Background: Inflammation is one of the primary mechanisms behind the onset and advancement of renal disease in salt-sensitive hypertension (SS-HTN). One of the key mediators of inflammation is histamine, and there is a gap in knowledge regarding renal effects of histamine. We have shown that Dahl SS rats (a model of SS-HTN) exhibit elevated renal histamine levels. Further, we have reported that kidneys express all components of the histaminergic system and are capable of local histamine production. Highly abundant in the kidney, enzyme histamine N-methyltransferase (HNMT) is essential for the regulation of histamine levels. We hypothesized that the inhibition of HNMT-induced degradation of histamine would improve blood pressure and reduce renal tissue damage in the Dahl SS rat. Methods: At 8 weeks of age, male Dahl SS rats fed an NS diet (NS, 0.4% NaCl, Dyets) were implanted with telemeters and s.c. osmotic pumps filled with SKF91488 (HNMT inhibitor, n=7) and vehicle (veh, n=8) with infusion rate 0.04 mg/hr. Then, the animals were placed on a high salt (HS, 4% NaCl, Dyets AIN-75-based) diet for 21 days to induce SS-HTN, and telemetric measurements of BP were carried out. Metabolic cage studies were performed at the beginning and at the end of the protocol. At the endpoint, plasma and urinary electrolytes, biometric data were analyzed. Picrosirius red stain (PSR) and kidney injury molecule-1 (KIM-1) staining were employed for histological analysis to assess renal fibrosis and damage. OriginPro ( t-test,1-way or 2-way ANOVA ) was used for statistical analysis. Results: At the end of the HS diet, mean arterial pressure was lower in the SKF91488 group compared to vehicle (on day 21, the MAP was 132.6±11.8 vs 160.3±6.5 mmHg, respectively, p<0.05 ); heart rates were similar between the groups (367.5±15.1 vs 363.4±5.3 BPM respectively, day 21, p>0.05). We report a significant increase in the water consumption, urinary output, Na + and Cl - excretion in both vehicle and SKF91488 groups at end point compared to the start date (p<0.05). KIM-1 levels were lower after SKF91488 treatment (1.7±0.2 vs 0.8±0.1 a.u. in veh vs SKF groups, respectively, p<0.05). The analysis of PSR staining indicated similar protein cast formation in SKF91488 compared to the vehicle group (scoring 2.3±0.4 vs 1.9±0.5 a.u. out of 5). Conclusion: The chronic infusion of HNMT inhibitor results in a reduction of blood pressure and attenuation of renal damage compared to vehicle-infused animals. This study provides new perspectives on the role of the renal histaminergic system in mediating SS-HTN mechanisms. R01 HL148114, AHA 24TPA1283630, U54HL169191 This abstract was presented at the American Physiology Summit 2025 and is only available in HTML format. There is no downloadable file or PDF version. The Physiology editorial board was not involved in the peer review process.

The exocyst and Ift88 are necessary for primary ciliogenesis. Overexpression of Exoc5 (OE), a central exocyst component, resulted in longer cilia and enhanced injury recovery. Mitochondria are involved in acute kidney injury (AKI). To investigate cilia and mitochondria, basal respiration and mitochondrial maximal and spare respiratory capacity were measured in Exoc5 OE, Exoc5 knockdown (KD), Exoc5 ciliary targeting sequence mutant (CTS-mut), control Madin-Darby canine kidney (MDCK), Ift88 knockout (KO), and Ift88 rescue cells. In Exoc5 KD, Exoc5 CTS-mut, and Ift88 KO cells, these parameters were decreased. In Exoc5 OE and Ift88 rescue cells they were increased. Reactive oxygen species were higher in Exoc5 KD, Exoc5 CTS-mut, and Ift88 KO cells compared with Exoc5 OE, control, and Ift88 rescue cells. By electron microscopy, mitochondria appeared abnormal in Exoc5 KD, Exoc5 CTS-mut, and Ift88 KO cells. A metabolomics screen of control, Exoc5 KD, Exoc5 CTS-mut, Exoc5 OE, Ift88 KO, and Ift88 rescue cells showed a marked increase in tryptophan levels in Exoc5 CTS-mut (113-fold) and Exoc5 KD (58-fold) compared with control cells. A 21% increase was seen in Ift88 KO compared with rescue cells. In Exoc5 OE compared with control cells, tryptophan was decreased 59%. To determine the effects of ciliary loss on AKI, we generated proximal tubule-specific Exoc5 and Ift88 KO mice. These mice had loss of primary cilia, decreased mitochondrial ATP synthase, and increased tryptophan in proximal tubules with greater injury following ischemia-reperfusion. These data indicate that cilia-deficient renal tubule cells are primed for injury with mitochondrial defects in tryptophan metabolism.NEW & NOTEWORTHY Mitochondria are centrally involved in acute kidney injury (AKI). Here, we show that cilia-deficient renal tubule cells both in vitro in cell culture and in vivo in mice are primed for injury with mitochondrial defects and aberrant tryptophan metabolism. These data suggest therapeutic strategies such as enhancing ciliogenesis or improving mitochondrial function to protect patients at risk for AKI.

Atrial Natriuretic Peptide (ANP) plays an important role in blood pressure regulation. Low levels of ANP correlate with the development of salt-sensitive hypertension (SS-HTN). Our previous studies indicated that ANP deficiency exacerbated renal function decline in SS-HTN. In the heart and fat tissue, ANP was reported to affect lipid peroxidation and mitochondrial bioenergetics but the effects of ANP on mitochondrial function in the kidney are unexplored. We hypothesized that ANP deficiency in SS-HTN causes renal bioenergetic shift, leading to disruption of mitochondrial network and oxidative stress. To address the hypothesis, we placed Dahl SS wild-type (SSWT) and ANP knockout (SSNPPA-/-) rats on 4% NaCl high salt (HS) diet to induce HTN or maintained them on 0.4% NaCl normal salt (NS) diet and assessed mitochondrial bioenergetics and dynamics using spectrofluorimetry, Seahorse assay, electron paramagnetic resonance (EPR) spectroscopy, Western blotting, electron microscopy, PCR and cytokine assays. We report that under high salt conditions, associated with hypertension and renal damage, the SSNPPA-/- rats exhibit a decrease in mitochondrial membrane potential and elevation in mitochondrial ROS levels compared to SSWT. The redox shift is also evident by the presence of more pronounced medullar lipid peroxidation in the SSNPPA-/- strain. We also revealed fragmented, more damaged mitochondria in the SSNPPA-/- rats, accompanied by increased turnover and biogenesis. Overall, our data indicate that ANP deficiency causes disruptions in mitochondrial bioenergetics and dynamics which likely contributes to aggravation of the renal damage and hypertension in the Dahl SS rat; the major pathological effects are evident in the groups subjected to a combined salt and ANP deficiency-induced mitochondrial stress.

Background. Inflammation is one of the key mechanisms for the development of salt-sensitive hypertension (SS-HTN) and damage to the kidney. We have previously reported that the kidneys express components of the histaminergic system, and are capable of local production of histamine, a well-known inflammatory mediator. We showed that renal histamine level is increased in SS-HTN. Histamine content is tightly regulated by the enzyme histamine-N-methyltransferase, which has higher abundance in the kidney compared to other organs. We hypothesized that an increasing histamine level by limiting its breakdown will have beneficial effects on blood pressure (BP) and kidney damage in SS-HTN. Methods. Male Dahl SS rats at 9 weeks of age fed a NS diet (NS, 0.4% NaCl, Dyets) were implanted with telemeters and s.c. osmotic pumps filled with SKF91488 (N-methyltransferase inhibitor, prevents histamine breakdown, n=5) and vehicle (veh, n=5) with infusion rate 0.03875 mg/hr. Then, the animals were placed on a high salt (HS, 4% NaCl, Dyets AIN-75-based) diet for 21 days to induce SS-HTN. Telemetric measurements of BP were carried out. Metabolic cage studies were performed at the beginning and at the end of the protocol. At the endpoint, glomerular filtration rate (GFR) was measured; plasma and urinary electrolytes were analyzed. PSR and Masson Trichrome staining were employed for histological analysis. OriginPro ( t-test,1-way or 2-way ANOVA) were used for statistical analysis. Wire myography was performed for vascular relaxation analysis in renal or interlobular arteries from rats on NS and HS diets subjected to histamine (no SKF infusion). Results. At the end of the HS diet, MAP was lower in the SKF91488 infusion group compared to veh group (148.7±10.9 vs 162.9±10.8, 144.2±12.6 vs 166.1±11.1, 143.3±14.2 vs 171.1±10.5 mmHg on days 18-20 of HS diet, p<0.05). GFR was lower in the SKF91488 rats compared to veh group (0.44±0.05 vs 0.56±0.04 mL/min/100gBW, respectively, p=0.07). Electrolyte analysis showed trends towards higher urinary Na+ and Cl- excretion in the SKF91488 group ( p=0.089 and p=0.07, respectively). Urine flow, plasma electrolytes, kidney/body weight and water intake were similar. A significant amount of protein casts was found in the SKF91488 compared to vehicle group (8.31±0.76 vs 5.12±1.04%, p<0.05); pathology assessment revealed recruitment of immune cells into the kidney cortex in the SKF91488 group. Vascular relaxation dose responses to histamine were similar in HS rats vs NS rats (no SKF; p>0.05). Conclusion. We report that chronic administration of a histamine-N-methyltransferase inhibitor significantly reduced BP in the Dahl SS rats, whereas protein cast formation and recruitment of immune cells in the renal tissue was augmented. This study offers new insights into the role of the renal histaminergic system in the development of SS-HTN and renal damage and suggests potential differential effects of histamine in vasculature and renal epithelium. NIH R01HL148114 and AHA 23POST1020105 This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

There are numerous reports that male Dahl rats have greater increases in BP with a high-salt (HS) diet than females, yet women have been suggested to be more salt-sensitive than men. Since the typical Western diet consists of both HS and high-fat (HF), the goal of the present study was to test the hypothesis that HF loading and a combined HF/HS would abolish the protection observed in female Dahl rats vs. males. To test our hypothesis, 6 week old male and female Dahl rats were implanted with telemetry devices to continuously measure mean arterial BP; n=6. Rats were allowed to recover for one week followed by one week of baseline BP recording. Rats were then placed on a HF diet (60% kcal from lard; Bio-Serv) for 2 weeks followed by a HF/HS (4% salt; Bio-Serv) for an additional 2 weeks. Rats were placed in metabolic cages prior to starting HS and after 2 weeks to collect a 24 hour urine sample to measure urinary protein and albumin excretion. Glomerular filtration rate (GFR) was measured at the end of the study conscious freely moving animals using FITC-inulin elimination method. Baseline BP values were comparable between males (115±5 mmHg) and females (117±3 mmHg; p=0.67). BP increased during HF diet treatment, yet BP values remained similar in males and females (133±7 vs. 134±3 mmHg, respectively; p=0.93). Interestingly, females had a greater increase in BP on HF/HS vs. males that reached significance by the end of the study (146±7 vs. 160±6 mmHg, respectively; p=0.03). Despite this, urinary protein (185±23 vs. 94±19 mg/day; p=0.001) and albumin (142±22 vs. 56±13 mg/day; p=0.006) excretion were greater in males than in females. However, GFR was comparable between males and females (0.55 ± 0.03 vs. 0.45 ± 0.04 ml/min/100 g of body weight; p=0.07). In conclusion, female Dahl rats are not protected from HS diet when also challenged with HF, supporting our hypothesis that HF loading eliminates cardiovascular protection in females vs. males. However, males exhibited greater renal injury. This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Hypoxia occurs in situations of disbalance between metabolic needs and the supply of oxygen to organs and tissues of the body. In this regard, tissue hypoxia and ischemia are essential components of the pathogenesis of many diseases. One of the promising areas of research into the mechanisms of ischemia is attempting to weaken the negative effect of hypoxia and ischemia in the brain by using a variety of techniques that activate neuroprotective mechanisms. Here, we aimed to assess the dynamics of restoration of motor activity control in an experimental model of ischemic stroke in rats (cerebral ischemia, CI) after intranasal perineural implantation of mesenchymal stem cells into the receptive field of the olfactory nerve. It was found that the perineural administration of MSCs to rats in the acute period of cerebral ischemia was accompanied by clear signs of recovery of cognitive and motor functions within 1 and 3 days after the operation. On the seventh day after ischemia modeling, rats with the introduction of MSCs had no distinctive features in the control of motor activity compared to the period before the operation in the same rats. In the hippocampus of rats after modeling ischemia, a significant decrease in the content of NO by about 50% relative to the initial level is observed after 1 day. In the hippocampus of rats in which ischemia was modeled with simultaneous intranasal administration of MSC, a significant decrease in NO content by 39% relative to the initial level was also observed after 1 day. The content of NO increases slightly, but the difference in the level of NO relative to ischemic rats was not significant. The copper content in the hippocampus in the rats of these two groups did not change. There was a tendency to increase the efficiency of the antioxidant system 1 day after ischemia in both studied groups, and this effect was more pronounced with intranasal administration of MSC.

Introduction: Although nicotine’s harmful effects on renal function are established, the precise cellular mechanisms of smoking-related damage are understudied. Smoking-Related Glomerulopathy (SRG) is a renal disease phenotype associated with smoking. This condition histologically mimics diabetic nephropathy, however, SRG patients present with proteinuria and renal insufficiency without diabetes. Here we investigated the acute and chronic nicotine-related oxidative and nitrosative stress in glomerular podocytes. We hypothesized that nicotine may promote nitrosative stress in the kidney cells through the rapid production of peroxynitrite (ONOO - ) and a decline in nitric oxide (NO) bioavailability. Methods: To test the hypothesis, we used a conditionally immortalized human podocyte line and confocal imaging to detect the production of ONOO - (Hydroxyl Radical and Peroxynitrite Sensor; HPF), intracellular Ca 2+ (Fluo-8), and NO (DAF-FM). The presence of nicotinic acetylcholine receptors (nAChR) receptor subunits in podocytes was confirmed with immunocytochemistry and live imaging using commercially available pharmacology. NOS activity was analyzed in response to Ang II with the specific blockers for NOS1 (NΩ-Propyl-L-arginine hydrochloride) and NOS2 (L-NIL) subunits. One-way ANOVA (OriginPro) was used for statistical analysis. Results: Immunostaining indicated that human and rat kidneys express nicotinic acetylcholine receptors (nAChR). Notably, we detected specific expression of α7 nAChR in glomerular podocytes. In podocytes, acute nAChR activation promoted the mobilization of intracellular Ca 2+ controlled by intracellular store activation, and fast ONOO - transients. Multiple consecutive applications of nicotine resulted in repeated intracellular Ca 2+ and ONOO - transients. Nicotine-mediated ONOO - response was efficiently blocked in the presence of superoxide dismutase (SOD). The application of specific α7 or α4β2, α2β4, α4β4 and α3β4 nAChR agonists elicited Ca 2+ transients but did not reproduce the ONOO - response to nicotine, suggesting that nitrosative processes may occur independently from Ca 2+ influx or nAChR function. Chronic exposure to nicotine (12-hrs) resulted in a significant decline in podocytes’ NO bioavailability (p<0.05). While under normal conditions, NO is produced primarily by NOS1 (70%), and the rest is attributed to NOS2, chronic nicotine exposure led to an elevation of NOS2 (75%) and a decline in NOS1 activity (p<0.001). We observed similar NOS remodeling under hyperglycemic conditions, suggesting similar nitrosative processes in response to nicotine and high glucose. Conclusions: Nicotine promotes superoxide-stimulated nitrosative stress and peroxynitrite formation in podocytes leading to decline in NO bioavailability and pathological activation of NOS2. R01 NIDDK DK126720 (OP), DK129227 (OP), HL148114 (DVI), NIH/NCATS/SCTR UL1TR001450/SCTR 2214 (OP). This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Histamine is a nitrogenous compound crucial for the inflammatory response. The knowledge regarding the renal effects of histamine is very limited. We showed that renal epithelia exhibit expression of the components of the histaminergic system. Furthermore, we revealed that there was a shift in the histaminergic tone in salt-sensitive rats when they were challenged with a high-salt diet. These data support the notion that histamine plays a role in renal epithelial physiological and pathophysiological functions.

Introduction. In 2020, more than 670,000 deaths in the United States had hypertension as a primary or contributing cause. Females generally exhibit less susceptibility to hypertensive renal damage compared to males. It is established that renal mitochondrial dysfunction and associated oxidative stress contribute to hypertension development. Our previous studies have shown that healthy renal mitochondria in the absence of hypertension exhibit sex differences in antioxidant protein abundance, ROS production, and oxygen consumption rates. We hypothesized here that in hypertensive state mitochondrial metabolism in male and female kidneys differentially regulates tissue damage and ultimately hypertension in females. Methods. To address our hypothesis, we used 11-week-old male and female Sprague Dawley rats infused with Angiotensin II (400 ng/kg/min to induce hypertension) or vehicle for 3 weeks via a s.c. osmotic pump. Mitochondria were isolated from the renal cortex and medulla of these rats; mitochondrial membrane potential and H2O2 production were measured by spectrofluorimetry using TMRM and Amplex Red dyes. Two-way ANOVA was utilized for statistical analysis in OriginPro. Metabolic profiles of renal cortices and medullae were generated using UHPLC-HRMS, and metabolites were identified by retention time exact mass using MAVEN and MetaboAnalyst software. Results. Spectrofluorimetry showed decreased membrane potential in both male and female renal medulla in animals infused with Ang II compared to vehicle (p<0.001), while in cortex this difference was only recorded in females (p=0.03). Ang II-infusion reduced H2O2 production in the renal cortex and medulla of female rats (p<0.001) compared to sham groups, while this was not observed in males. In both Ang II and vehicle-infused groups, females maintained higher H2O2 production compared to males. UHPLC-HRMS identified 147 mitochondria-related metabolites in each group. Ang II infusion increased TCA cycle and purine metabolites as well as amino acids in the cortex of both sexes and in male medulla, whereas these changes were not revealed in the female medulla. Interestingly, females exhibited a decrease in amino acid metabolism in the renal medulla. In contrast to the male cortex, the female cortex is responding to oxidative stress induced by hypertension by altering arginine biosynthesis and metabolism. Arginine is a biosynthetic precursor to putrescine, which is involved in polyamine biosynthesis. Conclusions. Male and female kidneys employ divergent metabolic mitochondria-related pathways in hypertensive state. The male cortex may rely more on protein degradation to produce uric acid, an antioxidant, while the female cortex utilizes protein degradation and polyamine biosynthesis in oxidative stress management. R01HL148114 This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Electron paramagnetic resonance (EPR) spectroscopy was used to record the content of nitric oxide (NO) and copper in brain tissues (frontal lobes and hippocampus) and liver of healthy rats and rats after ischemia modeling. Ischemia was simulated by ligation of the carotid arteries, followed by taking 3 ml of blood from the common carotid artery. Signals from triple complexes (DETC) were recorded by EPR spectroscopy of complexes (DETC)2-Fe2+-NO and Cu(DETC)2. Based on direct measurements by EPR spectroscopy, it was shown that a day after the modeling of ischemia, NO production in the hippocampus decreases by an average of 30% and there is a tendency to decrease NO in the frontal lobes and liver. The copper content decreased by an average of 3 times in the frontal lobes and the hippocampus by an average of 20% a day after ischemia modeling, and a tendency to decrease was noted in the liver. Thus, brain hypoxia is accompanied not only by a decrease in NO production, but also by signs of weakening of the antioxidant system in the hippocampus and frontal lobes, which further worsens the functional state of the homeostasis system.

Electron paramagnetic resonance (EPR) spectroscopy was used to record the content of nitric oxide (NO) and copper in brain tissues (frontal lobes and hippocampus) and liver of healthy rats and rats after ischemia modeling. Ischemia was simulated by ligation of the carotid arteries, followed by taking 3 ml of blood from the common carotid artery. Signals from triple complexes (DETC) were recorded by EPR spectroscopy of complexes (DETC) 2 -Fe 2+ -(NO) and Cu(DETC) 2 . Based on direct measurements by EPR spectroscopy, it was shown that a day after the modeling of ischemia, NO production in the hippocampus decreases by an average of 30% and there is a tendency to decrease NO in the frontal lobes and liver. The copper content decreased by an average of 3 times in the frontal lobes and the hippocampus by an average of 20% a day after ischemia modeling, and a tendency to decrease was noted in the liver. Thus, brain hypoxia is accompanied not only by a decrease in NO production, but also by signs of weakening of the antioxidant system in the hippocampus and frontal lobes, which further worsens the functional state of the homeostasis system. Keywords: electron paramagnetic resonance, spin trap, nitric oxide, cerebral ischemia, frontal lobes, hippocampus.

A comparative experimental analysis of intensity of nitric oxide (NO) production and the copper content in the tissues of hippocampus of male Wistar rats after modeling of hemorrhagic stroke and brain injury was conducted using EPR spectroscopy. Modeling of hemorrhagic stroke was carried out by microinjection of 500 nl of autologous blood into the brain to a depth of 5.0 mm (hippocampus) on the left side. Brain injury was performed by removing a piece of nerve tissue from 5.0 mm depth on the left side of hippocampus. It was registered a significant decrease in the NO content in hippocampus by 36 ± 17% on the 3rd day after modeling of hemorrhagic stroke together with decrease by an average of 24 ± 14% of the copper content. There were no significant changes in the NO level in hippocampus found neither on the 3rd day nor on the 7th day after brain injury modeling. There was also no change in copper content. Thus, it was experimentally demonstrated that modeling of brain injury, in contrast to hypoxia induced by hemorrhagic stroke, was not accompanied with significant changes in NO production in hippocampus of rat.

Relevance. The problem of effective prevention and treatment of traumatic brain injuries (TBI) of various etiologies has not been resolved in all countries of the world. Primary brain damage from trauma initiates secondary damage to the nervous tissue. As a result, the interaction of brain neural networks is disrupted and the control of somatic and visceral functions of the body is weakened. The article is based on our own clinical observations and comparison of results with literature data and provides a discussion of the prospects for the use of cell technologies in the prevention of fatal disorders of vital functions control in traumatic brain injuries. Objective. To evaluate the effectiveness of intranasal perineural implantation of mesenchymal stem cells (MSCs) in the complex therapy of patients with TBI. Materials and methods. The technique intranasal perineural administration of MSCs was used in complex therapy of 15 patients with severe TBI. The patients were 19÷69 years old, 13 men and two women. A cell suspension was isolated from the adipose tissue of the patient's abdominal wall and centrifuged for 10 min at 1500 rpm. The cell pellet was washed in phosphate buffered saline and DMEM. Cells were cultured in plastic culture flasks in a humidified atmosphere with 5% CO2 content. The cell mass was trypsinized according to standard technique and resuspended in physiological saline on the day of implantation. Dynamics of culture growth, pluripotency, phenotyping of MSCs were monitored. MSCs were injected under general anesthesia into the submucosa of the nasal cavity 3-4 times with an interval of 3-7 days, depending on the growth rate of MSCs, in a single dose from 12.0×106 to 35.0×106 cells. Results. The use of allogeneic and predominantly autologous MSCs of adipose tissue in the complex treatment of patients with severe TBI by intranasal perineural delivery to the area of traumatic brain injury does not cause complications and is a safe technique. 8 patients with severe TBI showed from 4 to 7 points according to the Glasgow Outcome Scale Extended, with an average of 5.4±1.1 points after 6 months. The main result is that complex therapy, including intranasal implantation of MSCs in acute and subacute periods of severe TBI, contributes to the survival of patients and restoration of neurological – including cognitive – functions control. Conclusions. The effectiveness of intranasal perineural implantation of MSCs in the complex therapy of patients with TBI has been demonstrated. The mechanisms of the beneficial effects of perineural implantation of MSCs in patients with TBI require further research.

In order to simulate apoptotic processes in the central nervoussystem and retina, one eyeball was enucleated in each of the maleWistar rats (n = 10) aged 10–12weeks. Simultaneously, in one of the series of experiments (n = 5), 100 000 PKH67-labeled mesenchymalstem cells (MSCs) in 100 µL of buffer solution were injected parabulbaricallyon the side of an intact eye. In another series of experiments onrats (n = 5), immediately afterenucleation, on one side of the eye, 100 000 PKH67-labeled MSCsin 100 µL of buffer solution were injected intranasally into theupper part of the nasal cavity. Three days after enucleation, brainand retinal slices were prepared on a cryostat. In both series ofexperiments, the distribution of fluorescent MSCs was determinedin the mesencephalic quadruplet bodies, diencephalic lateral geniculatebody, and intact eye’s retina. It was inferred that intranasal perineuralimplantation of MSCs, due to its simplicity and efficiency, is apromising alternative method of MSC delivery into the retina ofpatients with signs of dystrophic and degenerative processes.

It was shown by electron paramagnetic resonance (EPR) spectroscopy that one day after bilateral occlusion of the common carotid arteries (to simulate ischemia) in Wistar rats (group 1, n = 15) under ketamine-xylazine-acepromazine anesthesia (55.6, 5.5, 1.1 mg/kg, respectively, intraperitoneally), the content of nitric monoxide (NO) in the olfactory bulbs decreased, and recovered after two days. A similar trend was observed in Wistar rats (group 2, n = 15), which, simultaneously with ischemia modeling, were implanted intranasally with mesenchymal stem cells (MSCs). An intact group of rats (group 3, n = 15) was kept in the usual conditions of the vivarium and did not undergo surgical interventions. The content of copper in the olfactory bulbs of rats (for assessing the activity of superoxide dismutases 1 and 3) after ischemia modeling (group 1) tended to increase and maintain the increase for two days of observation. Intranasal administration of MSCs during the ischemia modeling was accompanied by a more significant increase in copper content on the first day after brain ischemia modeling, but after two days, a tendency towards restoration of the initial copper level was noted.Intranasal administrating of MSC simultaneously with ischemia modeling (group 2) was accompanied by a more rapid recovery of orientational-motor activity in experimental animals compared to rats in which only modeling of ischemic stroke was performed (group 1).

Results of analysis of nitric oxide and copper content in rat liver and hippocampus after brain ischemia modeling are provided. The studies are carried out using the electron paramagnetic resonance spectroscopy method with spin traps. It was shown that, the day after brain ischemia modeling, nitric oxide content in hippocampus decreases on average by 50% and a tendency toward its decrease was observed in liver tissues. Two days after brain ischemia modeling, nitric oxide content in the brain recovered and a significant increase by 46% against control indices was observed in the liver. On the second day of the postischemic period, the copper content, which is associated with superoxide dismutase content, increased on average by 2.5-fold in the liver. No significant changes in copper content was found in the hippocampus.

The problem of treatment of oncologic patients is not resolved. Our experiments focused on the ability of autologous mesenchymal stem cells (MSCs) to inhibit glioma cell growth in vitro and in vivo. 28 thousand cells of C6 glioma in 20 μl of culture medium were stereotactically introduced into occipital region of brain (Oc2L) from the right side after craniotomy in experiments on 30 male Wistar rats weighing 250–270 g under ketamine‐xylazine‐acepromazine (55.6, 5.5, and 1.1 mg/kg, respectively) anesthesia. 20 rats received 400 thousand MSCs labeled with CD90s and FITC in 50 μl of culture medium under nasal mucosa during the operation. Brain slices 8 μm in thickness were prepared after decapitation of one rat from each group in one day after surgery. MSCs distribution was visualized using Zeiss AxioVert 200M fluorescence microscope with Zeiss AxioCam HRm CCD camera. Fluorescent MSCs were revealed only in damaged Oc2L area of the rat with previously injected MSCs. 10 rats from the same group (with preimplanted MSCs) received 400 thousand MSCs in 50 μl of culture medium via weekly spray injections into both nasal cavities. Rats with implanted glioma and without MSCs died in 15.0±4.3 days. Rats with implanted glioma and single intranasal MSCs injection died in 21.1±5.2 days. 8 of 10 rats that received both intranasal MSCs injection and subsequent weekly spray MSCs injections died in 33.6±3.4 days. Two rats are still alive (108 days from the date of operation). Morphological examination of brain slices of rat from the last group showed signs of apoptosis of tumor cells. It was assumed that MSCs trigger apoptosis in C6 glioma cells via paracrine way by excretion of exosomes and cytokines.

Pathological processes in cardiovascular system and liver are fatal for citizens of all over the world. New preventive and therapeutic approaches are needed in this field of medicine. Using of cell technologies for the treatment of patients with heart attack and/or cirrhosis is the one. Systemic or intraorganic stem cells (SCs) administration has shown poor therapy results. New technology was substantiated in our experiments on 27 male Wistar rats weighing 240–270 g. Vagus was cut off in 9 animals after cervical midsection on the left side under ketamine‐xylazine‐acepromazine (55.6, 5.5, and 1.1 mg/kg, respectively) anesthesia (I) in order to initiate destructive processes in heart. 9 rats were subjected to laparotomy followed by 2×2 mm injury of liver right lobe (II). Both laparotomy and cervical midsection but without liver and vagus damage was performed in the last 9 rats. All the rats received 10 thousands SCs in 10 μl culture medium into perineural space of vagus on the right side of the neck during anesthesia. SCs were marked with monoclonal antibodies to CD90 and PKH67 Green Fluorescent Cell Linker. Two animals from each group were decapitated in one day, one week and two weeks and slices from heart and liver 8 μm in thickness were prepared. SCs distribution was visualized using Zeiss AxioVert 200M fluorescence microscope with Zeiss AxioCam HRm CCD camera. Fluorescent cells were revealed in heart and liver areas in groups I and II, respectively. Zero fluorescent cells have been found in heart and liver of group III. Therefore, perineural way of SCs administration is an alternate path of SCs delivery to damaged inner organs only.