Protective effect of crocin against reperfusion-induced cardiac arrhythmias in anaesthetized rats

The aim of the present study was to investigate the effects of crocin - a natural antioxidant derived from saffron - on cardiac reperfusion-induced arrhythmia and antioxidant systems such as catalase and superoxide dismutase (SOD) enzyme activities, glutathione (GSH) and malondialdehyde (MDA, as a marker of lipid peroxidation) levels. Rats in 4 experimental groups were administered crocin (20 mg/kg/day) or vehicle (i.p.) for 21 days with or without cardiac ischemia-reperfusion (IR). At the end of this period, hearts of anaesthetized animals in IR and “Cr + IR” groups were subjected to 10 min occlusion of the left anterior descending coronary artery and thereafter reperfused for 30 min. The results suggest that crocin is partially capable of suppressing reperfusion-induced arrhythmias. Compared to control group, ischemic-reperfusion injury significantly decreased SOD activity and GSH level and increased MDA level of heart muscle. “Cr + IR” group showed remarkably increased catalase activity in heart tissue (28.7 ± 6.6 vs. 23.6 ± 4.1 U/mg protein, P < 0.05) compared to the IR group. The level of cardiac tissue SOD activity in the “Cr + IR” group animals did not decline significantly compared to rats that were administered crocin alone with no ischemia. The results suggest a protective role of crocin on cardiac reperfusion arrhythmias which may at least partially be related to stability or even amplification of antioxidant systems. Crocin may potentially be useful for treatment or prevention of arrhythmias in patients with ischemic heart disease and this issue remains to be investigated in future clinical studies

The effect of high intensity interval training on cardioprotection against ischemia-reperfusion injury in Wistar rats

The aims of the present study were to determine whether short term high intensity interval training (HIIT) could protect the heart against ischemia reperfusion (IR) injury; and if so, to evaluate how long the exercise-associated protection can be lasted. Sixty-three rats were randomly assigned into sedentary (n = 15), sham (n = 7), and exercise groups (n = 41). Rats in the exercise groups performed 5 consecutive days of HIIT on treadmill: 5 min warm up with 50 % VO2max, 6×2min with 95-105 % VO2max (about 40 to 45 m/min), 5×2 min recovery with 65-75 % VO2max (about 28 to 32 m/min), and 3 min cool down with 50 % VO2max, all at 0 % grade. Animals exposed to an in vivo cardiac IR surgery, performed at days 1, 7, and 14 following the final exercise session. Ischemia-induced arrhythmias, myocardial infarct size (IS), plasma lactate dehydrogenase (LDH) and creatine kinase (CK) activities were measured in all animals. Compared to sedentary rats, exercised animals sustained less IR injury as evidenced by a lower size of infarction and lower levels of LDH and CK at day one and day 7 post exercise. In comparison of sedentary group, IS significantly decreased in EX-IR1 and EX-IR7 groups (50 and 35 %, respectively), but not in EX-IR14 group (19 %). The exercise-induced cardioprotection disappeared 14 days following exercise cessation. There were no significant changes in ischemia-induced arrhythmia between exercised and sedentary rats. The results clearly demonstrate that HIIT protects the heart against myocardial IR injury. This protective effect can be sustained for at least one week following the cessation of the training

Cloning and expression of human vasohibin1 gene in E. coli

Background: Angiogenesis is an important process in various physiologic and pathologic states. The most significant stimulator of angiogenesis is vascular endothelial growth factor (VEGF). In contrast, vasohibin1 acts as an angiogenesis inhibitor which specifically inhibits new vessels formation. The aim of the present study was cloning and expression of vasohibin1 gene in E. coli as well as purification of recombinant vasohibin1 protein. Methods: Total RNA was extracted from human umbilical vein endothelial cells and cDNA was synthesized by RT-PCR. cDNA was amplified using a specific designed primer set. The PCR product was evaluated by electrophoresis and then cloned in pET28a expression vector which transformed into E.coli BL21 (DE3) as a host. IPTG is used as an expression inducer in media. Alternatively, PCR products were analyzed by sequencing and double digestion with EcoRI and HindIII restriction endonuclease. The expressed protein was purified by Ni-NTA column and confirmed by SDS Page and western blotting. Evaluation of gene inhibition was carried out through western blottting and RT-PCR. Results: No mutation or sequence variants were found in PCR products as a result of sequencing analysis. Moreover, the quantity and quality of expressed recombinant protein in the presence of IPTG with selected vector in E. coli was approximately high. VASH1 significantly prevented the receptor expression. The quality and level of expressed protein in pET28 expression vector indicated the efficacy of the applied system in vasohibin1 production. Conclusions: The produced vasohibin1 protein probably can be used as an angiogenesis inhibitor in further studies on retinopathies

Cloning and expression of human vasohibin1 gene in E. coli

Background: Angiogenesis is an important process in various physiologic and pathologic states. The most significant stimulator of angiogenesis is vascular endothelial growth factor (VEGF). In contrast, vasohibin1 acts as an angiogenesis inhibitor which specifically inhibits new vessels formation. The aim of the present study was cloning and expression of vasohibin1 gene in E. coli as well as purification of recombinant vasohibin1 protein. Methods: Total RNA was extracted from human umbilical vein endothelial cells and cDNA was synthesized by RT-PCR. cDNA was amplified using a specific designed primer set. The PCR product was evaluated by electrophoresis and then cloned in pET28a expression vector which transformed into E.coli BL21 (DE3) as a host. IPTG is used as an expression inducer in media. Alternatively, PCR products were analyzed by sequencing and double digestion with EcoRI and HindIII restriction endonuclease. The expressed protein was purified by Ni-NTA column and confirmed by SDS Page and western blotting. Evaluation of gene inhibition was carried out through western blottting and RT-PCR. Results: No mutation or sequence variants were found in PCR products as a result of sequencing analysis. Moreover, the quantity and quality of expressed recombinant protein in the presence of IPTG with selected vector in E. coli was approximately high. VASH1 significantly prevented the receptor expression. The quality and level of expressed protein in pET28 expression vector indicated the efficacy of the applied system in vasohibin1 production. Conclusions: The produced vasohibin1 protein probably can be used as an angiogenesis inhibitor in further studies on retinopathies

Sodium metabisulfite-induced changes on testes, spermatogenesis and epididymal morphometric values in adult rats

Background: Sulphites are widely used as a preservative and antioxidant additives in the food and pharmaceutical industries. Many types of biological and toxicological effects of sulphites in multiple organs of mammals have been shown in previous studies. Objective: The aim of this study was to investigate the effects of sodium metabisulfite (SMB) on testicular function and morphometric values of epididymis in adult male Wistar rats. Materials and Methods: A total of 32 rats were randomly divided into four groups. The experimental groups received SMB at doses of 10 mg/kg (S10), 100mg/kg (S100), and 260 mg/kg (S260) while an equal volume of normal saline was administered to the control group via gavage. The rats were anaesthetized after 28 days and the left testis with the head of epididimis was excised following abdominal incision for histological observation using hematoxylin and eosin staining. Serum samples were collected for assay of testosterone level. The initial epididymis was analyzed for motility, morphology, and the number of sperms. Result: The results of this study showed that normal morphology, count, and motility of sperms and testosterone level were decreased in the SMB treated groups. In comparison with the control group, SMB resulted in a lower total number of spermatogonia, primary spermatocyte, spermatids, and Leydig cells. Conclusion: It is suggested that SMB decreases the sperm production and has the potential to affect the fertility adversely in male rats

The effects of Artemisia aucheri extract on hepatotoxicity induced by thioacetamide in male rats

Objective: Liver is an important organ that is exposed to many oxidant and carcinogenic agents, thus antioxidant compounds are beneficial for liver health. Artemisia contains flavonoid compounds and anti-diabetic, antioxidant, and anti-inflammatory properties. Due to possessing terpene and sesquiterpene compounds, this plant has antioxidant properties. This study was done to investigate the effects of Artemisia plant extract on thioacetamide-induced hepatotoxicity in Wistar rats. Materials and Methods: For induction of hepatotoxicity, 50 mg/kg thioacetamide was injected intraperitoneally (i.p).After extraction and purification, the hydroalcoholicextract was injected i.p. at 100, 200, and 300 mg/kg doses for 21 days together with thioacetamide at 50 mg/kg dose in the last 3 days. After blood sampling and separation of serum, alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), albumin, and total protein concentrations were measured. Results: Significant decreases in aminotransferase and alkaline phosphatase activities and significant increases in the concentration of albumin and total protein in groups treated with the extract compared with thioacetamide-treated group were observed (

The effects of co-administration of opium and morphine with nicotine during pregnancy on spatial learning and memory of adult male offspring rats

Objective(s): Smoking opium/cigarette is a global health concern. The aim of this study was to examine learning and memory of rat male offsprings whose mothers had been exposed to either opium or morphine with nicotine during pregnancy. Materials and Methods: Wistar rats were used for the experiments. In the female rats, opium, morphine and nicotine dependencies were induced by daily injections of drug solution for 10 days before mating. Spatial memory was tested by Morris water maze test in male pups at the postnatal day 60. The duration that took until the rats found the platform in the maze and also their swimming speed were recorded. Results: An increase in the platform finding duration was observed for the pups of dependent mothers in comparison with the control in the training trial (