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Recently, substantial attention has been paid toward adipose-derived mesenchymal stem cells (AdMSCs) as a potential therapy in tissue engineering and regenerative medicine applications. Rat AdMSCs (r-AdMSCs) are frequently utilized. However, the influence of the adipose depot site on the multilineage differentiation potential of the r-AdMSCs is still ambiguous. Hence, the main objective of this study was to explore the influence of the adipose tissue harvesting location on the ability of r-AdMSCs to express the stem-cell-related markers and pluripotency genes, as well as their differentiation capacity, for the first time. Herein, we have isolated r-AdMSCs from the inguinal, epididymal, peri-renal, and back subcutaneous fats. Cells were compared in terms of their phenotype, immunophenotype, and expression of pluripotency genes using RT-PCR. Additionally, we investigated their potential for multilineage (adipogenic, osteogenic, and chondrogenic) induction using special stains confirmed by the expression of the related genes using RT-qPCR. All cells could positively express stem cell marker CD 90 and CD 105 with no significant in-between differences. However, they did not express the hematopoietic markers as CD 34 and CD 45. All cells could be induced successfully. However, epididymal and inguinal cells presented the highest capacity for adipogenic and osteogenic differentiation (21.36-fold and 11.63-fold for OPN, 29.69-fold and 26.68-fold for BMP2, and 37.67-fold and 22.35-fold for BSP, respectively, in epididymal and inguinal cells (p < 0.0001)). On the contrary, the subcutaneous cells exhibited a superior potential for chondrogenesis over the other sites (8.9-fold for CHM1 and 5.93-fold for ACAN, (p < 0.0001)). In conclusion, the adipose tissue harvesting site could influence the differentiation capacity of the isolated AdMSCs. To enhance the results of their employment in various regenerative cell-based therapies, it is thus vital to take the collection site selection into consideration.

This study assessed the effects of a simulated high-altitude environment on the reproductive system of prepubertal male rats and the reversibility of these effects upon return to a normal environment. Three-week-old male Wistar rats were randomly allocated to 4 groups that were exposed to different conditions: a normal environment for 6 weeks and 12 weeks, respectively, hypobaric hypoxia for 6 weeks, and hypobaric hypoxia for 6 weeks followed by a normal environment for 6 weeks. Multiple pathophysiological parameters were evaluated at the histological, endocrine, and molecular levels. Hypobaric hypoxia exposure for 6 weeks during the prepubertal phase significantly altered physiological parameters, body functions, blood indices, and reproductive potential. Six weeks after returning to a normal environment, the damaged reproductive functions partially recovered due to compensatory mechanisms. However, several changes were not reversed after returning to a normal environment for 6 weeks, including disorders of body development and metabolism, increased red blood cells, increased fasting blood glucose, abnormal blood lipid metabolism, decreased testicular and epididymis weights, abnormal reproductive hormone levels, excessive apoptosis of reproductive cells, and decreased sperm concentration. In summary, a hypobaric hypoxic environment significantly impaired the reproductive function of prepubertal male rats, and a return to normal conditions during the postpubertal phase did not fully recover these impairments.

Diabetes can affect male fertility via oxidative stress and endocrine system disruption. Nanomedicine based on natural products is employed to address diabetes complications. The current study aims to investigate the potential beneficial effect of propolis extract nanoparticles against diabetes-induced testicular damage in male rats. Sixty male rats were randomly allocated to six groups ( n  = 10). The first group served as a control group. The second and third received propolis extract (Pr) and propolis extract nanoparticles (PrNPs). The fourth group is the diabetic group that received streptozotocin (STZ) (55 mg kg/bwt) single-dose i/p. The fifth and sixth groups are diabetic rats treated with Pr and PrNPs. Both Pr and PrNPs were received at a dose (100 mg/kg bwt) orally. After 60 days, animals were euthanized, then pancreatic and testicular tissues were collected for redox status evaluation, gene expression analysis, and histopathological examination. Also, hormonal analysis (Insulin, total testosterone, and luteinizing hormone (LH) ) along with semen quality evaluation were done. Results showed that the induction of diabetes led to testicular and pancreatic redox status deterioration showing a reduction in reduced glutathione (GSH) as well as elevation of malondialdehyde (MDA), and nitric oxide (NO) levels. Also, relative transcript levels of testicular cytochrome P450 family 11 subfamily A member 1 (CYP11A1) , 3β-Hydroxysteroid dehydrogenase (HSD-3β) , and nuclear factor (erythroid-derived 2)-like 2 (NFE2L2) were significantly down-regulated, While the advanced glycation end-product receptor (AGER) relative gene expression was significantly upregulated. Furthermore, hormonal and semen analysis disturbances were observed. Upon treatment with Pr and PrNPs,  a marked upregulation of testicular gene expression of CYP11A1 , HSD-3β , and NFE2L2 as well as a downregulation of AGER, was observed. Hormones and semen analysis were improved. In addition, the testicular and pancreatic redox status was enhanced. Results were confirmed via histopathological investigations. PrNPs outperformed Pr in terms of steroidogenesis pathway improvement, testicular antioxidant defense mechanism augmentation, and prospective antidiabetic activity.

Summary Osteocyte is the orchestrator of bone remolding and decline in osteocyte autophagy is involved in senile osteoporosis. Our results suggested that rapamycin, at least in part by activating osteocyte autophagy, reduced the severity of age-related bone changes in trabecular bone of old male rats. Introduction Previous literatures have showed that osteocyte is the orchestrator of bone remolding and age-related decline in osteocyte number is associated with senile osteoporosis. Autophagy is an important cellular protective mechanism which can preserve osteocyte viability and failure of autophagy in osteocyte with age has been linked to senile osteoporosis. The purpose of this study was to explore whether rapamycin, one activator of autophagy, has protective effects on senile osteoporosis through inducing osteocyte autophagy. Methods Fifty-two 24-month-old male Sprague-Dawley (SD) rats were randomly divided into two groups. Rapamycin (1 mg/kg weight/day) or DMSO vehicle control was administered intraperitoneally for 12 weeks. BMD and bone microstructure were determined by Micro-CT. Fluorochrome labeling of the bones was performed to measure the mineral apposition rate (MAR). TRAP staining was performed to evaluate osteoclast number. The plasma levels of bone turnover markers were also analyzed. The effects of rapamycin on osteocyte autophagy were determined by immunohistochemistry, Western blot, and q-PCR. TUNEL was used to determine the prevalence of osteocyte apoptosis. Results Micro-CT evaluation demonstrated that rapamycin had a protective effect on age-related bone loss in trabecular bone. Besides, rapamycin resulted in an obvious increase of MAR and a decrease of osteoclast number in contrast to the control group. Furthermore, rapamycin also induced autophagy in osteocyte demonstrated by increased LC3-positive osteocyte and increased LC3 turnover. In addition, rats treated with rapamycin exhibited decreased apoptosis of osteocyte determined by TUNEL. Conclusions These results suggested that rapamycin, at least in part by activating osteocyte autophagy, reduced the severity of age-related bone changes in trabecular bone of old male rats. Therefore, rapamycin might be a feasible therapeutic approach for senile osteoporosis.

Adolescence is a key period of development when major cognitive and neurobiological changes occur. Results from our lab showed that 5 days of social isolation in adolescent rats led to molecular changes in the Wnt/β-catenin pathway and to higher cocaine responses during adulthood. We assessed whether 5 days of social isolation (SI) during adolescence would impact on β-catenin levels in the prefrontal cortex (PFC) and nucleus accumbens (NAcc) as well as on anxiety-like behaviors in a sex- and time-dependent manner. We also investigated the role of dopaminergic neurotransmission on that impact, by using repeated administration of a D2 antagonist. Male and female Wistar rats were socially isolated between postnatal day (PND)30 to 35 or kept in their home cages (non-isolated), while they were treated with sulpiride (100 mg/kg, ip ) or vehicle. Anxiety-like behaviors and exploratory activity were estimated by the open field test at 24 h (PND36) or 9 days (PND44) after isolation. Then, they were euthanized at PND36 or PND45, and β-catenin levels were analyzed by Western blot in PFC and NAcc. Our findings show that a brief SI during adolescence leads to a long-term impact on both β-catenin levels (10 days, PND45) and anxiety-like behaviors (9 days, PND44) with a significant increase and decrease, respectively, in female rats. In contrast, male rats show a rapid decrease in β-catenin levels in the PFC with no changes in anxiety-like behaviors (24 h, PND36). These suggest that adolescent SI induces mostly long-term changes in female while short term changes in male rats. Moreover, these changes seem to be modulated by dopaminergic neurotransmission since a sulpiride treatment during isolation prevented them.

Swimming is a popular sport with several health benefits, but its effects on bone quality are controversial possibly due to distinct effects on different anatomical regions. Our aim was to investigate the effect of 8-month swimming on bone growth, mass, geometry, trabecular microarchitecture and osteocyte density of the lumbar vertebrae, femur and tibia of male rats. Wistar rat models were assigned to either a swimming ( n  = 10; 2h/d, 5 d/week) or a physically active control group ( n  = 10) for 8 months, after which they were sacrificed and their lumbar vertebrae, femur and tibia assessed for bone mass, cortical geometry, trabecular microarchitecture and osteocyte density through µ -CT and histology. Variables were compared between groups through independent samples t tests. Swimming animals displayed higher vertebral trabecular connectivity and lower trabecular separation compared to controls. However, femur length, trabecular and cortical bone mass and cortical thickness were lower compared to controls. At the tibia, animals from the swimming group also presented lower trabecular number and connectivity and higher trabecular separation. Osteocyte density at the femur and vertebra was similar between groups. Eight months of swimming negatively affected bone mass, cortical geometry and trabecular microarchitecture at the femur and tibia whilst having a favourable effect on vertebral trabecular microarchitecture. These results suggest that swimming has divergent effects on different anatomical regions.

BackgroundIn a previous study in female rats, voluntary wheel running attenuated incubation of cocaine craving after 30 but not 3 days (Zlebnik and Carroll Zlebnik and Carroll, Psychopharmacology 232:3507–3413, 2015). The present study in male rats, using the same procedure, showed that wheel running reduced incubated craving after both 30 and 3 days of abstinence.MethodsMale rats self-administered i.v. cocaine (0.4 mg/kg) during 6-h sessions for 10 days. They were then moved from the operant chamber to a home cage with an attached running wheel or stationary wheel, for 6 h daily for a 3- or 30-day period when cocaine craving was hypothesized to incubate. Rats were then returned to the operant chamber for a 30-min test of cocaine seeking, or “craving,” indicated by responses on the former “drug” lever was formerly associated with drug stimulus lights and responses (vs. no drug stimuli), and lever responding was compared to responses on the “inactive” that was illuminated and counted lever pressing.ResultsMean wheel revolutions were similar across the 3- and 30-day incubation groups, when both groups of rats were given access to wheel running vs. access to a stationary wheel in controls. Subsequently, when rats were tested in the operant chamber for “relapse” responding (drug-lever responding) on the lever formerly associated with drug access, cocaine craving was reduced by recent running wheel access (vs. stationary wheel access) in both the 3- and 30-day wheel exposure groups.ConclusionVoluntary, self-initiated, and self-sustained physical exercise reduced cocaine craving after short- (3 days) and long-term (30 days) abstinence periods in male rats that previously self-administered cocaine. This was contrasted with reduction of cocaine seeking in females after 30-day, but not 3-day, incubation periods under the wheel running vs. stationary wheel conditions in a previous study (Zlebnik and Carroll Zlebnik and Carroll, Psychopharmacology 232:3507–3413, 2015). These initial findings suggest males may be more sensitive to incubated craving for cocaine than females.

Recently, the world has been dealing with diseases that spread easily and weaken the body’s natural defenses. Boosting natural immunity can help prevent these diseases. This study sought to assess the immunomodulatory effects of extracts from Illicium verum , Saussurea costus , and Glycyrrhiza glabra , both individually and in conjunction with chitosan and carboxy chitosan nanoparticles (CS NPs and C.CS NPs), in immunosuppressed male rats. The primary objective was to assess whether the combined plant extracts exhibit superior immunomodulatory effects compared to single treatments. The herbal extracts were tested chemically. The nanoparticles and loaded herbal extracts were studied for size, charge, and chemical structure, and their safety was tested on two cell types. The study used 104 adult male rats, divided into 13 groups: a control group, an immunosuppressed group, three groups treated with herbal extracts, two nano groups, and six groups treated with loaded herbal extracts. Immunosuppression was caused by injecting 20 mg of dexamethasone weekly. Herbal extracts, CS NPs, C.CS NPs, and the loaded herbal extracts were given orally every day after day for 28 days. The study checked immune health, tissue structure, and cell growth marker Ki-67. All treatments improved immune health by increasing white blood cells, complement 3 and 4, interferon-gamma, and tumor necrosis factor levels, and raised Ki-67 expression. Incorporation of chitosan and methyl carboxy chitosan (C.Cs) nanoparticles amplified the immunostimulatory effect at a reduced dosage of 100 mg/kg, with the C.Cs-based formulation exhibiting enhanced therapeutic performance. The results highlight the promise of nanoparticle-formulated herbal extracts, especially C.CS-loaded variants, as potent immunomodulatory agents with enhanced therapeutic efficacy.

The present study aimed to reveal the effects of immunocastration on the development of the immune system in rats. Seventy rats were randomly assigned into two groups: Control (n = 35) and immunized (n = 35). Twenty-day-old rats were immunized with gonadotropin-releasing hormone (GnRH) and booster immunization was administered every two weeks (three immunizations in total). From 20-day-old rats, we collected samples every two weeks, including five immunized rats and five control rats (seven collections in total). We collected blood samples, testicles, thymuses, and spleens. The results showed that GnRH immunization increased the GnRH antibody titers and reduced the testosterone concentration (both P < 0.05 ). Compared with the control group, the number of CD4+CD8− cells, CD4−CD8+ cells, and CD4+CD8+ cells increased ( P < 0.05 ) whereas the number of CD4-CD8- cells and CD4+CD25+ cells reduced in the immunized group ( P < 0.05 ) over time. GnRH immunization also increased the relative weights of thymus and spleen ( P < 0.05 ), serum concentrations of interleukin (IL)-2, IL-4, IL-6, IL-10, IL-17 and Interferon-γ (IFN-γ) over time ( P < 0.05 ), and changed the mRNA levels of IL-2, IL-4, IL-6. IL-10, IL-17, IFN-γ, CD4, D8, CD19 GnRH, and GnRH receptor ( GnRH-R ) in thymus and spleen. Thus, GnRH immunization enhanced the immune markers in thymus, spleen, and blood immune cytokines in rats.

In this study, molecular interactions of the ligands, quercetin, gallic acid, and metformin with various diabetes mellitus-related protein targets, such as glycogen phosphorylase and peroxisome proliferator-activated receptor gamma, were assessed. It was revealed that quercetin possesses good binding affinity to both targets. Quercetin is a major constituent of methanolic extracts of Phyllanthus emblica fruit. The antihyperglycemic effect of quercetin in streptozotocin (STZ)-induced diabetic rats was examined. The isolated quercetin administered at a dose of 75mg/kg body weight produced a maximum decrease of 14.78% in blood glucose levels in the diabetic rats after 7 days of treatment. Furthermore, quercetin doses of 50 and 75mg/kg were shown to significantly improve the profiles of triglycerides, high-density lipoprotein, very-low-density lipoprotein, low-density lipoprotein, and total cholesterol at the end of the study in STZ-induced diabetic rats. The administration of quercetin (25, 50, and 75mg/kg body weight) daily for 28 days in STZ-induced diabetic rats resulted in a significant decrease in blood glucose and urine sugar levels, with a considerable rise in plasma insulin and hemoglobin levels. Therefore, quercetin is a potential drug with antidiabetic and antihyperglycemic action mediated by changes in the levels of glucose, cholesterol, and triglycerides as indicated by in silico and in vivo studies.