Atorvastatin attenuates the paraquat-induced pulmonary inflammation via PPARγ receptors: a new indication for atorvastatin.

This study was carried out to highlight the role of PPARγ receptors and atorvastatin's protective effect on paraquat (PQ)-induced inflammation in the lungs. Forty-two male Wistar rats were exposed either against saline as control or PQ (3.5 mg/kg, IP) as test groups for 14 days. The test groups were nominated as: PQ, pioglitazone (PGT, 10 mg/kg, orally), atorvastatin (STN, 10 mg/kg, orally), PGT+STN, PGT+GW9662 (1 mg/kg) and STN+GW9662 (1 mg/kg). PGT and STN significantly (P<0.05) reduced the PQ-elevated myeloperoxidase activity, nitric oxide and malondialdehyde contents of the lungs and IL-6 and TNF-α concentrations in serum. Histopathological studies revealed alveolar edema and hemorrhages along with hyaline exudates in alveoli confirming that PGT and STN reduced the damages. Immunohistochemistry studies showed that the PQ-induced inflammation resulted in a severe recruitment of CD68(+) macrophages, which PGT and STN remarkably diminished them. STN regulated the PQ-up-regulated COX-2 expression. The antagonistic effect of GW9662 as an absolute antagonist of PPARγ receptors on anti-inflammatory effect of STN in the regulation of COX-2 expression was observed. These data provide a molecular proof(s) of the STN-produced protective effects on the PQ-induced pulmonary inflammation, which is antagonized by PPARγ antagonist indicating its anti-inflammatory effects via PPARγ receptors. Moreover, a new indication for atorvastatin is suggested.

Vitamin E improved cypermethrin-induced damages in the ovary of rats; evidence for angiogenesis and p53 involvement.

This study aimed to investigate the protective effect of vitamin E (VitE) on cypermethrin (CPM)-induced damages in the ovary. Wistar rats were divided into seven groups (n=6) including; control-sham (c), CPM-received (CPM, 75 mg/kg, i.p.), and CPM and VitE-treated (VitE, 150 mg/kg, orally) for 7, 14 and 24 days. The antioxidant status determination and hormonal assays along with histological and immunofluorescent assessments were performed. The expression of p53 at mRNA level was also examined. The CPM administration affected the ovarian structure and functions as it elevated the follicular atresia and significantly (P<0.05) lowered the estradiol level, time dependently. VitE administration enhanced the CPM-reduced antioxidant capacity, gonadotropins and estradiol levels. Co-administration of VitE and CPM remarkably attenuated the CPM-induced RNA damage in granulosa and theca cells and elevated the deranged angiogenesis. The CPM-reduced micro and macro vessels distribution was significantly (P<0.05) elevated in the VitE-received animals. Expression of p53 at mRNA level was down regulated in the VitE-treated groups completely and relatively following 7 and 14 days, respectively. Our data showed that the CPM-induced biochemical and histological damages could be prevented by VitE. Moreover, protective effects of VitE attribute to its potency in enhancing the antioxidant capacity and promoting the gonadotropins secretion, which resulted in down regulation of p53 overexpression and RNA damage in follicular cells accomplished with improved angiogenesis.

Effects of silymarin on the pharmacokinetics of atorvastatin in diabetic rats.

The effect of silymarin (SMN) on the pharmacokinetics of atorvastatin in diabetic rats was evaluated. Male Wistar rats were assigned into two major groups and then sub-grouped according to the purposes of the study. The first major group was subdivided into three groups (n = 6) including control, non-treated diabetic and SMN-treated diabetic animals. In the first major group, metabolism of testosterone by the hepatic microsomes was studied. The second major group also was divided to three groups including atorvastatin-treated non-diabetic, atorvastatin-treated diabetic and diabetic animals which received both atorvastatin and SMN. To study the pharmacokinetics of atorvastatin, serum samples were collected at 0, 3, 6, 12 and 24 h after the atorvastatin administration. Pharmacokinetic parameters were calculated using non-compartmental model. Streptozotocin-induced diabetes resulted in a remarkable induction of testosterone hydroxylation as the V max for 6ß-hydroxytestosterone production in the diabetic rats (77.3 ± 8.6 pM/min/mg) was significantly higher than that in the control animals (45.9 ± 5.9 pM/min/mg). Moreover, SMN-treated animals showed a significant (P < 0.05) reduction of V max (59.4 ± 6.1 pM/min/mg). Diabetes resulted in a significant reduction of AUC (control 6.98 ± 0.58 vs diabetic rats 4.35 ± 0.24 h mg/ml) and C max values (control 0.52 ± 0.03 vs diabetic group 0.33 ± 0.01 µg/ml), while the SMN-received group showed remarkable recovery of diabetes-reduced values of AUC and C max. These findings indicated that diabetes resulted in a significant up-regulation of microsomal enzyme activities. Moreover, as SMN could significantly regulate the enzyme activities and consequently the atorvastatin pharmacokinetics in diabetic rats, its regulative effect in a combination therapy is concluded.

Evaluation of Effect of Biologically Synthesized Ethanolic Extract of Propolis-Loaded Poly(-Lactic-co-Glycolic Acid) Nanoparticles on Wound Healing in Diabetic Rats.

Wound healing is interaction of a complex cascade of cellular/biochemical actions leading to restoration of structural and functional integrity with regain of injured tissues strength. This study was aimed at evaluation of application of ethanolic extract of propolis-loaded poly(-lactic-co-glycolic acid) nanoparticles (EEP-PLGA NPs) on wound healing in diabetic rats. Sixty rats were randomized into four groups of 15 rats each: In control group (Control) diabetic wound was treated with normal saline. In Carrier 1 group diabetic wound was treated with PLGA nanoparticles based solution. In Carrier 2 group the diabetic wound was treated with EEP. In Treatment group animals received EEP-PLGA NPs on the wound. Wound size was measured on 7, 14 and 21 days after surgery. The expression of p53, bcl-2, Caspase III, were evaluated using reverse-transcription PCR and Immunohistochemical staining. The Treatment group had significantly reduced the wound size compared to other groups (P = 0.001). histological and morphometric studies, and mean rank of the qualitative studies demonstrated that there was significant difference between Treatment group and other groups (P < .05). Observations demonstrated that ethanolic extract of propolis-loaded PLGA nanoparticles significantly shortened the inflammatory phase and accelerated the cellular proliferation. Accordingly, the animals in Treatment group revealed significantly (P < .05) higher fibroblast distribution/one mm2 of wound area and rapid re epithelialization. The mRNA levels of bcl-2, p53 and caspase III were remarkably (P < .05) higher in Treatment group compared to control and animals. The immunohistochemical analyzes confirmed the RT-PCR findings. EEP-PLGA NPs offered potential advantages in wound healing acceleration and improvement through angiogenesis stimulation, fibroblast proliferation and granulation tissue formation in early days of healing phases, acceleration in diabetic wound repair associated with earlier wound contraction and stability of damaged area by rearrangement of granulation tissue and collagen fibers.

Curcumin-Polyethylene Glycol Loaded on Chitosan-Gelatin Nanoparticles Enhances Burn Wound Healing in Rat.

The aim of the present study was to evaluate effects of curcumin-polyethylene glycol loaded on chitosan-gelatin nanoparticles (C-PEG-CGNPs) on burn wound healing in rat as a model study. Sixty healthy male White Wistar rats were randomized into four experimental groups of 15 animals each: Control group (Control) was treated with normal saline. Carrier group was treated with CGNPs-based ointment (0.05 mg/ml). Silver sulfadiazine group was treated with silver sulfadiazine 1% ointment. Treatment group was treated with C-PEG-CGNPs (0.05 mg/ml). Wound size was measured on 7, 14, and 21 days after surgery. The expression of p53, Bcl-2, caspase-3 were evaluated using reverse transcription-polymerase chain reaction and immunohistochemical staining. Reduction in wound area indicated that there was significant difference between Treatment group and other groups (P < .05). Quantitative histological and morphometric studies, and mean rank of the qualitative studies demonstrated that there was a significant difference between Treatment group and other groups (P < .05). Observations demonstrated C-PEG-CGNPs significantly shortened the inflammatory phase and accelerated the cellular proliferation. Accordingly, the animals in Treatment group revealed significantly (P < .05) higher fibroblast distribution/one mm2 of wound area and rapid reepithelialization. The mRNA levels of Bcl-2, p53, and caspase-3 were remarkably (P < .05) higher in Treatment group compared to control animals. The immunohistochemical analyses confirmed the reverse transcription-polymerase chain reaction findings. C-PEG-CGNPs offered potential advantages in burn wound healing acceleration and improvement.

Overexpression of GDNF and FGF-1 in Canine Benign Prostatic Hyperplasia: Evidence for a Pathogenetic Role of Neural Growth Factor.

Benign prostatic hyperplasia (BPH) is common in aged dogs, but the pathogenesis has not been clearly elucidated. A total of 33 male Iranian dogs of mixed breed and in three age groups (under 3 years [n = 10]; 3-6 years [n = 15]; over 6 years [n = 8]), were investigated. BPH was confirmed by ultrasonography and histopathology in 13 cases. The highest prevalence of BPH was in the 3-6 years age group (8/15; 53.3%). Examination of sections of prostate that had been stained with Masson's trichrome revealed that the intensity of stromal smooth muscle cell staining (P <0.05) and the number of fibroblasts (P = 0.002) were significantly increased in BPH compared with normal prostate glands. Prostate cells from dogs with BPH (n = 13) had a significantly higher intensity of cytoplasmic immunolabelling with antibodies against glial cell line-derived neurotrophic factor (GDNF), cytokeratin (CK) AE1/AE3, vimentin, fibroblast growth factor-1 (FGF-1) and prostate-specific antigen (PSA), compared with normal prostate glands (n = 20) (P = 0.001), except for PSA, which was negative in both normal and BPH affected prostates. The overexpression of GDNF and FGF-1 in stromal and epithelial cells of prostate glands of dogs with BPH suggests that GDNF has a paracrine or autocrine role in stimulating cellular proliferation. GDNF overexpression may also play a pathogenetic role in promoting chronic prostatitis and increasing fibrosis and the smooth muscle component of the prostate gland in BPH.

Evaluation of antibiotic activity of methicillin in healing of full-thickness infected wounds with sensitized methicillin resistant Staphylococcus aureus in the presence of HAMLET.

OBJECTIVES: The novel healing choices for handling of infections due to multidrug resistant Staphylococcus aureus are reguired. HAMLET has been reported to be able to sensitize bacterial pathogens to traditional antimicrobial agents. The aim was to assess wound healing activity of methicillin in presence of HAMLET in methicillin resistant S. aureus (MRSA) infected wounds. MATERIALS AND METHODS: Fifty male rats were randomized into five groups of ten animals each. In CONTROL group, 0.1 ml sterile saline 0.9% solution was added to the wounds with no infection. In MRSA group, the wounds were infected with MRSA and only treated with 0.1 ml the sterile saline (0.9%) solution. In MRSA/HAMLET group, infected wounds were cured with HAMLET (100 µg). In group MRSA/ Met, animals with infected wounds were cured with 0.1 ml local use of 1 mg/ml methicillin. In MRSA/Met/HAMLET group, animals with infected wounds were cured with local use of 0.1 ml solution of methicillin (1 mg/ml) and HAMLET (100 µg). All test formulations were used for ten consecutive days, twice a day, beginning from first treatment. RESULTS: Microbiological examination, planimetric, histological and quantitative morphometric studies, immunohistochemical staining for angiogenesis, determination of hydroxyproline levels and RT-PCR for Caspase 3, Bcl-2 and p53 showed that there was significant difference between animals in MRSA/Met/ HAMLET group compared to other groups (P<0.05). CONCLUSION: HAMLET could make methicillin beneficial for handling of MRSA infected wounds and had the prospective effect to consider this harmless agent for local application.

Functional, Histopathological and Immunohistichemical Assessments of Cyclosporine A on Sciatic Nerve Regeneration Using Allografts: A Rat Sciatic Nerve Model.

OBJECTIVES: To study the functional, histopathological and immunohistochemical effect of cyclosporine A on sciatic nerve regeneration using allografts in a rat sciatic nerve model. METHODS: Thirty male white Wistar rats were divided into three experimental groups (n = 10), randomly: Normal control group (NC), allograft group (ALLO), CsA treated group (ALLO/ CsA). In NC group left sciatic nerve was exposed through a gluteal muscle incision and after homeostasis muscle was sutured. In the ALLO group the left sciatic nerve was exposed through a gluteal muscle incision and transected proximal to the tibio-peroneal bifurcation where a 10 mm segment was excised. The same procedure was performed in the ALLO/ CsA group and the animals were treated with interaperitoneal administration of cyclosporine A. The harvested nerves of the rats of ALLO group were served as allograft for ALLO/ CsA group and vice versa. The NC and ALLO groups received 300 µL sterile olive oil interaperitoneally once a day for one week and the ALLO/ CsA group received 300 µL CsA (1mg/kg/day) interaperitoneally once a day for one week. RESULTS: Behavioral, functional, biomechanical and gastrocnemius muscle mass showed earlier regeneration of axons in ALLO/ CsA than in ALLO group (p=0.001). Histomorphometic and immunohistochemical studies also showed earlier regeneration of axons in ALLO/ CsA than in ALLO group (p=0.034). CONCLUSION: Administration of CsA could accelerate functional recovery after nerve allografting in sciatic nerve. It may have clinical implications for the surgical management of patients after nerve transection in emergency conditions.

Co-Administration of Vitamin E and Testosterone Attenuates The Atrazine-Induced Toxic Effects on Sperm Quality and Testes in Rats.

OBJECTIVE: Atrazine (ATZ) as a widely used herbicide is considered as a potent endocrine disrupter which adversely affects reproductive systems in both genders. This study aimed to assess the effects of testosterone (T)- and vitamin E (VitE)- alone and their coadministration on testicular function and sperm parameters after exposure to ATZ in rats. MATERIALS AND METHODS: In this experimental study, the rats (n=30) are assigned into the following 5 groups: control-sham group (n=6) receiving corn oil, ATZ group (n=6) receiving 200 mg/kg ATZ alone, ATZ+VitE group (n=6) receiving 150 mg/kg ATZ+VitE, ATZ+T group (n=6) receiving 400 µg/kg ATZ+T, and ATZ+VitE+T group (n=6) receiving ATZ+VitE+T for 48 consecutive days. Total antioxidant capacity (TAC), total thiol molecules (TTM), and malondialdehyde (MDA) were analyzed. Serum levels of T, luteinizing hormone (LH), and inhibin-B (IN-B) were also determined. Histological examination and sperm analysis were performed. The data were analyzed using Graph-Pad Prism software version 2.01. RESULTS: Co-administration of VitE and T significantly (P<0.05) increased ATZ-decreased TAC and TTM levels and reduced ATZ-increased MDA content. T and VitE significantly (P<0.05) increased serum levels of ATZ-reduced T (1.94 ± 0.96), IN-B (122.10 ± 24.33) and LH (0.40 ± 0.10). The T+VitE animals showed a reduction in apoptotic cells and an increase in Leydig cells steroidogenesis. Co-administration of T and VitE significantly (P<0.05) reduced the ATZ-induced DNA disintegrity and chromatin de-condensation. VitE and T protected germinal cells RNA and protein contents against ATZ-induced damages. CONCLUSION: T and VitE in simultaneous form of administration were able to normalize the ATZ-induced derangements through promoting antioxidant capacity and endocrine function.

Silymarin: A Novel Natural Agent to Restore Defective Pancreatic ß Cells in Streptozotocin (STZ)-induced Diabetic Rats.

This study aimed to investigate the potency of silymarin (SMN) and melatonin (MEL) on restoring the pancreatic   cells in streptozotocin (STZ)-induced diabetic rats. Male Wistar rats were divided into five groups, including: control (C), untreated diabetic (D), SMN-treated diabetic (50 mg/Kg, orally), MEL-treated diabetic (10 mg/Kg, i.p.), and SMN plus MEL-treated diabetic rats. Diabetes was induced by injection of STZ (50 mg/Kg, i.p.). The blood glucose and insulin levels were measured. After the 28 days treatment period, antioxidant status was analyzed by determination of total antioxidant capacity (TAC) in the liver and serum. The histopathological changes in the pancreatic islets were examined by histochemical staining and enumeration of   cells. Although none of the test compounds reduced the blood glucose level to normal concentration, however SMN alone and in combination with MEL was able to decline it significantly (P<0.05) after 28 days administration. Both SMN and MEL could recover the diabetes-reduced TAC values. Moreover, the diabetes-induced cellular vacuolation and   cells depletion were improved by the SMN treatment. Our data suggest that the SMN and MEL treatment was able to normalize the antioxidant status, while only SMN administration could restore the  cells of Langerhans islets in diabetic rats.

Effect of phosalone on testicular tissue and in vitro fertilizing potential.

BACKGROUND: The current study aimed to evaluate the effects of phosalone (PLN) as an organophosphate (OP) compound on testicular tissue, hormonal alterations and embryo development in rats. MATERIALS AND METHODS: In this experimental study, we divided 18 mature Wistar rats into three groups-control, control-sham and test (n=6 per group). Animals in the test group received one-fourth the lethal dose (LD50) of PLN (150 mg/kg), orally, once per day for 45 days. DNA laddering and epi-fluorescent analyses were performed to evaluate testicular DNA fragmentation and RNA damage, respectively. Serum levels of testosterone and inhibin-B (IN-B) were evaluated. Testicular levels of total antioxidant capacity (TAC), total thiol molecules (TTM) and glutathione peroxidase (GSH-px) were analyzed. Finally, we estimated sperm parameters and effect of PLN on embryo development. Two-way ANOVA was used for statistical analyses. RESULTS: There was severe DNA fragmentation and RNA damage in testicular tissue of animals that received PLN. PLN remarkably (p<0.05) decreased testicular TAC, TTM and GSH-px levels. Animals that received PLN exhibited significantly (p<0.05) decreased serum levels of testosterone and IN-B. Reduced sperm count, viability, motility, chromatin condensation and elevated sperm DNA damage were observed in the test group rats. PLN resulted in significant (p<0.05) reduction of in vitro fertilizing (IVF) potential and elevated embryonic degeneration. CONCLUSION: PLN reduced fertilization potential and embryo development were attributed to a cascade of impacts on the testicles and sperm. PLN promoted its impact by elevating DNA and RNA damages via down-regulation of testicular endocrine activity and antioxidant status.

Allelic frequency and genotypes of prion protein at codon 136 and 171 in Iranian Ghezel sheep breeds.

PrP genotypes at codons 136 and 171 in one hundred twenty Iranian Ghezel sheep breeds were studied using allele-specific PCR amplification and compared with the well-known sheep breeds in North America, the United States, and Europe. The frequency of V allele and VV genotype at codon 136 of Ghezel sheep breed was significantly lower than AA and AV. At codon 171, the frequency of allele H was significantly lower than Q and R. Despite the similarities of PrP genotypes at codons 136 and 171 between Iranian Ghezel sheep breeds and some of the studied breeds, significant differences were found with others. Planning of effective breeding control and successful eradication of susceptible genotypes in Iranian Ghezel sheep breeds will not be possible unless the susceptibility of various genotypes in Ghezel sheep breeds to natural or experimental scrapie has been elucidated.