Resident macrophages of different organs have structural and functional features, which can complicate their identification and analysis. A promising candidate for the role of a universal immunohistochemical marker of resident macrophages is the calcium-binding protein Iba-1, a well-known marker of brain microglia. The purpose of this work was to study the possibility of using one variant of antibodies to the Iba-1 protein for the immunohistochemical detection of resident macrophages in the liver, myocardium, lung, and choroid plexus of the rat brain. The study was performed on male Wistar rats (n = 15). It was shown that the use of rabbit monoclonal antibodies against Iba-1 allows highly effective detection of Kupffer cells in the liver, resident macrophages in the myocardium, alveolar and interstitial macrophages in the lung, and Kolmer cells in the choroid plexus of the rat brain. In all cases, the reaction is characterized by a high specificity and the absence of background staining. In contrast to the classical marker of macrophages, the CD68 molecule, the Iba-1 protein is evenly distributed in the cytoplasm of cell bodies and processes. This makes it possible to more fully identify cells using immunostaining for Iba-1, carry out their three-dimensional reconstructions, and study their structural and functional organization. Immunohistochemical reaction against Iba-1 can be successfully used as a universal alternative to other common methods for identifying resident macrophages.

BACKGROUND: The choroid plexus of the brain is the source of cerebrospinal fluid and a major component of the blood–cerebrospinal fluid barrier, providing active transport of only essential substances and preventing the entry of harmful substances, including proinflammatory molecules, pathogens, and toxins. Basement membranes of the choroid plexus play a special role in the implementation of barrier functions, which underlie the choroidal epithelium and capillary endothelium, and serve as an additional filter for substances penetrating from the blood into the cerebrospinal fluid. The morphological organization of basement membranes in the villi of the human choroid plexus has not been examined extensively. AIM: to analyze the organization of basement membranes in the villi of the human telencephalon choroid plexus by immunohistochemical detection of type IV collagen. MATERIALS AND METHODS: The study was performed on archival materials from the choroid plexus of the human brain (n=10; age 29–50 years) using immunohistochemical methods for detecting type IV collagen. RESULTS: An immunohistochemical reaction using antibodies to type IV collagen showed the distribution of this protein in the subepithelial area and stroma of the choroid plexus villi. All immunopositive structures had clear contours. No reaction in the cell cytoplasm or a nonspecific background was noted. Contacts of subepithelial and subcapillary basement membranes labeled with antibodies to type IV collagen were not detected. CONCLUSION: The results showed different organization of the basement membranes of the villi of the choroid plexus of the human brain in the subepithelial and perivascular areas. In this case, the subepithelial and perivascular components containing type IV collagen did not merge.

Background: Acute and chronic brain damage in type 2 diabetes mellitus (DM) determines the need to investigate the neuroprotective potential of glucose-lowering drugs. The purpose was to directly compare the neuroprotective effects of glucagon-like peptide-1 receptor agonists (GLP-1RAs) with different duration of action and sodium-glucose cotransporter-2 inhibitors (SGLT-2i) in type 2 diabetic rats with and without stroke. Methods: DM was modelled using high-fat diet and nicotinamide+streptozotocin protocol. The following groups (n = 15 each) were formed: DM without treatment, treatment with liraglutide, dulaglutide, canagliflozin as well as control group without DM and treatment. After 8 weeks, 10 rats from each group underwent middle cerebral artery occlusion. In the reperfusion period neurological deficit, neuroglial damage markers and brain necrosis were evaluated. Brain slices from the remaining 5 animals in each group were histologically examined for microglial activation and neuronal damage. Results: Brain damage was similar in “DM” and “Control” (17.53 [14.23; 26.58] and 15.87 [13.40; 22.68] % of total brain volume, respectively). All study drugs diminished damage volume comparing with “DM” and “Control” whereas the necrosis volume in “DM+Liraglutide” was smaller than in “DM+Canagliflozin” and did not significantly differ from “DM+Dulaglutide” (2.9 [1.83; 4.71], 6.17 [3.88; 8.88] and 4.57 [3.27; 7.90] %). The neurological deficit was more prominent in “DM” than in “Control”, while all the drugs demonstrated similar positive effect. Neurofilament light chains (NLC) did not differ between “DM” and “Control”. Dulaglutide and canagliflozin caused a marked decrease in NLC. Protein S100BB level was similar in “DM” and “Control”. Liraglutide caused the largest S100BB decrease, while canagliflozin did not influence it. In chronic brain ischaemia, all drugs increased the number of normal neurons, but GLP-1RAs had a more pronounced effect. DM was accompanied by increased number of activated microglial cells in Cornu Ammonis (CA)1 hippocampal region. Both GLP-1RAs reduced the number of Iba-1-positive cells, with dulaglutide being more effective than liraglutide, whereas canagliflozin did not affect this parameter. Conclusions: GLP-1RAs and SGLT-2i have neuroprotective properties against acute and chronic brain damage in diabetic rats, although the infarct-limiting effect of GLP-1RAs may be more pronounced. GLP-1RAs and SGLT-2i exert their protective effects by directly influencing neuronal survival, whereas GLP-1RAs also affect microglia.

Belok Iba-1 tradicionno schitayut vysokoselektivnym markerom mikroglii blagodarya specificheskoj ekspressii gena imenno v etoj populyacii kletok CNS. V rezul'tate al'ternativnogo splajsinga formiruetsya neskol'ko izoform belka Iba-1, chto mozhet sluzhit' prichinoj raskhozhdenij rezul'tatov immunogistohomicheskih reakcij v zavisimosti ot togo, k kakim epitopam immunogena vyrabotany vybrannye dlya issledovaniya antitela. V svyazi s etim, a takzhe s neobhodimost'yu opredeleniya nadezhnyh variantov antitel k Iba-1, dostupnyh dlya issledovatelej v Rossijskoj Federacii, cel'yu raboty bylo ocenit' rezul'taty vyyavleniya mikroglii i makrofagov s ispol'zovaniem antitel k razlichnym posledovatel'nostyam belka, vypuskaemyh raznymi proizvoditelyami. Materialom dlya issledovaniya sluzhili obrazcy golovnogo mozga i semennika polovozrelyh (3–5 mesyacev) krys-samcov Wistar (n = 8). V kachestve pervichnyh reagentov ispol'zovali poliklonal'nye i monoklonal'nye (klon JM36-62) antitela k Iba-1. Ustanovleno, chto monoklonal'nye antitela klona JM36-62 pozvolyayut dobit'sya bolee vysokoj selektivnosti vyyavleniya antigena pri luchshem sootnoshenii signal/fon i prigodny dlya zameny reagentov, ne dostupnyh v nastoyashchee vremya k priobreteniyu. Ispol'zovanie poliklonal'nyh antitel privelo ne tol'ko k immunospecifichnoj vizualizacii mikroglii i makrofagov, no i k vyyavleniyu kletok epitelio-spermatogennogo sloya semennika. Predpolagaetsya, chto v kletkah epitelio-spermatogennogo sloya prisutstvuet izoforma Iba-1, lishennaya epitopa, sootvetstvuyushchego posledovatel'nosti immunogennogo dlya antitel klona JM36-62 fragmenta nativnogo belka. Funkcional'noe znachenie razlichnyh izoform Iba-1 na nastoyashchij moment ostaetsya neyasnym i trebuet dal'nejshih issledovanij.

BACKGROUND: Chronic brain dyscirculation is one of the frequent type 2 diabetes mellitus (DM) complications and leads to patients’ disability. Sodium-glucose co-transporter type 2 inhibitors (SGLT-2i) have been proven to have advantages for cardiovascular system, but their effect on the central nervous system (CNS) has not been studied enough.AIM: To study empagliflozin effect on CNS damage functional and laboratory parameters in patients with type 2 DM and, under experimental conditions, to investigate the mechanisms of the drug neurotropic effect.MATERIALS AND METHODS: The clinical part of the study included patients with type 2 DM on metformin monotherapy (n=39). Patients with a target glycated hemoglobin level formed the “MET” group (n=19), in patients with a non-target glycated hemoglobin level empagliflozin was co-administered for the following 6 months (the “MET+EMPA” group, n=20). Healthy volunteers comprised the control group (n=16). The cognitive status and neuron-specific enolase (NSE) and neurofilament light chains (NLC) concentration were studied. DM was modeled in rats, thereafter the rats were treated with empagliflozin for 8 weeks. Microglia activation was assessed using anti-Iba-1 antibodies and morphological changes in neurons when stained by the Nissl method.RESULTS: Both in the “MET+EMPA” and the “MET” groups cognitive deficits were observed, according to the Montreal Cognitive Assessment (MOCA) (24.0 (23.0; 27.0) and 25.0 (21.0; 27.0) points) and the Mini-Mental State Examination (MMSE) (23.75 (23.0; 27.0) and 25.0 (21.0; 27.0) points). Empagliflozin therapy led to the cognitive status normalization after 6 months (26.5 (24.0; 27.0) points according to the MOCA scale and 27.5 (24.0; 28.0) points according to the MMSE). Initially, all patients had a significant increase of NSE (3.60 (2.66; 3.76) ng/ml in the “MET” group, 3.22 (2.94; 3.54) ng/ml in the “MET+EMPA» group, 2.72 (2.13; 2.72) ng/ml in the «Control» group) and NLC (4.50 (3.31; 5.56) ng/ml in the «MET» group, 5, 25 (3.75; 6.25) ng/ml in the «MET+EMPA» group comparing with 3.50 (2.25; 3.50) ng/ml in the «Control» group). Empagliflozin therapy led to a significant decrease in NLC already after 3 months (3.80 (3.25; 3.87) ng/ml), without significant influence on the NSE level. In the experiment, DM was characterized by an increased number of activated microgliocytes and destructured neurons and a decreased number of neurons with a normal structure. Empagliflozin therapy was accompanied by a decrease in the number of immunopositive microgliocytes in the CA1 zone of the hippocampus and an increase in the number of structured neurons.CONCLUSION: Type 2 diabetes mellitus is characterized by functional and biochemical changes in the central nervous system even under satisfactory glycemic control. Therapy with empagliflozin has a neuroprotective effect, manifested in an improvement in cognitive status and a decrease in NLC level. Empagliflozin reduces neuronal damage and abnormal microglial activation.

Background: The accumulation of specific IgG antibodies in blood serum is considered a key criterion for the effectiveness of vaccination. For several vaccine-preventable infections, quantitative indicators of the humoral response have been established, which, when reached, provide a high probability of protection against infection. The presence of such a formal correlate of vaccine effectiveness is crucial, for example, in organizing preventive measures and validating newly developed vaccines. However, can effective protection against infection occur when the level of serum antibodies is lower than that provided by parenteral vaccination? Will protection be sufficient if the same vaccine antigen is administered via mucosal membranes without achieving high levels of specific IgG circulating in the blood? Methods: In this study, we compared the immunogenicity and protective efficacy of parenteral and mucosal forms of vaccines in experimental animals, targeting infections caused by the SARS-CoV-2 coronavirus and Streptococcus pneumoniae. We investigated the protective properties of a fragment of the coronavirus S1 protein administered intramuscularly with an adjuvant and orally as part of the probiotic strain Enterococcus faecium L3 in a Syrian hamster model. A comparative assessment of the immunogenicity and protective efficacy of a recombinant tandem (PSP) of immunogenic peptides from S. pneumoniae surface proteins, administered either parenterally or orally, was performed in a Balb/c mouse model. Results: Both models demonstrated significant differences in the immunogenicity of parenteral and oral vaccine antigens, but comparable protective efficacy.

The Iba-1 protein is traditionally considered a highly selective marker of microglia because of the specific expression of the gene in this particular population of the CNS cells. Alternative splicing creates several isoforms of the Iba-1 protein, which may cause discrepancies in the results of immunohistochomic reactions depending on which epitopes of the immunogen the antibodies selected for the study were developed. In this connection, and with the aim at identifying reliable variants of antibodies to Iba-1 available to researchers in the Russian Federation, we organized with study, seeking to evaluate the results of detecting microglia and macrophages using antibodies to different protein sequences produced by different manufacturers. As material, we used samples of the brain and testis of sexually mature (3–5 months) male Wistar rats (n = 8). Polyclonal and monoclonal (clone JM36-62) antibodies to Iba-1 were used as primary reagents. We found that monoclonal antibodies of the JM36-62 clone enable more selective antigen detection with a better signal/background ratio; they can be used as replacements for reagents that are currently not available commercially. Polyclonal antibodies enabled not only immunospecific imaging of microglia and macrophages, but also the identification of cells of the epithelial-spermatogenic layer of the testis. It is assumed that epithelial-spermatogenic layer contains the Iba-1 isoform devoid of an epitope that corresponds to the sequence of the immunogenic antibody clone JM36-62 fragment of the native protein. Functionally, various isoforms of Iba-1 should be investigated further.

BACKGROUND: Resident macrophages of the myocardium are necessary for the normal development and functioning of the cardiovascular system and play an important role in the pathogenesis of a number of cardiovascular diseases. AIM: To develop a universal methodological approach for the immunohistochemical identification of macrophages in the human and rat myocardium. MATERIAL AND METHODS: For the study, samples of the myocardium of people of both sexes aged from 28 to 52 years (n=7) and Wistar female rats (at the age of 15 months, (n=5)) were used. The work was performed using immunohistochemical staining method. RESULTS: The possibility of using polyclonal antibodies against the calcium-binding protein Iba1, which is a well-known microglial marker marker, for the detection of macrophages in the myocardium was shown. An optimal drug processing protocol was developed, the use of which makes it possible to equally effectively detect macrophages in the human and rat myocardium using only one antibody variant. CONCLUSION: A universal methodological approach has been proposed for the identification of macrophages in the human and rat myocardium, which can be successfully used in scientific and clinical diagnostic studies.

Academician A.A. Zavarzin (18861945) the founder of evolutionary histology and the creator of tissue parallelism theory, is known as an outstanding neurohistologist and organizer of the Department of General Morphology of the All-Union Institute of Experimental Medicine (AIEM). During the Great Patriotic War (World War II) A.A. Zavarzin was evacuated to Tomsk as part of the laboratories of the central part of AIEM (Moscow) and the Leningrad branch of AIEM (Leningrad). Some information about his scientific work in Tomsk is presented in the historical literature, however, up to the present, his organizational activities as a commissioner have not been known. Based on information from the archive of the Institute of Experimental Medicine (St. Petersburg), including previously unpublished materials from orders on personnel for 1941 and 1945 and those for the Tomsk Group of AIEM, this article presents new facts about the life and scientific and organizational activities of Academician A.A. Zavarzin during the Great Patriotic War. In the period from 1943 to 1944, he headed the Tomsk group of laboratories of the A.M. Gorky All-Union Institute of Experimental Medicine the main scientific medical institution in the country, on the basis of which the USSR Academy of Medical Sciences was organized in 1945.

BACKGROUND: A necessary attribute of any study, which is related to the cell biology of various structural components of the digestive tract, is the usage of modern immunohistochemical methods. One of the most difficult objects are resident liver macrophages, or Kupffer cells. This determines the high significance to develop a reliable approach for Kupffer cells visualization. AIM: The aim of the study was to develop a protocol for the immunohistochemical research of Kupffer cells in rat liver using two types of primary antibodies against Iba-1, and to analyze the advantages and disadvantages of this approach, taking into account the use of zinc-ethanol-formaldehyde as a fixative. MATERIALS AND METHODS: The study was carried out on liver samples of adult Wistar rats (n = 5). Goat polyclonal and rabbit monoclonal antibodies against calcium-binding protein Iba-1 were used for the ligh microscopy immunohistochemistry assay of resident liver macrophages. RESULTS: Using two types of antibodies it was shown by quantitative analysis that rabbit monoclonal antibodies most completely reveal Iba-1-immunopositive structures compared to goat polyclonal antibodies. Fixation with zinc-ethanol-formaldehyde made it possible to reveal Iba-1-immunopositive cells in all studied rat liver samples. CONCLUSIONS: Iba-1 immunohistochemistry using rabbit monoclonal antibodies was considered as the most optimal immunohistochemical approach. Fixation with zinc-ethanol-formaldehyde preserves the antigenic epitopes and allows the effective use of different antibodies.

Objective: Immunohistochemical investigation of archival histological material is a serious problem, since long-term storage of biological tissues, most often in formalin, leads to a loss of antigenic properties. However, the biological material can also be stored in the clearing agent methyl salicylate. The aim of this study was to assess the antigenicity of the human choroid plexus after extra long-term storage in methyl salicylate. Material and Method: The study was performed on samples of fixed human choroid plexus (occasionally with attached neighboring pineal gland) stored in either methyl salicylate or paraffin blocks for 25 years. Chromogenic and fluorescence immunohistochemistry of vimentin, GFAP, type IV collagen, β-catenin, α-smooth muscle actin, von Willebrand factor, CD68, mast cell tryptase, TMEM119, and synaptophysin was carried out. Results: The storage of human choroid plexus in methyl salicylate for 25 years does not impair its histomorphology and preserves the properties of all the antigens assessed, which makes their immunohistochemical visualization possible using both light and fluorescence microscopy. Additionally, we found that long-term storage of human choroid plexus in methyl salicylate does not cause an increase in autofluorescence. Conclusion: Methyl salicylate can be recommended as a medium for long-term storage of biological tissue, as it provides excellent brain tissue preservation and retains its antigenic properties for up to 25 years.

The work aimed to investigate the biocompatibility and biological activity of the water-soluble fullerene adduct C60-Arg. It was found that the material is haemocompatible, is not cyto- and genotoxic, possesses pronounced antioxidant activity. Additionally, this paper outlines the direction of application of water-soluble fullerene adducts in the creation of neuroprotectors. It has been suggested that a putative mechanism of the protective action of the C60-Arg adduct is associated with its antioxidant properties, the ability to penetrate the blood-brain barrier, and release nitrogen monoxide as a result of the catabolism of L-arginine residues, which promote vascular relaxation. The action of the C60-Arg adduct was compared with the action of such an antioxidant as Edaravone, which is approved in Japan for the treatment of ischemic and haemorrhagic strokes.

The declaration of the conclusion of the COVID-19 pandemic notwithstanding, coronavirus remains prevalent in circulation, and the potential emergence of novel variants of concern introduces the possibility of new outbreaks. Moreover, it is not clear how quickly and to what extent the effectiveness of vaccination will decline as the virus continues to mutate. One possible solution to combat the rapidly mutating coronavirus is the creation of safe vaccine platforms that can be rapidly adapted to deliver new, specific antigens in response to viral mutations. Recombinant probiotic microorganisms that can produce viral antigens by inserting specific viral DNA fragments into their genome show promise as a platform and vector for mucosal vaccine antigen delivery. The authors of this study have developed a convenient and universal technique for inserting the DNA sequences of pathogenic bacteria and viruses into the gene that encodes the pili protein of the probiotic strain E. faecium L3. The paper presents data on the immunogenic properties of two E. faecium L3 vaccine strains, which produce two different fragments of the coronavirus S1 protein, and provides an assessment of the protective efficacy of these oral vaccines against coronavirus infection in Syrian hamsters.

The COVID-19 pandemic was declared in March 2020 and is still ongoing. The course of the epidemic remains uncertain, and mutations in the coronavirus, as well as the emergence of new variants of concern, make it possible that there will be periodic surges in the number of cases. Moreover, it is not clear how quickly and to what extent the effectiveness of vaccination will decline as the virus continues to mutate. One possible solution to combat the rapidly mutating coronavirus is the development of pan-coronavirus vaccines that stimulate the production of antibodies against conserved elements of the virus. Another approach could be the creation of safe vaccine platforms that can be rapidly adapted to deliver new, specific antigens in response to viral mutations. Recombinant probiotic microorganisms that can produce viral antigens by inserting specific viral DNA fragments into their genome show promise as a platform and vector for mucosal vaccine antigen delivery. With a well-established safety profile and the ability to transform into vaccine candidates, probiotic bacteria have significant research potential. The authors of this study have developed a convenient and universal technique for inserting the DNA sequences of pathogenic bacteria and viruses into the gene that encodes the pili protein of the probiotic strain E. faecium L3. The paper presents data on the immunogenic properties of two E. faecium L3 vaccine strains, which produce two different fragments of the coronavirus S1 protein, and provides an assessment of the protective efficacy of these oral vaccines against coronavirus infection in Syrian hamsters.

NeuN is a neuron-specific nuclear protein expressed in most mature neuronal cell types, with some exceptions. These exceptions are known mainly for the brain but not for the spinal cord or the spinal visceral networks for which only scarce information is available. One of the most defined visceral structures in the spinal cord is the sympathetic intermediolateral nucleus located within the thoracolumbar segments. We investigated the NeuN staining in the intermediolateral nucleus and compared it with the staining for two neurochemical markers of visceral neurons: nitric oxide synthase and calcium-binding protein calretinin in adult cats and in kittens aged 0, 14, and 35 days. A clear NeuN-immunonegativity was obtained for intermediolateral neurons labeled for nitric oxide synthase for both adult cats and kittens. In contrast, a matched immunopositivity for the NeuN and calretinin was obtained, showing an age-dependent degree of this colocalization, which was high in newborn kittens, decreased on postnatal 14 and 35 days and persisted at a moderate level up to adulthood. Perhaps our data displayed a heterogeneity of the intermediolateral neurons.

We studied the reaction of rat hippocampal microgliocytes to hyperbaric oxygen at a pressure of 5 ata (absolute atmosphere). Immunohistochemical analysis with selective macrophage marker CD68 (ED1) and microglial marker Iba-1 allowed separate analysis of these two cell populations. It was shown that macrophages do not significantly contribute to reactive changes in the total pool of Iba-1+ hippocampal cells induced by hyperbaric oxygen.

Pulmonary embolism is a life-threatening condition, which can result in respiratory insufficiency and death. Blood clots occluding branches of the pulmonary artery (PA) are traditionally considered to originate from thrombi in deep veins (usually in legs). However, growing evidence suggests that occlusion of the vessels in the lungs can develop without preceding deep vein thrombosis (DVT). In this work, we used an inferior vena cava (IVC) complete ligation model of DVT in Wistar rats to explore the possibility and mechanisms of PA thrombosis under the conditions where all routes of thrombotic mass migration from peripheral veins are blocked. We demonstrate that rats both with normal and reduced neutrophil counts developed thrombi in the IVC, although, neutropenia caused a substantial decrease in thrombus size and a shift from fresh fibrin toward mature fibrin and connective tissue inside the thrombus. Massive fibrin deposition was found in the PA branches in the majority of DVT rats with normal neutrophil counts, but in none of the neutropenic animals. Neutrophil ablation also abolished macroscopic signs of lung damage. Altogether, the results demonstrate that thrombi in the lung vasculature can form in situ by mechanisms that require local neutrophil recruitment taking place in the DVT setting.

Hyperbaric oxygen (HBO2) causes a toxic effect on the central nervous system via poorly understood mechanisms. The aim of this study was to test the hypothesis that HBO2 inhibits the activity of glutamine synthetase (GS) in the brain and increases the level of glutamatergic neurotransmission, thus leading to the development of epileptiform activity in the EEG and tonic-clonic motor seizures. In experiments on awake rats exposed to oxygen partial pressure of 4.65 ATA, the seizure onset latency was determined in animals with intact and inhibited GS. The enzymatic activity of GS was irreversibly suppressed using the selective inhibitor L-methionine sulfoximine (MSO). It was found that the development of seizures under hyperbaric hyperoxia correlated with a decrease in the GS level in the brain. MSO accelerated the development of oxygen-induced seizures. The obtained data indicate that the manifestation of convulsive syndrome under extreme hyperoxia depends on the enzymatic activity of GS, a key enzyme regulating glutamatergic neurotransmission.

Sodium–glucose cotransporter type 2 inhibitors (SGLT2i) have proven cardioprotective properties, which makes this class of drugs one of the priorities in the treatment of patients with type 2 diabetes mellitus (DM2). Ischemic stroke and chronic cerebral dyscirculation occur with high frequency in DM2, thus actualizing a study of SGLT2i neurotropic properties. The aim of our study was to evaluate and compare the neuroprotective effects of the highly selective SGLT2i empagliflozin (EMPA) and low-selective SGLT2i canagliflozin (CANA) on the rat model of acute cerebral ischemia, as well as investigate the probable mechanism of these effects on the brain. At the first stage, EMPA and CANA were administered to Wistar rats without DM2 for 7 days prior to transient focal 30-min cerebral ischemia modeling. After 48 h of reperfusion, neurological deficit was assessed by Garcia scores, and brain sections were then incubated in a triphenyltetrazolium chloride solution to evaluate the brain damage volume. The latter did not differ in EMPA and CANA groups and was significantly smaller compared to the control group of untreated rats. At the same time, neither EMPA nor CANA had a significant effect on neurological deficit. At the second stage, we modeled DM2 (high-fat diet and streptozotocin + nicotinamide), and 4 weeks later, 8-week EMPA and CANA therapy was initiated. After the end of therapy, brain tissue was studied immunohistochemically. DM2 development was accompanied by an increase in the number of microgliocytes in the hippocampal CA1 area; the therapy with EMPA, but not CANA, led to a decrease in the number of activated microgliocytes. Thus, the highly selective SGLT-2i EMPA and low-selective SGLT-2i CANA exert a similar infarct-limiting effect when applied for 7 days prior to ischemia modeling in Wistar rats without DM2. The neuroprotective effect of EMPA in DM2 may in part be due to a decrease in microglial activation.