Comparative effects of thymol and vitamin E on nonalcoholic fatty liver disease in male Wistar rats.

Following the obesity epidemics, nonalcoholic fatty liver disease (NAFLD) has grown in prevalence and become a main cause of morbidity and death, intimately linked to cardiovascular disease, cancer, and cirrhosis. The key factor in the evolution of NAFLD is thought to be oxidative stress. Because most patients cannot change their lifestyle or dietary habits, a pharmaceutical strategy is now required to treat NAFLD. Nonalcoholic fatty liver disease (NAFLD), including nonalcoholic steatohepatitis, is treated with vitamin E. (NASH). Vitamin E is also a powerful antioxidant that has been demonstrated to lower oxidative stress in people with NAFLD. Thymol is a monoterpene phenol with a variety of pharmacological effects, however its anti-fatty liver properties have yet to be investigated. Despite the fact that oxidative stress is thought to have a role in the etiology of nonalcoholic steatohepatitis, antioxidant therapies have not been well studied in the treatment of nonalcoholic steatohepatitis. The goal was to learn more about vitamin E and thymol's biological activities, with a particular emphasis on their therapeutic effectiveness in NAFLD. Four groups of thirty-two adult male rats were formed (healthy control, thymol, Vit E, and fatty liver). For 28 days, rats were given either oral vitamin E (200 mg/kg) or thymol (50 mg/kg) randomly. The levels of ALT, AST, TNF- α, Ferritin, CK-MB enzymes, and MAPK gene expression were then determined in the serum. Based on a random effect model analysis, at the end of 28 days of therapy, ALT (41.43 U/L), AST (47.91 U/L), Ferritin (1.13 pg/dl), CK-MB (251.22 IU/L), TNF-α (95.39 pg/mL) (p≤0.001), and MAPK gene expression levels (p≤0.05) significantly reduced in both experimental groups compared with the fatty liver group. Vitamin E and thymol therapy is a safe, affordable, and effective therapeutic option in the fatty liver group. Patients with fatty liver disease should be encouraged to take vitamin E and Thymol supplements, which are both safe and affordable, because more effective new therapeutic options are lacking.

Artemisia absinthium improves spatial performance and neuronal injury induced by amyloid- beta in the CA1 hippocampal area of male Wistar rats.

Alzheimer's disease is a neurodegenerative disorder. It is characterized by the presence of two aberrant structures in the brain, those are, amyloid plaques and neurofibrillary tangles, along with neuronal death. Amyloid-beta further exacerbates the metabolic decline and results in cognitive impairments. Because of the favorable antioxidant and anti-inflammatory activities of Artemisia absinthium (wormwood), this study aimed to evaluate the effects of hydroalcoholic extract of this plant on spatial memory performance, neuronal injury, and apoptosis induced by amyloid-beta. Forty-eight male Wistar rats (220-250 g) were divided into the following groups: 1) control; 2) sham (solvent; ICV); 3) amyloid-beta 1-40 (ICV); and 4) amyloid-beta plus A. absinthium (10, 50, and 100 mg/kg/day; gavage). Congo red and TUNEL staining were performed to investigate the neuronal injury. Also, the Morris water maze (MWM) test was used to evaluate the spatial memory of the experimental groups. The results showed that spatial memory for finding the hidden platform in the MWM task decreased significantly in the amyloid-beta group, compared to the control and sham groups. In contrast, treatment with A. absinthium improved spatial memory dose-dependently and reduced tissue degeneration, amyloid plaques, and apoptosis. It seems that the hydroalcoholic extract of A. absinthium, due to its antioxidant and anti-inflammatory activities, can effectively reverse spatial memory deficits and reduce amyloid-beta plaques.

Neuroprotective Effects of Normobaric Hyperoxia and Transplantation of Encapsulated Choroid Plexus Epithelial Cells on The Focal Brain Ischemia.

OBJECTIVE: Choroid plexus epithelial cells (CPECs) have the epithelial characteristic, produce cerebrospinal fluid, contribute to the detoxification process in the central nervous system (CNS), and are responsible for the synthesis and release of many nerve growth factors. On the other hand, studies suggest that normobaric hyperoxia (HO) by induction of ischemic tolerance (IT) can protect against brain damage and neurological diseases. We examined the effect of combination therapy of encapsulated CPECs and HO to protect against ischemic brain injury. MATERIALS AND METHODS: In this experimental study, six groups of adult male Wistar rats were randomly organized: sham, room air (RA)+middle cerebral artery occlusion (MCAO), HO+MCAO, RA+MCAO+encapsulated CPECs, HO+MCAO+encapsulated CPECs, RA+MCAO+empty capsules. RA/HO were pretreatment. The CPECs were isolated from the brain of neonatal Wistar rats, cultured, and encapsulated. Then microencapsulated CPECs were transplanted in the neck of the animal immediately after the onset of reperfusion in adult rats that had been exposed to 60 minutes MCAO. After 23 hours of reperfusion, the neurologic deficit score (NDS) was assessed. Next, rats were killed, and brains were isolated for measuring brain infarction volume, blood-brain barrier (BBB) permeability, edema, the activity of superoxide dismutase (SOD), and catalase (CAT) and also, the level of malondialdehyde (MDA). RESULTS: Our results showed that NDS decreased equally in HO+MCAO, RA+MCAO+encapsulated CPECs, and HO+MCAO+encapsulated CPECs groups. Brain infarction volume decreased up 79%, BBB stability increased, edema decreased, SOD and CAT activities increased, and MDA decreased in the combination group of HO and transplantation of encapsulated CPECs in the ischemic brain as compared with when HO or transplantation of encapsulated CPECs was applied alone. CONCLUSION: The combination of HO and transplantation of encapsulated CPECs for stroke in rats was more effective than the other treatments, and it can be taken into account as a promising treatment for ischemic stroke.

Preparation and Evaluation of A Novel Liposomal Nano-Formulation in Metastatic Cancer Treatment Studies.

OBJECTIVE: Today, in clinical trials, we suffer from the lack of effective methods with minimal side effects to deliver medication. Thus, efforts to identify better conditions for delivery of biomedical drugs seem necessary. The purpose of this study was to design a new liposomal formula for transportation of microRNA in osteosarcoma. MATERIALS AND METHODS: In this experimental study, several liposomal formulations were synthesized. Physical and chemical parameters, including size, zeta potential, polydispersity index, long-term stability of the liposomal-microRNA complex and the amount of miR-143 loading in liposome based nano-vesicles were optimized using different techniques. Similarly, the effect of free and encapsulated microRNA toxicity were investigated and compared in a human bone osteosarcoma cell line, named SaOs-2. RESULTS: In this study, we could produce a novel and optimized formulation of cationic PEGylated liposomal microRNA for gene delivery. The present synthesized microRNA lipoplex system was non-agglomerated. The system remained stable after four months and miR-143 leakage was not observed by performing gel electrophoresis. The microRNA lipoplex could enhance conduction of the loaded miR-143, and it also showed good biocompatibility to the healthy cells. CONCLUSION: The PEGylated microRNA lipoplex system had a high potential for the systematic migration of miR-143 and it could improve intracellular stability of the released microRNA.

Neuroprotective Effects of Combined Treatment with Minocycline and Olfactory Ensheathing Cells Transplantation against Inflammation and Oxidative Stress after Spinal Cord Injury.

OBJECTIVE: Traumatic spinal cord injury (SCI) is considered one of the most devastating injuries leading to neuronal disruption. Olfactory ensheathing cells (OECs) and minocycline have been shown to promote locomotor function after spinal cord injury. In this study, we have tested the efficacy of combined treatment with minocycline and OECs after contusive spinal cord injury. MATERIALS AND METHODS: In this experimental study, adult female Wistar rats were randomly divided into five groups. Rats received an intraperitoneal injection of minocycline immediately after SCI, and then 24 hours after the injury. Transplantations were performed 7 days after the injury. Functional recovery was evaluated using the Basso, Beattie and Bresnahan scale (BBB). After that, the animals were sacrificed, and T11 segment of the spinal cord was removed after 5 weeks, and then used for histopathological, immunohistochemical, and biochemical assessments. Western blot analysis was applied to determine the protein expression of tumor necrosis factor alpha (TNF-α), interleukin 1 beta (IL1ß) and caspase3. RESULTS: The results of this study showed that the combination of OECs graft and minocycline reduced the functional deficits and diminished cavitation and astrogliosis in spinal tissue. The analysis of protein expression by western blotting revealed that minocycline treatment along with OECs transplantation further decreased the level of IL-1ß, TNF-α, caspase-3, and the oxidative stress as compared with when minocycline or OECs transplantation was used alone. CONCLUSION: The combinatory treatment with OECs graft and minocycline induced a more effective response to the repair of spinal cord injury, and it is considered a therapeutic potential for the treatment of SCI.

Tissue Engineered Human Amniotic Membrane Application in Mouse Ovarian Follicular Culture.

Since folliculogenesis requires a powerful cell-matrix interaction, natural scaffolds seem to be needed for follicular culture. Human amniotic membrane (HAM) offers promise as a support of in vitro ovarian follicular culture. HAM was decellularized with trypsin and EDTA. DNA and histology assays were performed to determine the elimination rate of genomic components. Cyto-biocompatibility of decellular AM (DAM) was verified by the cell viability (MTT) test. The small parts of intact amniotic membrane (IAM) and DAM were coated on the bottom of 96-well and each well was filled with 150 µL of base medium. Mouse primary-secondary (PS) follicles were separated to three groups: 1-culture in base medium (Control), 2-culture on IAM and 3-culture on DAM. Follicular size, morphology, viability, estradiol production and genes expression were evaluated and IAM group showed better growth and development in follicle culture. The viability rate and estradiol production in both experimental groups were statistically higher than the Control. Gdf9, Bmp15 and Cx37 were found to have higher expression levels in IAM group. Also, maximum apoptotic and survival indexes were determined in Control and IAM groups, respectively. Finally, IAM provides a better protective environment for mouse PS follicular culture that can reduce apoptosis level.

ASSOCIATION OF TESTOSTERONE WITH COLORECTAL CANCER (HT29), HUMAN GLIOBLASTOMA (A172) AND HUMAN EMBRYONIC KIDNEY (HEK293) CELLS PROLIFERATION.

Testosterone influences cancer development. This in vitro experiment was exerted to determine the association of testosterone with human colorectal cancer(HT29), glioblastoma (A172) and human embryonic kidney(HEK293) cells proliferation. HT-29, A172 and HEK293 cell lines were cultured in standard growth medium, then randomly divided into control group (not exposed to testosterone) and groups exposed to 1, 10, 100 and 1000 µg/mL of testosterone. Cell viability was quantified by MTT assay. Statistical analysis was performed using ANOVA. Viability of HEK293 cells significantly increased in groups exposed to 1 µg/mL and decreased in groups exposed to 100 and 1000 µg/mL of testosterone compared to control group (P<0.05, P<0.05 and P<0.001, respectively). Viability of HT29 cells significantly increased in groups exposed to 10 and 100 µg/mL of testosterone and significantly decreased when exposed to 1000 µg/mL of testosterone compared to control group (P<0.05, P<0.001 and P<0.001, respectively). Viability of A172 cells significantly decreased in groups exposed to 100 and 1000 µg/mL of testosterone compared to control group (P<0.001). In conclusion, different doses of testosterone have enhancing or suppressive effects on HEK293, HT29 and A172 cells proliferation; according to which, considering clinical use of testosterone therapy for cancer treatment is a highly controversial issue.

Role of Mas receptor antagonist (A779) in renal hemodynamics in condition of blocked angiotensin II receptors in rats.

The vasodilatory effect of angiotensin 1-7 (Ang 1-7) is exerted in the vascular bed via Mas receptor (MasR) gender dependently. However, the crosstalk between MasR and angiotensin II (Ang II) types 1 and 2 receptors (AT1R and AT2R) may change some actions of Ang 1-7 in renal circulation. In this study by blocking AT1R and AT2R, the role of MasR in kidney hemodynamics was described. In anaesthetized male and female Wistar rats, the effects of saline as vehicle and MasR blockade (A779) were tested on mean arterial pressure (MAP), renal perfusion pressure (RPP), renal blood flow (RBF), and renal vascular resistance (RVR) when both AT1R and AT2R were blocked by losartan and PD123319, respectively. In male rats, when AT1R and AT2R were blocked, there was a tendency for the increase in RBF/wet kidney tissue weight (RBF/KW) to be elevated by A779 as compared with the vehicle (P=0.08), and this was not the case in female rats. The impact of MasR on renal hemodynamics appears not to be sexual dimorphism either when Ang II receptors were blocked. It seems that co-blockade of all AT1R, AT2R, and MasR may alter RBF/ KW in male more than in female rats. These findings support a crosstalk between MasR and Ang II receptors in renal circulation.

Role of Mas receptor antagonist (A779) on pressure diuresis and natriuresis and renal blood flow in the absence of angiotensin II receptors type 1 and 2 in female and male rats.

Sexual differences in blood pressure are associated with angiotensin 1-7 (Ang1-7) and its receptor and enzyme function targeting. Blockade of angiotensin II (AngII) receptors type 1 and 2 (AT1R and AT2R) inhibits some actions of Ang1-7. We described the role of Ang1-7 receptor (MasR) antagonist (A779) on kidney hemodynamics when AT1R and AT2R are blocked with losartan and PD123319. In anaesthetized male and female rats after blockade of both AT1R and AT2R, the renal perfusion pressure (RPP) was controlled in two levels of 80 and 100 mmHg via an adjustable clamp placed around the aorta above the level of the renal arteries. Then, the effects of saline vehicle and MasR blocker (A779) were tested on pressure natriuresis and diuresis, renal blood flow (RBF), and renal vascular resistance (RVR). In the absence of AT1R and AT2R; RVR, RBF/wet kidney tissue weight, and serum level of renin did not alter in both genders either MasR was blocked or not. However, urine flow rate (UF) and sodium excretion (UNaV) increased significantly at the pressure level of 100 mmHg in the presence of MasR in male (P<0.05) but not in female rats. When AT1R and AT2R were blocked, the impact of MasR is gender-related in pressure natriuresis and diuresis, and pressure natriuresis and diuresis in male rats (not female) increases in the presence of MasR.

Effect of caloric restriction on nitric oxide production, ACE activity, and blood pressure regulation in rats.

High calorie intake and high weight gain is one of the worldwide health problems particularly in industrial and developed countries. The subjected individuals are at high risk for developing various disorders such as diabetes and particularly cardiovascular problems. It has been well established that life style modification plays an important role in reducing these problems, particularly weight reduction and caloric restriction (CR) as a non-pharmacological approach. This study sought to examine the possible effect of caloric restriction on nitric oxide production, ACE activity and blood pressure regulation in rat. Two groups of rats were selected as the control (C) and the CR group and a with standard and an every other day diet, respectively, for 4 weeks. At the end of study in the CR group systolic blood pressure was significantly decreased compared to controls. The serum NOx was significantly increased compared to the C group. The serum ACE activity was lower in the CR group. Therefore, it may be concluded that CR could reduce blood pressure by elevating NO production and lowering ACE activity.

Effect of a new phenanthroline derivative on testicular tissue and serum testosterone concentration and its contraceptive affects on adult Balb/C mouse strain.

In this research, the effect of 2, 6-diaminopyridinum as a new phenanthroline derivative was studied on the hypophysis-gonad axis, testicular tissue and sperm production in male Balb/C mice. Fifty adult male Balb/C mice were divided in five groups. First group was considered as untreated control. Saline was injected to second group and the remaining three groups received intraperitoneal injection of 15, 20 and 25 mg kg(-1) of 2, 6-diaminopyridinum every other day for 20 days. The LD50 was determined to be 35 mg kg(-1) body weight. The testicular tissues were studied morphologically and the serum concentration of FSH, LH and testosterone were measured. The results showed that 25 mg kg(-1) diaminopyridinum decreased the number of germ cells significantly and serum testosterone level with no change on FSH and LH levels. This study indicates that 25 mg kg(-1) of phenanthroline may directly affect testicular tissue causing a lower testosterone level and spermatogenesis in mice.

Baclofen-impairment of memory retention in rats: possible interaction with adrenoceptor mechanism(s).

This study concerned the influence of adrenoceptor agonists and antagonists on baclofen-induced impairment of memory retention. Intracerebroventricular injection of baclofen (0.25--2 microg/rat) reduced memory retention in rats. The combination of different doses of baclofen with a low dose of clonidine (0.5 microg/rat) elicited a greater decrease in memory retention. Yohimbine (1 microg/rat) potentiated the response to a low dose, but decreased the response to higher doses of baclofen. Single administration of clonidine (0.5--2 microg/rat) but not yohimbine (1--4 microg/rat) itself decreased memory retention. The combination of clonidine with yohimbine did not show any interaction. The low dose of phenylephrine (0.5 microg/rat) or prazosin (0.5 microg/rat) also potentiated the inhibition of memory retention by baclofen. Phenylephrine (0.5--3 microg/rat) increased, while prazosin (0.5--2 microg/rat) decreased memory retention. The combination of the two drugs showed an interaction. It may be concluded that an adrenoceptor mechanism may interact with the memory retention impairment induced by baclofen.