Avicenna Journal of Clinical Medicine

Volume 30, Issue 4

Original Article

Investigation of the Effectiveness of Doxycycline on the Improvement of Oxidative Stress Biomarkers in Patients with Ischemic Stroke

Mojtaba Khazaei1, Masrour Valdbeighi2, Salman Khazaei3, Efat Al-Sadat Mirmoeini4, Fereshteh Mehri5,*

  1. Department of Neurology, School of Medicine, Hamadan University of Medical Sciences, Hamadan, Iran

2 General Practitioner, Hamadan University of Medical Sciences, Hamadan, Iran

3 Autism Spectrum Disorders Research Center, Hamadan University of Medical Sciences, Hamadan, Iran

4 Occupational Health and Safety Research Center & Research Center for Health Sciences, Hamadan University of Medical Sciences, Hamadan, Iran

5 Nutritional Health Research Center & Occupational Health and Safety Research Center, Hamadan University of Medical Sciences, Hamadan, Iran

*Corresponding author: Fereshteh Mehri, Nutritional Health Research Center & Occupational Health and Safety Research Center, Hamadan University of Medical Sciences, Hamadan, Iran. Email: freshteh_mehri@yahoo.com

EXTENDED ABSTRACT

Background

Stroke is a major neurological cause of death and long-term disability and imposes substantial personal, family, and health-system costs [1]. World Health Organization data cited in the article indicate that approximately 15 million people experience stroke annually worldwide, with about 5 million deaths and another 5 million people left permanently disabled [2]. Ischemic stroke accounts for approximately 85% of all strokes [3]. Because the brain has high metabolic activity and limited energy reserves, interruption of cerebral blood flow rapidly deprives neural tissue of oxygen and nutrients and may cause irreversible cellular injury [4]. Although intravenous tissue plasminogen activator can reduce long-term complications in eligible patients, its use is limited by the narrow therapeutic window, contraindications, and cost [5]. Oxidative stress is an important component of ischemic brain injury. Excess reactive oxygen species (ROS) can damage neuronal structures and functions, while the brain is particularly vulnerable because of its high content of polyunsaturated fatty acids and relatively limited antioxidant defenses [6–10].

Doxycycline, a tetracycline derivative, has pharmacologic actions beyond its antibacterial activity, including anti-inflammatory, anti-apoptotic, and antioxidant effects [11]. Experimental evidence cited by the authors suggests that doxycycline can directly scavenge superoxide and inhibit formation of malondialdehyde (MDA) adducts, providing a plausible mechanism for attenuation of oxidative injury [12]. However, human clinical evidence in acute ischemic stroke remains limited. The present study therefore evaluated whether adding doxycycline to standard treatment improves serum biomarkers of oxidative stress and inflammation in patients with acute ischemic stroke.

Methods

This randomized clinical trial was conducted at Besat Hospital in Hamadan, Iran, in 2023 and included 48 patients with acute ischemic stroke who were admitted to the neurology ward within the first 24 hours after stroke onset. Eligible patients were 30–70 years old, had focal neurological deficits with a clinical diagnosis of acute hemispheric ischemic stroke, had MRI and CT findings compatible with the diagnosis, were not pregnant or breastfeeding, were experiencing a first ischemic stroke, and had not used antioxidant compounds during the preceding month. Exclusion criteria included acute or chronic intracerebral hemorrhage, cerebral aneurysm, nonischemic etiologies, cognitive or behavioral conditions preventing cooperation, withdrawal of consent, drug intolerance or adverse effects, a history of hypersensitivity to doxycycline or related products, and swallowing impairment requiring a nasogastric tube.

Participants were randomized equally to intervention and control groups using block allocation. An independent nurse who was not a member of the research team performed allocation. Both groups received standard stroke treatment. The intervention group additionally received doxycycline 100 mg every 12 hours for 7 days [13]. The control group received a placebo solution composed of starch and Avicel in water with an appearance and color similar to the active preparation. Serum samples were obtained before treatment and 7 days after the intervention. Ten milliliters of blood were collected from each participant; serum was separated and stored at -80 °C until analysis.

The assessed oxidative stress biomarkers were MDA, glutathione (GSH), total oxidant status (TOS), and total antioxidant capacity (TAC). TAC was assessed using the ferric reducing ability of plasma (FRAP) method, with absorbance measured at 593 nm [14]. TOS and antioxidant-related measures were also determined according to the relevant commercial kit procedures, and MDA was measured as an index of lipid peroxidation using thiobarbituric acid-reactive products with absorbance at 532 nm. Inflammatory biomarkers included C-reactive protein (CRP), measured using a semi-quantitative latex agglutination kit, and erythrocyte sedimentation rate (ESR), measured using an automated ESR reader. Data were analyzed with SPSS version 20. Quantitative variables were summarized as mean±standard deviation and categorical variables as frequency and percentage. Between-group comparisons used t tests or Mann–Whitney tests for quantitative variables and chi-square or Fisher exact tests for categorical variables, with P<0.05 considered statistically significant. The study was approved by the Ethics Committee of Hamadan University of Medical Sciences (IR.UMSHA.REC.1401.840) and registered in the Iranian Registry of Clinical Trials (IRCT20170114031921N6); written informed consent was obtained from all participants.

Results

All 48 patients were included in the analysis, with 24 participants in each group. The intervention and control groups did not differ significantly in baseline demographic characteristics, underlying diseases, or assessed risk factors. Before treatment, there were also no statistically significant between-group differences in MDA, TOS, TAC, GSH, CRP, or ESR.

Inflammatory indices decreased numerically in both groups but did not show statistically significant differences between groups. ESR changed from 11.9±11.9 to 6.1±3.0 in the intervention group and from 10.9±9.8 to 7.9±6.5 in the control group; the between-group P value after intervention was 0.610. CRP decreased from 4.6±2.5 to 3.5±3.1 with doxycycline and from 5.0±2.9 to 3.9±2.8 with placebo; the post-intervention between-group P value was 0.144.

Table 2. Comparison of inflammatory factors before and after intervention in both treatment groups.

After 7 days, oxidative stress markers changed in the same overall direction in both groups, but the differences between groups favored doxycycline. Mean MDA decreased from 152.9±80.5 to 54.4±55.2 in the intervention group and from 123.9±61.9 to 108.0±73.8 in the control group; the post-intervention difference was significant (P=0.001). Mean TOS decreased from 26.3±14.2 to 18.3±13.6 with doxycycline and from 27.5±12.9 to 25.5±8.4 with placebo, with a significant post-intervention between-group difference (P=0.013). Antioxidant indices increased after treatment. TAC rose from 4.4±1.7 to 5.5±2.4 in the intervention group and from 4.0±1.1 to 4.2±2.4 in the control group; the post-treatment difference was significant (P=0.025). GSH increased from 201.6±39.6 to 247.8±97.6 in the intervention group and from 191.3±47.9 to 204.5±86.7 in the control group, again with a significant post-intervention difference (P=0.048).

Table 3. Comparison of oxidative stress biomarkers before and after intervention in both treatment groups.

Thus, the principal treatment-associated findings were lower MDA and TOS and higher TAC and GSH after one week of adjunctive doxycycline, whereas CRP and ESR did not differ significantly between groups.

Conclusion

Adding doxycycline 100 mg every 12 hours for one week to standard treatment in patients with acute ischemic stroke was associated with improvement in serum oxidative stress and antioxidant biomarkers, including lower MDA and TOS and higher TAC and GSH, while inflammatory markers CRP and ESR did not differ significantly from placebo. These findings support a potential antioxidant effect of doxycycline in acute ischemic stroke. As stated by the authors, larger clinical studies are required to confirm the results and clarify the molecular mechanisms involved.

Keywords: Doxycycline, Ischemic Stroke, Oxidative Stress

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