NOD (nonobese diabetic) mice, the authors reported that IL-18 was implicated in the development of diabetes in mice NOD Indeed, the IL-18 gene is located near Idd2, which is one of the genes involved in the development of diabetes in NOD mice In addition, in parallel with IL-12 mRNA expression, IL-18 mRNA expression was increased in the pancreas of NOD mice after inducing diabetes with cyclophosphamide The author also showed that exogenous IL-18 can modulate autoimmunity against pancreatic β-cells through Th1-directed immune response and that exogenous IL-18 not only accelerates atherosclerosis in mice but also accelerated the features of diabetes/ Author Everett B.M and CS demonstrated that hyperglycemia directly promotes increased IL-18 in humans through an oxidative mechanism 1.2 STUDYING ON SFRP5, RBP4, IL-18 IN TYPE DIABETES PATIENTS 1.2.1.Research situation on SFRP5, RBP4, IL-18 in the world 1.2.1.1 Research status of SFRP5 in type diabetes patients Ouchi N et al reported that SFRP5-deficient mice had normal glucose tolerance when maintained on the diet, but there was a decline in insulin sensitivity when fed high-calorie diets Injection of recombinant SFRP5 to obese mice improves metabolic parameters such as insulin resistance and fatty liver disease Thus, the study of SFRP5 in adipose tissue is a potential target for the control of obesity-related abnormalities and glucose homeostasis 1.2.1.2 Research status of RBP4 in type diabetes patients Many clinical studies have demonstrated that RBP4 levels increase in proportion to body mass index (BMI) in both obese and non-diabetic patients It has also been shown to increase independently of body fat accumulation, which is more strongly associated with increased visceral fat than subcutaneous fat Significant weight loss with diet, exercise, or bariatric surgery have all been shown to reduce RBP4 levels and correlate with improvements in insulin sensitivity This depletion is more specific to leptin, adiponectin, IL6 or CRP transformation 1.2.1.3 Research status of IL-18 in type diabetes patients Studies have shown that blood levels of IL-18 are increased in patients with type diabetes and contribute to capillary disease such as nephropathy in type diabetes Furthermore, in a cohort study over time, increased IL-18 levels were shown to predict the development of type diabetes Also from the Dallas Heart Study, Andreas Zirlik and CS identified an association between IL-18 levels and insulin resistance methods, such as HOMA-IR, and hyperglycemia IL-18 levels were implicated in the determination of fasting glucose levels, however this association occurred only when adjusted for HOMA-IR Furthermore, for subjects with diabetes, the correlation between IL-18 and obesity was not significant, suggesting that insulin resistance is a determinant of IL-18 1.2.2 Research status on adipose tissue cytokines and SFRP5, RBP4, IL18 in Vietnam Currently, in Vietnam, there are no studies on the change of concentration and relationship of SFRP5, RBP4, IL-18 with clinical and subclinical characteristics in type diabetes patients Therefore, the study Studying the change in concentration and relationship of these adipokines in type diabetes patients is a necessary study, having scientific and practical significance Through quantification of these adipokin levels, we give specific values of SFRP5, RBP4, IL-18 concentrations in people with and without type diabetes in Vietnam At the same time, based on the correlation between these adipokin levels and some cardiovascularmetabolic risk factors, it is possible to identify their effects on the body in type diabetes patients CHAPTER 2: RESEARCH SUBJECTS AND METHODS 2.1 RESEARCH SUBJECTS The study included 323 study participants divided into groups Convenience, randomization, purposeful sampling of 184 patients with type diabetes (according to ADA 2012 criteria) and 139 people without diabetes as a control group for outpatient examination and treatment at Nghe An Endocrinology Hospital, Protecting and Taking care of the health of Thai Nguyen officials Exclusion criteria - Group of patients with type diabetes: Having acute infection; Diabetes with severe or multiple complications; Containing acute cerebral stroke, exacerbation of hypertension; Patients with cirrhosis, liver failure, heart failure, severe renal failure, chronic obstructive pulmonary disease, gout or other metabolic diseases - Control group: Having acute infection; Patients with cirrhosis, liver failure, heart failure, severe renal failure, chronic obstructive pulmonary disease, gout or other metabolic diseases; Having other co-morbidities such as: severe infections, hepatitis, tuberculosis, autoimmune diseases or diseases affecting glucose metabolism such as acromegaly, hyperthyroidism syndrome, Cushing's, nephrotic syndrome , Polycystic Ovary, … ; Are taking some drugs such as: glucocorticoid group, non-steroid, thiazide diuretics, oral contraceptives in the past month 2.2 METHODS 2.2.1 Research design * Research method: Prospective, descriptive, cross-sectional, with comparison with control group * Research location: The study was carried out at Nghe An Endocrinology Hospital, Thai Nguyen Provincial Department of Health Protection and Care, SFRP5, RBP4, IL-18 serum analyzed at the Department of Physiology, Military Medical Academy * Research period: Collecting data from October 2013 to June 2018 2.2.2 Quantification of SFRP5, RBP4 and IL-18 a) Principle: Implemented by Sandwich-type enzyme-linked immunosorbent assay (ELISA) method based on specific reaction between antibody attached at the bottom of the well with cytokine antigen (SFRP5, RBP4 and IL-18) present in patient serum, ELISA kit SFRP5, RBP4, IL-18 of Cloud Clone Corp, USA, reading concentration on ELISA machine of Diagnostic Automation, USA b) Steps: - Blood collection: Take venous blood on fasting in the morning, the amount of 5ml is put into a tube containing anticoagulant, centrifuge at 3000rpm to separate the serum, then store it in a cold freezer right away (temperature 80 0C) Serum samples, after being fully collected at each facility, are transferred to the Department of Pathophysiology - Military Medical Academy (test tubes are stored in a specialized cooler to preserve vaccines, medical biological products, and medical specimens) stored between layers RESEARCH SUBJECTS containing dry ice, transport time (n=323) from 1.5 hours to hours), then continue to be stored at sonic depth - 80 0C until testing - Place: Department of Pathophysiology, Military Medical Academy - Perform quantitative steps according to the manufacturer's instructions on the ELISA machine of Diagnostic Automation, USA TYPE DIABETES RESEARCH DIAGRAM CONTROL GROUP (n=139) GROUP (n=184) QUESTIONS, CLINICAL EXAMINATIN, TEST DIAGNOSTIC TYPE DIABETES - TCBP, KTCBP CERTIFICATE - TCBP, KTCBP QUANTIFICATION SFPP5( n=279), RBP4(n=311), IL-18(n=323) COMPARISON Analysis of the relationship between SFRP5, RBP4, IL-18 with some clinical and subclinical characteristics in type diabetes patients Investigation of serum SFRP5,RBP4,IL-18 level in type diabetes CONCLUSION CONCLUSION PETITION CHAPTER 3: RESULTS 3.1 GENERAL CHARACTERISTICS OF STUDY SUBJECTS The proportion of study subjects by age groups in the group with type diabetes and the control group did not differ with p>0.05 The prevalence of diabetes increases with age, mainly ≥ 50 years old However, the mean age of the group with type diabetes and the control group was different with p