Rat Adipose Tissue Research Articles

High-Intensity Interval Training Does Not Change Vaspin and Omentin and Does Not Reduce Visceral Adipose Tissue in Obese Rats.

This study aimed to determine the expression of omentin and vaspin, inflammatory markers, body composition, and lipid profile in diet-induced obese rats and high-intensity interval training (HIIT). Forty Wistar rats were divided into four groups: untrained normal diet, trained normal diet (T-ND), untrained high-fat diet (Unt-HFD), and trained high-fat diet (T-HFD). For the animals of the Unt-HFD and T-HFD groups, a high-fat diet was offered for 4 weeks. After that, all the animals in the T-ND and T-HFD groups were submitted to HITT, three times per week, for 10 weeks (2 weeks of adaptation and 8 weeks of HIIT). Muscle (gastrocnemius), liver, epididymal adipose tissue, retroperitoneal adipose tissue, visceral adipose tissue (VAT), and serum were collected to analyze TNF-α, IL-6, PCR, IL-8, IL-10, IL-4, vaspin, and omentin. A body composition analysis was performed before adaptation to HIIT protocol and after the last exercise session using dual-energy X-ray absorptiometry. Omentin and vaspin in the VAT were quantified using Western blotting. The results showed that, when fed a high-fat diet, the animals obtained significant gains in body fat and elevated serum concentrations of vaspin and blood triglycerides. The HIIT was able to minimize body fat gain but did not reduce visceral fat despite the increase in maximum exercise capacity. Moreover, there was a reduction in the serum levels of adiponectin, IL-6, and IL-10. Finally, we concluded that, although the training protocol was able to slow down the weight gain of the animals, there was no reduction in visceral fat or an improvement in the inflammatory profile, including no changes in omentin and vaspin.

Oestrogen replacement fails to fully revert ovariectomy-induced changes in adipose tissue monoglycerides, diglycerides and cholesteryl esters of rats fed a lard-enriched diet

Menopause may be accompanied by abdominal obesity and inflammation, conditions accentuated by high-fat intake, especially of saturated fat (SFA)-rich diets. We investigated the consequences of high-SFA intake on the fatty acid (FA) profile of monoglycerides, diglycerides and cholesteryl esters from retroperitoneal white adipose tissue (RET) of rats with ovariectomy-induced menopause, and the effect of oestradiol replacement. Wistar rats were either ovariectomized (Ovx) or sham operated (Sham) and fed either standard chow (C) or lard-enriched diet (L) for 12 weeks. Half of the Ovx rats received 17β-oestradiol replacement (Ovx + E2). Body weight and food intake were measured weekly. RET neutral lipids were chromatographically separated and FAs analysed by gas chromatography. Ovariectomy alone increased body weight, feed efficiency, RET mass, leptin and insulin levels, leptin/adiponectin ratio, HOMA-IR and HOMA-β indexes. OvxC + E2 showed attenuation in nearly all blood markers. HOMA-β index was restored in OvxL + E2. OvxC showed significantly disturbed SFA and polyunsaturated FA (PUFA) profile in RET cholesteryl esters (CE). OvxC also showed increased monounsaturated FA (MUFA) in the monoglyceride diglyceride (Mono–Di) fraction. Similar changes were not observed in OvxL, although increased SFA and decreased PUFA was observed in Mono–Di. Overall, HRT was only partially able to revert changes induced by ovariectomy. There appears to be increased mobilization of essential FA in Ovx via CE, which is a dynamic lipid species. The same results were not found in Mono–Di, which are more inert. HRT may be helpful to preserve FA profile in visceral fat, but possibly not wholly sufficient in reverting the metabolic effects induced by menopause.

Undernutrition induces major alterations in the lipid droplets of white and brown adipose tissues in wistar rats

Several studies have shown a relationship between the distribution of fat mass around the organism, metabolic disorders, and an increased risk of morbidity and mortality. It has been demonstrated that in obese animals there is a big rise in the white fat deposits due to hyperplasia and hypertrophy of the adipocytes. Studies related to weight and health have been more popular regarding obesity rather than extreme caquexia or calorico-proteic deficiencies, but these states are interesting from the point of view of the preferential atrophy of certain organs that may help us in the understanding of undernourishment. Moreover, the discovery of beige adipose tissue has instigated thoughts around the roles played by the different cells in the adipose tissue as well as its adaptability in pathological states.In our study we carried out morphometric, morphological, and quantitative measurements of the adipose tissue in an animal model based on a 40−50% diet restriction in comparison to control animals.We have found a decrease in the size of white adipocytes together with a variation in the lipid droplet size of brown adipocytes in undernourished animals, what may be considered as possible transformations between the types of adipose tissues, and that could be caused by an adaptive phenomenon to the undernourished state.

Lactobacillus fermentum MCC2759 and MCC2760 Alleviate Inflammation and Intestinal Function in High-Fat Diet-Fed and Streptozotocin-Induced Diabetic Rats

The growing incidence of type 2 diabetes and obesity has become a worldwide crisis with increased socio-economic burden. Changes in lifestyle and food habits resulting in dysbiosis of the gut microbiota and low-grade inflammation are linked to the rising incidence. The aim of this study was to investigate the effects of potential probiotic Lactobacillus fermentum MCC2759 and MCC2760 on intestinal markers of inflammation using a high-fat diet (HFD)-fed model and a streptozotocin (STZ)-induced diabetic model. Lact. fermentum administration showed improved oral glucose tolerance compared with the model controls of HFD (AUC 1518) and STZ (628.8). Plasma insulin levels improved in the Lact. fermentum treated groups of HFD + MCC2759 (129 ± 4.24 pmol/L) and HFD + MCC2760 (151.5 ± 9.19 pmol/L) in HFD study, while in STZ diabetic study, the insulin levels were normalized with Lact. fermentum administration, for D + MCC2759 (120.5 ± 7.77) and D + MCC2760 (138 ± 5.65 pmol/L) groups. The results showed reduction in inflammatory tone in liver, muscle, and adipose tissues of rats in both models with stimulation of anti-inflammatory IL-10 by real-time quantitative polymerase chain reaction. Additionally, the potential probiotic cultures also displayed normalization of markers related to intestinal barrier integrity (ZO-1), TLR-4 receptor, and insulin sensitivity (GLUT-4, GLP-1, adiponectin). Thus, the results suggest that Lact. fermentum could act as potential probiotic for lifestyle-related disorders such as obesity, diabetes, and metabolic syndrome as both prophylactic and adjunct therapies.

Variations in the mRNA expression level of UDP-GlcNAc epimerase/ManNAc kinase and neuraminidase 1 genes in organs of type 2 diabetic animals.

Sialic acid and its associated metabolic enzymes have emerged as important components of the pathophysiology of type 2 diabetes (T2D). There is an elevation in the serum concentration of sialic acid in humans and animals with T2D. The present study investigated the modulation of mRNA expression level of UDP-N-acetylglucosamine-2-epimerase/N-acetylmannosamine kinase (GNE) and neuraminidase 1 (NEU1) genes in some organs of type 2 diabetic rats. T2D was induced using fructose-streptozotocin model and eight weeks after the induction of diabetes, sialic acid was assayed in the blood and organs (adipose tissue, brain, colon, kidney, liver, pancreas, skeletal muscle and spleen) followed by quantification of mRNA expression level of GNE and NEU1 genes by qPCR. The results showed a significant (P < 0.05) increase in sialic acid level in the serum and all the afore-mentioned organs investigated except in the adipose tissue and skeletal muscle of the diabetic rats compared the normal control. The expression GNE gene was only increased in the pancreas (1.8-fold) of the diabetic rats while there was a decrease in the expression of the gene in the colon. In contrast, the expression of NEU1 gene was increased in the spleen (3.5-fold), brain (2.2-fold), liver (1.9-fold), colon (1.5-fold) and kidney of the diabetic rats. It was concluded that the elevated level of sialic acid in the organs of diabetic rats, except the pancreas, might not be due to increased endogenous synthesis of sialic acid.

Effects of curcumin administration on Nesfatin-1 levels in blood, brain and fat tissues of diabetic rats.

We evaluated the efficacy of curcumin administration on blood glucose levels and its relationship with nesfatin-1 levels in blood brain and adipose tissue of streptozotocin-induced diabetic rats. A total of 28 male rats were divided into four groups: control group, type 2 diabetes mellitus (DM) group, control plus curcumin group and type 2DM plus curcumin group. After fifteen days, blood samples were collected from sacrificed rats. Nesftain-1 levels were analysed from blood, brain, and fat tissues of rats in all groups. Nesfatin-1 level was found to be significantly lower in blood, brain and fat tissues of type 2 DM rats compared to the control group. A significant decrease in fasting blood glucose levels was observed in the curcumin administration group compared to type 2 DM group. Improvement of fasting blood glucose level was accompanied by improvement of nesfatin-1 levels in blood, brain, and fat tissues. As expected, curcumin administration caused significant improvement in fasting blood glucose levels. However, for the first time, we found marked improvements in nesfatin-1 levels in blood, brain, and fat tissues of type 2 DM rats. Thus, considering the crucial role of nesfatin-1 in regulation of glucose metabolism, it is logical to expect an interactive relationship between curcumin and nesfatin-1.

Lutein attenuates excessive lipid accumulation in differentiated 3T3-L1 cells and abdominal adipose tissue of rats by the SIRT1-mediated pathway.

Obesity is now a worldwide disease and is mainly attributable to increased body fat deposition. In a growing number of epidemiological studies, lutein has been revealed to have different degrees of anti-obesity properties, but the potential underlying mechanisms that have been reported are limited. Therefore, we aimed to clarify the protective effects of lutein against excessive lipid accumulation, and we explored the role of SIRT1 and SIRT1-mediated pathways both in abdominal adipose tissue and mature 3T3-L1 cells during lutein administration. In our design, male Sprague-Dawley rats were fed either control or high-fat diets with or without 25 mg/kg·bw/day lutein for 5 weeks. Additionally, differentiated 3T3-L1 cells were incubated with 40 μM lutein or 10 μM Ex527 for 24 h. Lutein supplementation decreased the body weight, abdominal fat index ratio, frequency and mean area of larger adipocytes in HE staining induced by the high-fat diet and then activated the expression of SIRT1 and thus upregulated FoxO1, ATGL, and HSL expression and downregulated SREBP-1, FAS, and ACC expression both in abdominal adipose tissue and differentiated 3T3-L1 cells. However, coincubation with Ex527 and lutein suppressed the activation of SIRT1 and reversed the expression of FoxO1, ATGL, HSL, SREBP-1, FAS, and ACC in comparison to those in the Lut group. Overall, we suggest that the effects of lutein on attenuating excessive lipid accumulation are dependent on the SIRT1-mediated pathway in vivo and in vitro, which indicates that lutein administration may be a potential strategy for preventing excessive lipid accumulation and obesity.

Effects and Mechanisms of Chitosan and ChitosanOligosaccharide on Hepatic Lipogenesis and Lipid Peroxidation, Adipose Lipolysis, and Intestinal Lipid Absorption in Rats with High-Fat Diet-Induced Obesity.

Chitosan and its derivative, chitosan oligosaccharide (CO), possess hypolipidemic and anti-obesity effects. However, it is still unclear if the mechanisms are different or similar between chitosan and CO. This study was designed to investigate and compare the effects of CO and high-molecular-weight chitosan (HC) on liver lipogenesis and lipid peroxidation, adipose lipolysis, and intestinal lipid absorption in high-fat (HF) diet-fed rats for 12 weeks. Rats were divided into four groups: normal control diet (NC), HF diet, HF diet+5% HC, and HF diet+5% CO. Both HC and CO supplementation could reduce liver lipid biosynthesis, but HC had a better effect than CO on improving liver lipid accumulation in HF diet-fed rats. The increased levels of triglyceride decreased lipolysis rate, and increased lipoprotein lipase activity in the perirenal adipose tissue of HF diet-fed rats could be significantly reversed by both HC and CO supplementation. HC, but not CO, supplementation promoted liver antioxidant enzymes glutathione peroxidase and superoxide dismutase activities and reduced liver lipid peroxidation. In the intestines, CO, but not HC, supplementation reduced lipid absorption by reducing the expression of fabp2 and fatp4 mRNA. These results suggest that HC and CO have different mechanisms for improving lipid metabolism in HF diet-fed rats.

The Effect of Swimming Training and Cinnamon Consumption on the Gene Expression of GLUT4 and Insulin Receptor in the Brown Adipose Tissue of Diabetic Rats

Background: The effect of training on the gene expression of GLUT4 and insulin receptor (IR) has been investigated in some studies, but the simultaneous effect of swimming training along with cinnamon consumption is unknown. Objective: This study aimed to examine the effect of six weeks of swimming training with cinnamon consumption on the gene expression of GLUT4 and IR in the brown adipose tissue of diabetic rats. Methods: In this experimental study, 28 diabetic rats were randomly divided into four groups of seven animals, including 1- control (C), 2- cinnamon (Ci), 3- swimming (S), and 4- swimming plus cinnamon (S + Ci). Rats in groups 3 and 4 trained for six weeks and five sessions. Groups 2 and 4 received 200 mg/kg/day. Data were analyzed using one-way analysis of variance and Tukey’s post hoc test at the level of P ≤ 0.05. Results: The gene expression of GLUT4 and IR in the S, Ci, and S + Ci groups was significantly (P = 0.001) higher than the control group. Also, the gene expression of GLUT4 and IR in the S group was significantly (P = 0.001) higher than the Ci and S + Ci groups. Conclusion: Swimming training and cinnamon consumption and their simultaneous implementation had a significant effect on increasing the gene expression of GLUT4 and IR in the brown adipose tissue of diabetic rats. On the other hand, swimming training alone had a greater effect than cinnamon consumption and swimming plus cinnamon consumption.

Nutmeg extract potentially alters characteristics of white adipose tissue in rats.

BackgroundBrowning of white adipose tissue (WAT) is a promising approach to obesity treatment. During browning, WAT transforms into beige adipose tissue through stimulation of the peroxisome proliferator activated receptor γ (PPARγ). Nutmeg, one of the Indonesian herbs, reportedly has dual roles as a PPARα/γ partial agonist. Even though nutmeg has been traditionally used in body weight reduction, there is limited information regarding the potential role of nutmeg in browning of WAT.ObjectivesIn this study, we explored the effect of nutmeg seed extract (NuSE) as a potential inductor of WAT browning.MethodsTwelve male Wistar rats, 5–6 weeks old, were divided into control and nutmeg groups. The rats in nutmeg group were given NuSE for 12 weeks by oral gavage. After 12 weeks, the rat's inguinal WAT and brown adipose tissue (BAT) were collected, weighed and stored at − 80°C until use.ResultsWe observed that even though NuSE did not reduce the final body weight, it significantly reduced body weight gain. NuSE also increased protein levels of peroxisome proliferator activated receptor γ coactivator 1α (PGC‐1α) and uncoupling protein 3 (UCP3) significantly and tended to increase UCP2 and UCP1 levels. Furthermore, NuSE induced macroscopic and microscopic morphological changes of inguinal WAT, marked by significantly increased adipocyte numbers and decreased adipocyte size.ConclusionsEven though NuSE did not increase UCP1 significantly, it potentially alters inguinal WAT characteristics and leads to browning through PGC‐1α and UCP3 induction. However, UCP3’s specific mechanism in WAT browning remains unclear. Our findings could contribute to obesity treatment in the future.

Tart cherry (Prunus cerasus L.) dietary supplement modulates visceral adipose tissue CB1 mRNA levels along with other adipogenesis-related genes in rat models of diet-induced obesity.

There is increasing evidence for the involvement of dietary bioactive compounds in the cross-talk modulation of endocannabinoid system and some of the key regulators of transcriptional control for adipogenesis. We aimed to characterize the expression of cannabinoid CB1/CB2 receptors and fatty acid amide hydrolase (FAAH) along with selected adipogenesis-related genes (PPARγ, SREBP-1c and PREF-1), adipocyte-secreted factors (leptin and adiponectin), mitochondrial bioenergetic modulators (PGC-1A and UCP-2), and transient receptor potential vanilloid subtype 1 (TRPV1) and 2 (TRPV2) channels in visceral adipose tissue of rats fed with a high-fat diet (HFD) containing either tart cherry seeds alone or tart cherry seeds and juice for 17weeks. The visceral adipose tissue was weighed and checked the expression of different markers by qRT-PCR, Western blot and immunohistochemistry. Tart cherry supplements were able to downregulate the HFD-induced mRNA expression of CB1 receptor, SREBP-1c, PPARγ, leptin, TRPV1 and TRPV2 resulting in potential anti-adipogenic effects. The present study points out that the intake of bioactive constituents of tart cherry may attenuate the effect of adipogenesis by acting directly on the adipose tissue and modulating the interplay between CB1, PPARγ and TRPV channel gene transcription.

Preliminary study on effect of Gegen Qinlian Decoction on enzyme activity, gene expression and methylation level of FASN in adipose tissue of rats with insulin resistance

To investigate the effect of Gegen Qinlian Decoction(GQD) on enzyme activity, gene expression and methylation level of fatty acid synthase(FASN) in adipose tissue from rats with insulin resistance induced by high-fat diet. The 60% fat-powered high-fat diet was continuously given to male SD rats to induce the insulin resistance model. Then, they were divided into five groups randomly and administrated by gavage every day for 16 weeks with following drugs respectively: 10 mL·kg~(-1)water for control group(C) and insulin resistance model control group(IR), 1.65 g·kg~(-1)GQD per day for low-dose group(GQDL), 4.95 g·kg~(-1)GQD per day for medium-dose group(GQDM), 14.85 g·kg~(-1)GQD per day for high-dose group(GQDH), and 5 mg·kg~(-1) rosiglitazone per day for rosiglitazone group(RGN). Epididymal adipose tissue was taken to determine enzyme activity of FASN by colorimetric method, mRNA expression level of Fasn by quantitative Real-time PCR(Q-PCR) and CpGs methylation level between +313 and +582 by bisulfite sequencing PCR(BSP). These results showed that Fasn expression was significantly lowered in IR model rats compared with the control rats(P&lt;0.01). Enzymatic activity and CpGs methylation level of Fasn in IR group showed downward trends. Low and medium-dose GQD can increase enzyme activity of FASN(P&lt;0.05). Moreover, low-dose GQD increased the total CpGs methylation level of Fasn fragment between +313 and +582 in insulin resistance rats(P&lt;0.05). For GQDM group, the methylation frequency of CpGs at positions +506 and +508(P&lt;0.01) as well as the methylation frequency of CpGs on the binding sites of transcription factorzinc finger protein 161(P&lt;0.05) were significantly increased. The methylation frequency of CpG at +442 position was positively correlated with Fasn expression(P&lt;0.01, r=0.735), and methylation frequencies of CpGs at +345 and +366 positions were positively associated to enzyme activity of FASN respectively(P&lt;0.05, r=0.479; P&lt;0.01, r=0.640). In conclusion, GQD can reverse enzyme activity of FASN and methylation level of Fasn in adipose tissue of insulin resistant rats, and CpG sites at positions +506 and +508 may be the targets of GQD. The methylation level of CpGs at + 345 and + 366 sites were possibly related to FASN activity, while methylation of CpG at + 442 site may be closely correlated with mRNA level of Fasn. In addition, GQD did not significantly change mRNA expression level of Fasn, but effectively reversed enzymatic activity, suggesting that GQD may regulate the post transcriptional expression of Fasn.

АНАЛІЗ ЯКІСНОГО СКЛАДУ МАТРИКСНИХ МЕТАЛОПРОТЕЇНАЗ У ПАТОГЕНЕЗІ ОЖИРІННЯ, ЩО ІНДУКОВАНЕ ВИСОКОКАЛОРІЙНОЮ ДІЄТОЮ У ЩУРІВ

The problem of obesity in modern life is becoming more actual and is a serious social risk for human life. This problem is common despite social and professional affiliation, location, age and gender. Obesity is associated with an increase growth of adipose tissue. Today obesity isn't studied only as an excessive accumulation of fat in the body, but as a chronic multivariate disease associated with a number of serious metabolic, oncological, cardiological and other complications. Although a lot of research studies regulation and intracellular processes of adipogenesis, however the information about molecular mechanisms of remodeling of the extracellular matrix during an increase fat mass is limited. Matrix metalloproteinases (MMPs) can increase the plasticity of the matrix, thereby providing adipose tissue remodeling and / or adipocyte hypertrophy, play an additional role in the growth of adipose tissue associated with obesity, supporting the differentiation of adipose tissue progenitor cells. Our research is aimed at characterization of enzymatic activitiesof MMPs in adipose tissue of the rats with obesity. We found that active enzymes with a molecular weight &gt; 100 kDa were found in the adipose tissue of rats for the development of obesity, which may result from the formation of homodimers in these pathological conditions.Also activities of the MMP-2, -9 increase during obesity. Future studies of enzymatic activity of MMPs of rat's adipocytes tissues will improve understanding the biochemical processes under the conditions of this pathology and development of new approaches to diagnosis and treatment of obesity principles.

Histological Changes in Adipose Tissue: An Alarm When Methamphetamine Is Targeted for Weight Loss Purposes

BACKGROUNDMethamphetamine (METH) may be administered for weight loss purposes and to understand the METH side-effects more in details, this study aimed at determining the effect of METH on changes in adipose tissue in experimental rats.METHODSForty five male Wistar rats were randomly allocated to three equal groups. Group 1 was experimental receiving METH [0.4 mg/kg, subcutaneously (S/C), 0.6 mL/rat] for 3 weeks, group 2 was the sham group receiving normal saline (0.6 mL/rat, S/C) and the 3rd group was the control receiving distilled water, identically. The elevated plus maze test was used to confirm cognitive impairment and distraction as anxiety and to verify addiction to METH by assessing the percent time spent in open arm (OAT), the percent time spent in closed arm (CAT), the percent time spent in central parts and head dipping over the side of the maze. Adipose tissue was assessed histologically 7, 14 and 21-days after interventions. RESULTSA significant increase in anxiety level, and histologically inflammation, degeneration and necrosis in adipose tissue were visible after METH use. CONCLUSIONMETH use resulted in a significant inflammation and necrosis in adipose tissue denoting to the dangers of METH use, when recreationally targeted for weight loss purposes.

The Effects of Testosterone Therapy Combined with Swimming Exercise on Adipose Tissue and Biochemical Parameters in Male Obese Wistar Rats.

Exercise and anabolic steroids are anticipated to promote fat mass reduction and so to decrease the number of comorbidities related to excessive weight. The aim of this study was to verify the influence of aerobic exercise and the use of steroids on the accumulation of adipose tissue and on the biochemical limitations of Wistar rats nourished by a hypercaloric diet. Forty, young male Wistar rats were split into four groups: obese control (n=10), obese under treatment (n=10), obese under aerobic exercise (n=10) and obese under aerobic exercise and treatment (n=10). All animals were fed with a hypercaloric diet and animals under treatment received intramuscular testosterone. Body (weight and visceral fat) and blood (lipidogram, glucose, and liver enzymes) parameters were assessed. The group treated with aerobic exercise and testosterone revealed a reduction in body weight and visceral, perirenal, retroperitoneal and epididymal fats, accompanied by the blood levels of glucose, lactate, LDL-cholesterol, HDL-cholesterol, and lactate dehydrogenase; following high-intensity physical activity. The results support the theory that the combination of steroids and physical activity reduces the side-effects of androgenic-anabolic hormones and conveys benefits to some constraints.

ОСОБЛИВОСТІ ЗАПАЛЕННЯ БУРОЇ ЖИРОВОЇ ТКАНИНИ У ЩУРІВ З РОЗВИТКОМ ІНДУКОВАНОГО ДІЄТОЮ ОЖИРІННЯ ПІСЛЯ РІЗНИХ РЕЖИМІВ УВЕДЕННЯ МЕЛАТОНІНУ

One of the prominent obesity-related changes is the development of systemic low-grade proinflammatory state. Brown adipose tissue (BAT) may serve as a potential target for activation by melatonin to facilitate heat production and simultaneously stimulate lipolysis during obesity development. At the same time, melatonin is known to have immunomodulatory properties, which are performed via endocrine and paracrine signal pathways in variety cell types (including brown adipocytes)and change significantly during the day. Therefore, it can be used in a wide range of doses and at different times of the day (chronotherapeutic approach). Thus, the main goal of our research was to analyze the inflammation state of brown adipose tissue of rats during high-calorie diet induced-obesity development after different daily melatonin application in different regimes. Melatonin was administered by gavage for 7 weeks in dose 30 mg/kg 1 h before lights-off (HCD ZT11, M ZT11, evening), or 1 h after lights-on (HCD ZT01, M ZT01, morning). Tissue collagen content and leukocyte infiltration levels in BAT, detected by Van Gieson trichrome staining, were used as markers for the assessment of BAT inflammation state BAT. Propagation of obesity resulted in the increase of BATfibrosis level (the relative area occupied by collagen fibers) and tissue leukocyte infiltration in comparison to control rats. BAT fibrosis level after melatonin administrations to obese rats of HCD ZT01 and HCD ZT11 groups decreased to control values. Similar effects were observedinBAT tissue leukocyte infiltration after both regimes (HCD ZT01 and HCD ZT11 groups) of melatonin intake: this parameter decreased significantly, comparing to obese rats, but was still elevated, comparing to controls. At the same time, melatonin treatmentin morning or evening regimes did not have any impact on BAT fibrosis propagation and leukocyte infiltration in animals that consumed standard diet (M ZT01 and M ZT11 groups). To sum up, we suggest corrective properties of melatonin in context of chronic low-grade inflammation in obese rats BAT and suppose its wide potential for the therapeutic use combined with virtually absent side effects on BAT histophysiology of non-obese rats.

OSTEOPLASTIC PROPERTIES OF MULTIPOTENT MESENCHYMAL STROMAL CELLS OF ADIPOSE TISSUE

The aim: Determining the ability of samples based on MMSC – AT differentiating in the osteogenic direction. Materials and methods: The study was conducting at Bukovinian State Medical University, Chernivtsi, Ukraine. Adipose tissue samples were obtaining from the neck of 60 experimental animals (white Wistar rats). Multipotent mesenchymal stromal cells of adipose tissue were obtained by grinding adipose tissue of rats in 0.1% collagenase 1A . Alkaline phosphatase activity was assessing by using the Alkaline Phosphatase Detection Kit (Sigma, USA) according to the manufacturer’s protocol. Osteopontin gene expression was determining by immunocytochemical method. To determine the mRNA used the PCR method, which is associated with reverse transcription (RT-PCR) in the area of quantification of gene expression to the marker BGP. Results: On the 21st day of observations, the expression of mRNA encoding the BGP gene decreased in samples № 1 and № 3 to 35,800 ± 420.0 copies and to 35,000 ± 400.0 copies, p1&lt;0.01, p&gt;0.05. Also was observing growth of copies of the BGP gene in samples № 2 and № 4 in 2.1, р&lt;0.01 and 2.2 times, р-р2&lt;0.05, relative to the data in sample № 1. Conclusions: Comparative study of osteoplastic properties samples MMSC-AT showed that a larger number of cells differentiate into the osteoblasts in samples containing MMSC-AT + PRP (№ 2) and MMSC-AT + PRP + «Kolapan» (№ 4). This has been proven higher alkaline phosphatase activity, higher levels osteopontin expression, and higher levels BGP gene expression.

Melatonin reverses the loss of the anticontractile effect of perivascular adipose tissue in obese rats.

Perivascular adipose tissue (PVAT) undergoes functional changes in obesity. Increased oxidative stress is a central mechanism whereby obesity induces loss of the anticontractile effect of PVAT. Melatonin is an antioxidant that displays vasoprotective action in cardiovascular disease. Here, we sought to investigate whether melatonin would restore the anticontractile effect of periaortic PVAT in obesity. Male Wistar Hannover rats were treated for 10weeks with a high-calorie diet. Melatonin (5mg/kg/d, p.o., gavage) was administered for 2weeks. Functional findings showed that obesity-induced loss of the anticontractile effect of PVAT and treatment with melatonin reversed this response. Tiron [a scavenger of superoxide anion (O2 - )] restored the anticontractile effect of PVAT in aortas from obese rats, suggesting a role for reactive oxygen species (ROS) in such response. Decreased superoxide dismutase (SOD) activity and augmented levels of ROS were detected in periaortic PVAT from obese rats. These responses were accompanied by decreased levels of nitric oxide (NO) in PVAT. Treatment with melatonin restored SOD activity, decreased ROS levels, and increased NO bioavailability in PVAT from obese rats. Here, we first reported the beneficial effects of melatonin in periaortic PVAT in obesity. Melatonin reversed the adverse effects of obesity in PVAT that included overproduction of ROS, reduced SOD activity, and decreased bioavailability of NO. Therefore, PVAT may constitute an important target for the treatment of obesity-induced vascular dysfunction and melatonin emerges as a potential tool in the management of the vascular complications induced by obesity.

Improvement of Hypertriglyceridemia by Roasted Nelumbinis folium in High Fat/High Cholesterol Diet Rat Model.

Hypertriglyceridemia is a condition characterized by high triglyceride levels and is a major risk factor for the development of cardiovascular diseases. The present study was designed to investigate the inhibitory effect of roasted Nelumbinis folium (RN), which is a medicinal substance produced by heating lotus leaves, on lipid metabolism in high fat/cholesterol (HFC) diet-induced hypertriglyceridemia. Except for those in the control group, Sprague–Dawley rats were fed an HFC diet for four weeks to induce hypertriglyceridemia. During the next nine weeks, the control, regular diet; HFC, HFC diet, FLU, fluvastatin (3 mg/kg/day); RNL, RN (100 mg/kg/day); RNH, RN (200 mg/kg/day) were orally administered together with the diet, and the experiments were conducted for a total of 13 weeks. The weight of the epididymal adipose tissue, liver, and heart of rats in the HFC diet group significantly increased compared to those in the control group but improved in the RN-treated group. It was also confirmed that vascular function, which is damaged by an HFC diet, was improved after RN treatment. The levels of insulin, glucose, triglycerides, total cholesterol, and low-density lipoprotein increased in the HFC diet group compared to those in the control group, while the administration of RN attenuated these parameters. In addition, the administration of RN significantly reduced the gene expression of both LXR and SREBP-1, which indicated the inhibitory effect of the biosynthesis of triglycerides caused by RN. The results indicated that RN administration resulted in an improvement in the overall lipid metabolism and a decrease in the concentration of triglycerides in the HFC diet-induced rat model of hypertriglyceridemia. Therefore, our findings suggest that the RN can be a candidate material to provide a new direction for treating hypertriglyceridemia.

Мікроскопічне дослідження біосумісності зразків з вмістом мультипотентних мезенхімальних стромальних клітин жирової тканини в умовах експерименту

The purpose of the study was to investigate the biocompatibility of samples containing multipotent mesenchymal stromal cells of adipose tissue to replace bone defects. Material and methods. The study was conducted at Bukovina State Medical University, Chernivtsi, Ukraine. Adipose tissue samples were obtained from the neck of 60 experimental animals (white Wistar rats). We selected 4 samples for the toxicological experiment, which allowed to establish the direct influence of factors in the contact of implantation material at the cellular level. Sample № 1 - Multipotent mesenchymal stromal cells of adipose tissue, which underwent osteogenic differentiation; № 2 - Multipotent mesenchymal stromal cells of adipose tissue with osteogenic differentiation with the addition of platelet-enriched blood plasma; № 3 - “Kolapan” with applied tissue culture of Multipotent mesenchymal stromal cells of adipose tissue cells, which underwent osteogenic differentiation; № 4 - "Kolapan" + Multipotent mesenchymal stromal cells of adipose tissue + platelet-enriched plasma. Multipotent mesenchymal stromal cells of adipose tissue were obtained by grinding adipose tissue of rats in 0.1% collagenase 1A [14]. The study of biocompatibility by cell culture in vitro was performed in accordance with the Working Instruction № 04/2013-VL. The cultures were investigated by the explantation method in a plasma clot in Karelian vials. In order to standardize the nature of growth, their zones were classified into compact, reticular and migrating cells of growing fibroblastic tissues. To assess the probability of the obtained results of the study we used a variation-statistical method of analysis using Microsoft Excel. Statistical calculation of the results of clinical and laboratory studies was carried out according to conventional methods. Results and discussion. Microscopic examination of the surface of samples with culture of fibroblasts showed their satisfactory adhesion on the tooth surface after 5 days of cultivation. In the study of sample № 1 (Multipotent mesenchymal stromal cells of adipose tissue, with osteogenic differentiation), it was noted that the structure of the cells acquired a rounded and oval shape, which indicated their destruction and damage. On the 5th day of observation, cells with numerical intussusception and processes were observed during visual examination of sample № 3 (“Kolapan”, with applied culture of Multipotent mesenchymal stromal cells of adipose tissue). In the study of samples № 4 (Multipotent mesenchymal stromal cells of adipose tissue + platelet-enriched plasma + "Kolapan") on the 5th day of research, signs of growth were manifested by migration of fibroblastic elements that had a spindle-shaped and polygonal shape, with the formation of the primary zone due to strands. On the 7th day of cultivation in experimental samples № 2, № 3, № 4 there was the formation of three growth zones: compact - from cells of polygonal and spindle-shaped form; reticulate - from strands and bundles of cells that were located reticulate and areas of single migrating elements of spindle-shaped. External characteristics and cell growth surface did not differ from control samples. On the 10th day of cultivation in the experimental samples, as well as in the control, the areas of compact and reticular growth zone and the zone of migrating fibroblasts were increased. At the same time, tissue-like growth of cells was observed. Visualization of compact and stack-like zones of the studied experimental samples revealed signs of the beginning of degenerative changes, which was characterized in the form of rounding of the shape and vacuolation of cells. This trend was most pronounced in samples № 2 and № 4. Conclusion. Thus, tissue equivalents of bone tissue based on Multipotent mesenchymal stromal cells of adipose tissue can be candidates for use in regenerative medicine, and studies of their application in experimental animals will provide an opportunity to expand the understanding of the characteristics of Multipotent mesenchymal stromal cells of adipose tissue in order to optimize their further clinical application and implement new approaches in different areas of dentistry