Investigation of the Differential Expression and Diagnostic Value of Microrna-21-5p and Microrna-125b-5p in Patients with Type 2 Diabetes Mellitus and Mild Cognitive Impairment

Research Article

Austin Biomark Diagn. 2023; 5(1): 1027.

Investigation of the Differential Expression and Diagnostic Value of Microrna-21-5p and Microrna-125b-5p in Patients with Type 2 Diabetes Mellitus and Mild Cognitive Impairment

Yang Mingyue1,2*; Wang Jue1,2; Liu Enlin1,3; Zhang Shuxin1,2; Wang Gaoyuan5; Yang Shufang1,3,4

1Taizhou People’s Hospital Affiliated to Nanjing Medical University, China

2Dalian Medical University, Dalian, China

3Taizhou People’s Hospital, the Fifth Affiliated Hospital of Nantong University, China

4School of Medicine of Southeast University, China

5Graduate School of Nanjing University of Chinese Medicine, China

*Corresponding author: Yang Mingyue Taizhou People’s Hospital Affiliated to Nanjing Medical University, Taizhou, Jiangsu 225300, China. Email: yangsufang@njmu.edu.cn

Received: April 04, 2023 Accepted: May 03, 2023 Published: May 10, 2023

Abstract

Objective: To investigate the differential expression of microRNA-21-5p (miR-21-5p) and microRNA-125b-5p (miR-125b-5p) in patients with Type 2 Diabetes Mellitus (T2DM) and Mild Cognitive Impairment (MCI), and to evaluate their potential value as biomarkers in patients with T2DM and MCI (T2DM+MCI).

Methods: In this study, the Montreal Cognitive Assessment (MoCA) was used to assess cognitive function. The differentially expressed miRNAs were screened by second-generation high-throughput sequencing of 8 human whole blood samples. The 60 patients were verified by Real-time fluorescence quantitative Polymerase Chain Reaction (qRT-PCR).

Results: The expressions of miR-21-5p and miR-125b-5p in whole blood in T2DM+MCI group were significantly higher than those in T2DM+nMCI group. Spearman correlation analysis suggested that the expression levels of miR-21-5p and miR-125b-5p were negatively correlated with MOCA scores. ROC curve showed that the area under the curve of miR-21-5p was 0.724, the sensitivity was 0.633, the specificity was 0.833, the 95% confidence interval was 0.5955-0.854, and the optimal cut-off value was 7.457. The area under the curve of miR-125b-5p was 0.686, the sensitivity was 0.6, and the specificity was 0.767. The 95% confidence interval is 0.549-0.822, and the optimal cut-off value is 4.523.

Conclusions: miR-21-5p and miR-125b-5p were up-regulated in T2DM+MCI patients, and their expression levels were negatively correlated with MOCA scores. miR-21-5p and miR-125b-5p may be biomarkers for the diagnosis of T2DM+MCI patients.

Introduction

T2DM has been a major health hazard worldwide, and its global incidence continues to rise. Alzheimer's Disease (AD) is the most common neurodegenerative disease at present, accounting for nearly 80% of the global dementia population [1]. Dementia caused by Diabetes Mellitus (DM) can be divided into the following two types: One is neurocognitive dementia (VaD), which is directly related to DM. The other is that DM causes serious endocrine neurodegeneration -- AD. MCI is the stage that occurs between normal neurodegenerative aging and AD [2]. Studies have reported that DM patients are at risk of developing cognitive impairment, which is also known as “DM-related cognitive dysfunction” [3-5]. Severe cognitive impairment may be a new long-term complication of T2DM. Therefore, early identification of MCI can delay the onset of AD.

miRNAs are non-coding Rnas composed of 18-25 nucleotides [6], which have been reported as important targets for the diagnosis and treatment of AD [7,8]. Studies have shown that highly expressed or specifically expressed miRNAs found in brain tissues account for more than half of detectable miRNAs [9]. Studies have shown that cognitive impairment is related to hippocampus of rats [10]. Hippocampus tissue is the golden standard for cognitive impairment, but it is difficult to achieve clinically. Cerebrospinal fluid detection requires lumbar puncture, which is an invasive operation and cannot be obtained from a large number of patients. Since detecting blood samples is a relatively economical and minimally invasive method, the measurement of miRNAs expression patterns in blood has become an effective new diagnostic tool and a promising prognostic biomarker for the diagnosis of neurological diseases. The purpose of this study was to investigate the differential expression of miRNAs in T2DM+MCI patients and to evaluate their potential value as biomarkers.

Materials and Methods

Research Objects

This research plan was reviewed and approved by the Ethics Committee of Taizhou People's Hospital. Subjects who met the standards and gave informed consent to join the study were informed of the purpose of the study and signed informed consent. The study subjects were hospitalized patients in Taizhou People's Hospital from March 2022 to August 2022, including 4 patients in T2DM+MCI group and 4 patients in T2DM+nMCI group, Second-generation high-throughput sequencing was performed. There were 30 patients in T2DM+MCI group, and 30 patients in T2DM+nMCI group were verified by qRT-PCR.

Inclusion Criteria and Exclusion Criteria

The inclusion criteria of T2DM group were:: (1) Typical symptoms of DM + random blood glucose ≥11.1mmoL/L; (2) Oral Glucose Tolerance Test (OGTT) blood glucose ≥11.1mmoL/L for 2 hours; (3) Fasting blood glucose ≥7.0mmoL/L; (4) HbA1C≥6.5%. If one of the four criteria is met and the typical symptoms of DM (such as polydipsia, polyuria, hypereating, and weight loss) are present, T2DM is diagnosed. Patients with a clear history of T2DM were included in this study even if their blood glucose level failed to meet the diagnostic criteria for T2DM under the action of hypoglycemic drugs.

Excluded diagnostic criteria: (1) patients with acute complications such as diabetic ketoacidosis and acute infectious lesions; (2) Patients with severe hepatic and renal insufficiency; (3) Patients with other neuroendocrine diseases; (4) Patients with personal history of malignant tumor and mental illness; (5) Patients who have had weight-loss surgery in the past or are currently taking weight-loss medications; (6) Patients with chronic wasting diseases, anemia, and severe malnutrition; (7) Patients with cerebrovascular disease, brain trauma, stroke, intracranial tumor, history of craniocerebral surgery, toxic encephalopathy, Parkinson's disease, congenital mental impairment, epilepsy, major depression and other patients with serious impact on cognitive function evaluation; (8) Patients with hearing impairment and mental disorders; (9) Illiterate patients who refused to participate in the study.

MoCA Evaluation Basis and Grouping

MoCA scale is commonly used to assess cognitive function, mainly applicable to patients with high education level. Due to its high sensitivity, MoCA is widely used in clinical screening of cognitive function. MoCA involves eight test items including attention, memory, language expression ability and orientation, with a total score of 30 points. a score greater than 26 points is considered as normal cognitive function, and Less than 26 points is considered as cognitive impairment. When the age of education of the subject is less than 12 years, one point will be added. The following points should be noted in the scoring process: (1) Two professionally trained physicians from the research group should conduct the test. During the testing process, standardized language should be used to communicate with the tested subjects, and the MoCA score of the tested subjects should be the average of the two. (2) Select a quiet room, inform the patients of the purpose of the test, and obtain the understanding and support of the tested subjects; (3) If there are interference behaviors such as phone answering during the test, the test should be canceled and assessed at another time. According to MoCA scale, the patients were divided into two groups: T2DM+MCI group T2DM +nMCI group.

Experimental Reagents and Instruments (Table 1)