Study of Neuroinflammation in the Rat Hippocampus during Ethanol Exposure and Pharmacologic Correction with Azithromycin: New Data and Future Perspectives

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Abstract

With prolonged ethanol ingestion, disturbances in the emotional spectrum develop, and memory problems are noted. These symptoms may be mediated by the development of neurochemical changes in the hippocampus of the brain. Although there is evidence that the hippocampus is vulnerable to chronic alcohol intoxication and the development of neuroinflammation and neurodegeneration in this brain region, the key molecular mechanisms have not been identified. The aim of the study was to investigate changes in the immune system in the periphery as well as in the hippocampus of the rat brain during ethanol exposure and during pharmacological correction with azithromycin (AZM). Long-term ethanol exposure was modeled by injecting rats with a 20% ethanol solution (4 g/kg) for 4 weeks. General biochemical and clinical blood analysis was performed in animals. The expression levels of cytokine genes (Il1β, Ccl2, Il6, Il11, Il13, Tnfα, Tgfβ) and toll-like receptor system genes (Tlr3, Tl4, Tlr7, Nfkb1, Hmgb1) and TLR system-related microRNA molecules (miR-182, miR-155-5p, miR-96-5p, miR-let-7b) were evaluated in the hippocampus. IL-1β protein content was also assessed in the hippocampus. Prolonged alcoholization caused an increase in mRNA and protein levels of IL-1β, and a decrease in mRNA levels of Tnfα, Il11, Tlr3, and Tlr7. The contents of miR-let-7b, miR-96, and miR-155 were downregulated in the hippocampus during long-term alcoholization. Elevated Il1β mRNA and protein levels and Hmgb1 mRNA levels were maintained under conditions of ethanol withdrawal. Tlr3 mRNA levels were decreased under withdrawal. Administration of AZM reduced IL-1β, TLR3 and HMGB1 mRNA levels under conditions of ethanol withdrawal, and at higher doses of the drug, a decrease in IL-1β protein levels in the hippocampus of rat brain was observed. Thus, the study provided new insights into the mechanisms of neuroinflammation in the brain hippocampus during prolonged exposure to ethanol and upon withdrawal. The results obtained allowed us to outline a number of tasks for further studies in this direction.

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About the authors

M. I. Airapetov

Institute of Experimental Medicine; Kirov Military Medical Academy

Author for correspondence.
Email: interleukin1b@gmail.com
Russian Federation, 197376, Saint Petersburg; 194044, Saint Petersburg

S. O. Eresko

Institute of Experimental Medicine; Saint Petersburg National Research University of Information Technologies, Mechanics and Optics

Email: interleukin1b@gmail.com
Russian Federation, 197376, Saint Petersburg; 197022, Saint Petersburg

S. A. Shamaeva

Institute of Experimental Medicine

Email: interleukin1b@gmail.com
Russian Federation, 197376, Saint Petersburg

E. R. Bychkov

Institute of Experimental Medicine

Email: interleukin1b@gmail.com
Russian Federation, 197376, Saint Petersburg

A. A. Lebedev

Institute of Experimental Medicine

Email: interleukin1b@gmail.com
Russian Federation, 197376, Saint Petersburg

P. D. Shabanov

Institute of Experimental Medicine; Kirov Military Medical Academy

Email: interleukin1b@gmail.com
Russian Federation, 197376, Saint Petersburg; 194044, Saint Petersburg

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2. Fig. 1. Cytokine mRNA content in the brain hippocampus of rats with long-term ethanol consumption; * p ≤ 0.05 - relative to the control group

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3. Fig. 2. Relative level of mRNA content of TLR system genes in the hippocampus of the brain of rats with long-term ethanol consumption; * p ≤ 0.05 - in relation to the control group

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4. Fig. 3. The mRNA content of cytokines and TLR system genes in the brain hippocampus of rats during ethanol withdrawal; * p ≤ 0.05 - in relation to the control group

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5. Fig. 4. The mRNA content of TLR system genes in rat brain hippocampus under pharmacological correction with azithromycin under conditions of ethanol withdrawal; * p < 0.05 - in relation to the control group; # p < 0.05 - in relation to the ‘ethanol + water withdrawal’ group

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6. Fig. 5. Cytokine mRNA content in rat brain hippocampus during pharmacological correction with azithromycin under conditions of ethanol withdrawal; * p < 0.05 - in relation to the control group; # p < 0.05 - in relation to the ‘ethanol + water withdrawal’ group

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7. Fig. 6. IL-1β protein content in the hippocampus of rat brain; * p < 0.05 - relative to the control group; # p < 0.05 - relative to the ‘ethanol + water cancellation’ group

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