Brain damage in photoinduced ischemia under streptozotocin-induced diabetes in mice with pannexin 1 protein gene knockout

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Abstract

Diabetes mellitus (DM), along with ischemia, is one of the top ten causes of death in the globalpopulation, according to the latest World Health Organization (WHO) data. Clinical research data have revealed a high risk of stroke and heart attacks in patients with diabetes. However, there is still a lack of understanding of the involvement of pannexin 1 (Panx1) protein in cerebral ischemia combined with DM. In the presented study, we used the Panx1 gene knockout mice in models of streptozotocin-induced diabetes and photoinduced ischemia to investigate the effect of the Panx1 on the severity of ischemic brain damage and systemic inflammation in mice with a combination of cerebral ischemia and diabetes. It has been found that under conditions of experimental diabetes, the knockout of the Panx1 gene significantly reduces the size of the ischemic lesion, stabilizes the ischemia-induced increase in the blood-brain barrier permeability, reduces the number of errors in the sensorimotor test and the level of neutrophils in the blood. It is important to note that the Panx1 knockout exhibits a protective effect only in the presence of diabetes mellitus, without significantly affecting the severity of ischemic brain injury in mice without streptozotocin-induced diabetes. Panx1 knockout also did not affect the severity of hyperglycemia in animals in this diabetes model. It can be assumed that the efficacy of the treatment for pathologies combined with diabetes mellitus can be enhanced by incorporating pannexin channel blockers into the complex therapy, representing a novel approach to addressing these serious conditions.

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About the authors

М. А. Nikolaenko

Lomonosov Moscow State University

Email: gorbi67@mail.ru
Russian Federation, Moscow

М. V. Gulyaev

Lomonosov Moscow State University

Email: gorbi67@mail.ru
Russian Federation, Moscow

А. А. Volkova

Lomonosov Moscow State University; Pirogov Russian National Research Medical University

Email: gorbi67@mail.ru
Russian Federation, Moscow; Moscow

L. R. Gorbacheva

Lomonosov Moscow State University; Pirogov Russian National Research Medical University

Author for correspondence.
Email: gorbi67@mail.ru
Russian Federation, Moscow; Moscow

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Supplementary files

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2. Fig. 1. Dynamics of changes in blood glucose levels in mice of the studied groups. WT – wild type (without diabetes), Panx 1 KO – pannexin 1 gene knockout (without diabetes), WTd, KOd – corresponding groups of mice with experimental diabetes. WT n = 7, WTd n = 9, KO n = 18, KOd n = 15. Data are presented as means ± standard deviation. Three-way ANOVA, post hoc Šidák test, differences are significant: ** – p < 0.01, # – p < 0.0001 (compared with the same group on the 1st day of the experiment).

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3. Fig. 2. Change in body weight in mice of the studied groups on the last day of the experiment compared to the first day. WT – wild type (without diabetes, n = 6), KO – pannexin 1 gene knockout (without diabetes, n = 17); WTd (n = 9), KOd (n = 15) – corresponding groups of mice with diabetes mellitus. Data are presented as means ± standard deviation. Three-way ANOVA, post hoc Šidák test, differences are significant: * – p < 0.05, *** – p < 0.001.

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4. Fig. 3. Photo-induced ischemia-induced brain injury in wild-type and diabetic Panx1 knockout mice. (a) Volume of ischemic lesion after photothrombosis in wild-type and diabetic Panx1 knockout mice (data normalized to mean values ​​in corresponding control groups, non-diabetic animals (control = 1)), (b) T2-weighted MRI images of brain injury in the studied animal groups. WT n = 11, WTd n = 7, KO n = 21, KOd n = 13. Differences are significant: ** – p < 0.01.

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5. Fig. 4. Evaluation of Evans blue (EB) extravasation and BBB permeability after photoinduced ischemia in wild-type (WTd) and Panx1 knockout (KOd) mice with diabetes. (a) – comparison of EB extravasation in the studied groups of animals (WT n = 5, WTd n = 8, KO n = 16, KOd n = 14), (b) – comparison of BBB permeability in the studied groups of animals (WT n = 9, WTd n = 4, KO n = 21, KOd n = 9), (c) – visual photographs of the brain after photothrombosis, EB administration a day later and perfusion on the second day after ischemic injury in mice. The data are normalized to the mean values ​​in the corresponding control groups, in animals without diabetes (control = 1). The differences are significant: ** – p < 0.01, *** – p < 0.001.

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6. Fig. 5. Sensorimotor status of animals in the study groups assessed using the “Grate” test. Change in the proportion (in percent) of motor errors per 100 steps of the animal after FT compared to the values ​​before FT in wild-type mice (WTd, n = 8) and mice with the Panx1 gene knockout (KOd, n = 13) against the background of diabetes. WT n = 5, KO n = 16. Data are normalized to the mean values ​​in the corresponding control groups, in animals without diabetes (control = 1). Differences are reliable: *** – p < 0.001; # – p < 0.0001.

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7. Fig. 6. Evaluation of neutrophil levels in wild-type (WTd) and Panx1 knockout (KOd) mice with diabetes. (a) comparison of neutrophil counts in diabetic experimental groups before photothrombosis (WT n = 5, WTd n = 9, KO n = 17, KOd n = 15), (b) change in neutrophil counts on the last day of the experiment compared to the first day (24 h after photothrombosis with diabetes, WT n = 6, WTd n = 9, KO n = 16, KOd n = 13). Data are normalized to mean values ​​in the corresponding control groups, in animals without diabetes (control = 1). Differences are significant: ** – p < 0.01.

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