Retrospective Cohort Study of Peripheral Blood miR-486-3p Levels in Patients with Acute Cerebral Infarction and Obstructive Sleep Apnea Syndrome

Summary

miR‑486‑3p is downregulated in acute cerebral infarction patients with obstructive sleep apnea syndrome, correlates negatively with disease severity and inflammatory markers, and predicts poor 1‑year prognosis, supporting its potential as a diagnostic and prognostic biomarker.

Abstract

Acute cerebral infarction (ACI) complicated with obstructive sleep apnea syndrome (OSAS) confers a high risk of stroke recurrence and death, yet effective biomarkers for this comorbid condition remain limited. This retrospective cohort study enrolled 270 patients with ACI between November 2020 and December 2023, who were classified into the ACI group (n = 100, apnea‑hypopnea index [AHI] < 5) and the ACI with OSAS group (n = 170, AHI ≥ 5), with the latter further stratified into mild (5 ≤ AHI < 15, n = 60), moderate (15 ≤ AHI < 30, n = 75), and severe (AHI ≥ 30, n = 35) subgroups. Serum miR‑486‑3p expression was measured by RT‑qPCR, and inflammatory markers, including tumor necrosis factor‑α (TNF‑α), C‑reactive protein (CRP), and hypoxia‑inducible factor‑1α (HIF‑1α), were quantified by ELISA. The diagnostic value of miR‑486‑3p was evaluated using receiver operating characteristic curve analysis, and its prognostic significance was assessed through Kaplan‑Meier survival analysis and Cox regression models. miR-486-3p was markedly lower in the comorbid group and demonstrated strong diagnostic ability (AUC = 0.927). Notably, a clear dose‑dependent trend was observed across OSAS severity subgroups, with miR‑486‑3p decreasing progressively and TNF‑α and CRP increasing stepwise from mild to severe OSAS. Spearman correlation analyses revealed that miR‑486‑3p was negatively correlated with AHI, NIHSS score, TNF‑α, and CRP. At 1‑year follow‑up, patients with low miR‑486‑3p expression had significantly worse functional outcomes compared with those with high expression. Multivariate Cox regression confirmed miR‑486‑3p as an independent protective factor for 1‑year prognosis. These findings suggest that miR‑486‑3p may serve as a novel diagnostic and prognostic biomarker for ACI patients with OSAS, potentially through its regulatory role in systemic inflammation, and warrant further validation in larger prospective cohorts.

Introduction

Obstructive sleep apnea syndrome (OSAS) is a common condition characterized by repeated interruptions in breathing during sleep¹. OSAS is often associated with comorbidities such as metabolic, cardiovascular, renal, pulmonary, and neuropsychiatric disorders². Beyond its general associations with cardiovascular and metabolic diseases, OSAS poses a specific and grave threat in acute ischemic stroke, complicating up to 70% of cases and increasing risks of neurological decline, mortality, and recurrent stroke^3,4,5. Acute cerebral in....

Protocol

This study was approved by the Ethics Committee of Affiliated Hospital of Xuzhou Medical University (approval number: XYFY2024-KL238-01) and was exempt from the requirement to obtain additional informed consent from patients. The reagents and the equipment used are listed in the Table of Materials.

Patients and specimens
This study was a retrospective cohort study. From November 2020 to December 2023, a total of 270 patients undergoing ACI from Affiliated Hospital of Xuzhou Medical University were consecutively recruited. The patient selection process is summarized in Supplementary Figure....

Results

Baseline characteristics in the ACI group and the ACI combined OSAS group
As shown in Table 1, the ACI group consisted of 68 males and 32 females, with an average age of 62.84 ± 12.65 years, and the ACI combined OSAS group included 116 males and 54 females, with an average age of 61.31 ± 11.25 years. The ACI combined OSAS group had significantly higher AHI, NIHSS scores, and CRP levels. However, the two groups showed comparable distributions of age, gender, BMI, hypertension, DM, car.......

Discussion

Patients with OSAS experience repeated airway collapse and obstruction, leading to intermittent hypoxia (IH) and excessive daytime sleepiness (EDS), which can cause neuronal damage¹⁴. OSA is highly prevalent after ACI, affecting approximately 72% of patients, and significantly worsens cardiovascular risk¹⁵. ACI constitutes a spectrum of neurological deficits arising from localized cerebral ischemia and hypoxia secondary to inadequate perfusion‌¹⁶

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
3.0T MRI Scanner	Siemens Healthineers, Germany	MAGNETOM Skyra	Brain imaging for acute cerebral infarction confirmation
96-well ELISA Microplates	Shenzhen Xinbosheng Biological Co., Ltd., China	XBS-PLATE	Antigen-antibody reaction for ELISA detection
AASM Clinical Practice Guidelines for Sleep-Disordered Breathing	American Academy of Sleep Medicine	2020 full guideline	Standardized AHI diagnostic and OSAS severity stratification criteria
Calgary Sleep Apnea Quality of Life Index (SAQLI) Chinese validated version	University of Calgary	2021 translated edition	Quality-of-life assessment for OSAS complicated ACI patients
EDTA-K2 5 mL Vacuum Blood Collection Tubes	BD Biosciences, USA	367863	Peripheral venous blood collection for serum separation
Epworth Sleepiness Scale (ESS) Chinese validated version	Johns Hopkins University	2018 revised edition	Evaluation of daytime sleepiness in stroke patients
GraphPad Prism 9	GraphPad Software, USA	Version 9.4.1	Scatter plots, boxplots, ROC curve visualization
Human CRP ELISA Kit	Shenzhen Xinbosheng Biological Co., Ltd., Shenzhen, China	XBS-H001	Quantitative detection of serum C-reactive protein
Human HIF-1α ELISA Kit	Shenzhen Xinbosheng Biological Co., Ltd., Shenzhen, China	XBS-H007	Quantitative detection of serum hypoxia-inducible factor-1α
Human TNF-α ELISA Kit	Shenzhen Xinbosheng Biological Co., Ltd., Shenzhen, China	XBS-H003	Quantitative detection of serum TNF-α inflammatory factor
IBM SPSS Statistics 23.0	IBM, Armonk, NY, USA	Version 23.0	All statistical analyses (t-test, correlation, ROC, Kaplan–Meier, Cox regression)
Microplate Spectrophotometer	Thermo Fisher Scientific (Waltham, MA, USA)	Multiskan FC	Absorbance reading for ELISA plates at 450 nm
miR-486-3p & U6 qPCR Primers	Sangon Biotech, Shanghai, China	Custom synthesis	Target and reference gene amplification for qPCR
miScript SYBR Green PCR Kit	Qiagen GmbH, Hilden, Germany	218073	Real-time quantitative PCR amplification of miR-486-3p and U6
NIH Stroke Scale (NIHSS) Chinese Validated Version	National Institute of Neurological Disorders and Stroke (NINDS), USA	Standard clinical edition	Baseline neurological severity assessment for ACI patients
PSG Sleep Scoring Software	BMC Medical Co., Ltd., China	v3.2	Automated preliminary scoring of sleep respiratory events
QuantStudio 6 Pro Real-Time PCR System	Applied Biosystems, Thermo Fisher Scientific (Waltham, MA, USA)	QuantStudio 6 Pro	RT-qPCR amplification, melting curve acquisition, Ct value recording
QuantStudio Real-Time PCR Software	Applied Biosystems, USA	v1.7	qPCR raw data export, Ct value analysis
Refrigerated Bench Centrifuge	Xiangyi Centrifuge Instrument Co., Ltd., Hunan, China	TDZ5-WS	Serum separation from whole blood (2700 × g, 4 °C)
RevertAid RT Reverse Transcription Kit	Thermo Fisher Scientific, Waltham, MA, USA	K1691	Reverse transcription of serum miRNA into cDNA
RNase-free 1.5 mL Microcentrifuge Tubes	Axygen, USA	MCT-150-C	Aliquoting and storage of separated serum
TRIzol RNA Isolation Reagent	Invitrogen, Thermo Fisher Scientific, Waltham, MA, USA	15596026	Total RNA extraction from human serum samples
Ultra-low Temperature Freezer	Haier Biomedical, Shandong, China	DW-86L388	Long-term serum sample storage at −80 °C
YH-600B Polysomnography Monitor	BMC Medical Co., Ltd., Beijing, China	YH-600B	Overnight sleep monitoring and AHI calculation for OSAS diagnosis