The effects of G protein-coupled receptor 30 (GPR30) on cardiac glucose metabolism in diabetic ovariectomized female rats.

BACKGROUND: Diabetic cardiometabolic disorders are characterized by significant changes in cardiac metabolism and are increased in postmenopausal women, which emphasize the role of 17ß-estradiol (E2). Despite this, there are few safe and effective pharmacological treatments for these disorders. The role of G protein-coupled estrogen receptor (GPR30), which mediates the non-genomic effects of E2, is mostly unexplored. METHODS: In this study, we used ovariectomy (menopausal model) and type 2 diabetic (T2D) rats' models to evaluate the preclinical action of G-1 (GPR30 agonist) against cardiometabolic disorders. T2D was induced by a high-fat diet and a low dose of streptozotocin. G-1 was administrated for six weeks after the establishment of T2D. RESULTS: We found that G-1 counteracts the effects of T2D and ovariectomy by increasing the body weight, reducing fasting blood sugar, heart weight, and heart weight to body weight ratio. Also, both ovariectomy and T2D led to decreases in the cardiac protein levels of hexokinase 2 (HK2) and GLUT4, while G-1-treated female rats reversed these changes and only increased HK2 protein level. In addition, T2D and ovariectomy increased glucose and glycogen content in the heart, but G-1 treatment significantly reduced them. CONCLUSIONS: In conclusion, our work demonstrates that G-1 as a selective GPR30 agonist is a viable therapeutic approach against T2D and cardiometabolic diseases in multiple preclinical female models.

Ferulic Acid Ameliorates Cell Injuries, Cognitive and Motor Impairments in Cuprizone-Induced Demyelination Model of Multiple Sclerosis.

OBJECTIVE: Ferulic acid (FA) is a phenolic compound that exhibits neuroprotective effects in the central nervous system (CNS). This study was conducted to evaluate the potential effects of FA on the cognitive and motor impairments in the cuprizone-induced demyelination model of multiple sclerosis (MS). MATERIALS AND METHODS: In this experimental study, demyelination was induced in mice by feeding them with chow containing cuprizone (CPZ) 0.2% for 6 weeks. Mice in the control group received normal chow. Mice in the CPZ+Veh, CPZ+FA10, and CPZ+FA100 groups received saline, and FA at a dose of 0, 10, or 100 mg/kg (intraperitoneal, I.P., daily) respectively. After cognitive and motor assessments, under anaesthesia, animal brains were removed for evaluating the histological, apoptosis, and molecular changes. RESULTS: The results showed that FA increased freezing behaviour in contextual (P<0.05) and cued freezing tests (P<0.05). FA also reduced the random arm entrance (P<0.01) and increased spontaneous alternations into the arms of Y-maze compared to the CPZ+Veh group (P<0.05). Time on the rotarod was improved in rats that received both doses of FA (P<0.01). Demyelination, apoptosis, and relative mRNA expression of p53 were lower in the FA-treated groups relative to the CPZ+Veh group (P<0.01). In addition, FA increased mRNA expression of brain-derived neurotrophic factor (Bdnf), Olig2, and Mbp (P<0.05) but decreased GFAP mRNA expression compared to the CPZ+Veh group (P<0.01). CONCLUSION: The results of this study showed that FA plays a significant neuroprotective role in CPZ models of demyelination by reducing neuronal apoptosis and improving oligodendrocytes (OLs) growth and differentiation.

Progesterone eliminates 17ß-estradiol-Mediated cardioprotection against diabetic cardiovascular dysfunction in ovariectomized rats.

BACKGROUND: Type2 Diabetes (T2D) remains one of the most important causes of cardiovascular diseases (CVD). Menopause leads to an increase in CVD and metabolic syndrome, which indicates the role of sex steroids as a protective factor. In the present study, we surveyed the effects of 17ß-estradiol (E2) alone and in combination with progesterone (P4) on cardiovascular dysfunction in T2D. METHODS: Female ovariectomized (OVX) diabetic rats were divided into eight groups: Sham-Control, Diabetes (Dia), OVX + Dia, OVX + Dia + Vehicle, OVX + Dia + E2, OVX + Dia + P4, OVX + Dia + E2+P4, and OVX + Dia + E2+Vehicle. T2D was induced by a high-fat diet and streptozotocin. E2 and P4 were administrated every four days for four weeks. The heart cytokines and angiotensin II, lipid profile, insulin, water, and food intake and cardiovascular indices were measured. RESULTS: Results showed that single treatment with E2 decreased fasting blood glucose, water, and food intake, atherogenic and cardiac risk indices, and blood pressure. Also, P4 led to a decrease in atherogenic and cardiac risk indices. TNFα and IL-6 levels were increased and IL-10 was decreased in the Dia group, while E2 alone was able to inhibit these changes. The combined use of E2 and P4 eliminated the beneficial effects of E2 on these indices. Although diabetes results in an increment of cholesterol, LDL and triglyceride, hormone therapy with E2 was associated with improved dyslipidemia. CONCLUSION: The use of E2 alone, and not the individual use of P4, and its combination with E2 improved cardiovascular function in OVX diabetic animals, possibly by reducing the amount of inflammatory cytokines and improving metabolic parameters.

Role of the potassium channels in vasorelaxant effect of asafoetida essential oil.

OBJECTIVE: In a previous work, we showed that asafoetida essential oil (AEO), from oleo-gum resin of Ferula asafoetida L. from the Apiaceae family, has a vasodilatory effect. This effect was both endothelium-dependent and endothelium-independent. The present study was designed to determine whether potassium channels and intracellular calcium release contribute to AEO-induced vasodilation. MATERIALS AND METHODS: Rats' thoracic aorta were isolated and denuded. Following induction of contraction by potassium chloride (60 mM), concentration-response curve was plotted by the cumulative addition of AEO (0.625-80 µl/l to the medium of rings. The vasodilatory effect of AEO was assessed before and after addition of phenylephrine and potassium channel blockers (including barium chloride (BC), 4-aminopyridine (4A) and glibenclamide (Gl)). RESULTS: AEO relaxed the precontracted rings in a concentration-dependent manner (IC50=23 µl/l). All potassium channel blockers significantly attenuated the vasodilatory activity of AEO when they were added to rings medium before addition of KCl (p<0.01, 4A and Gl groups and p< 0.001, BC group vs. control group) but not after that. In contrast to K channel blockers, adding AEO before or after phenylephrine, the tension was reduced significantly (p<0.05 vs. the control group). CONCLUSION: The findings of this study indicated that the vasodilatory effect of AEO on denuded-endothelium aortic ring was mediated through activation of potassium channels and reduced intracellular calcium release.

17ß-Estradiol improves insulin signalling and insulin resistance in the aged female hearts: Role of inflammatory and anti-inflammatory cytokines.

Aging effects in energy balance in all tissues and organs, including the cardiovascular. The risk of cardiovascular disease is drastically higher in postmenopausal women than in premenopausal women. Estrogen plays an important role in the cardiac function and body's metabolism. The aim of this study was to determine whether 17ß-estradiol (E2) has beneficial effects on insulin resistance and some key stages of the insulin signalling pathway in the aged hearts. Young and aged female Wistar rats were ovariectomized and were randomly divided into three groups: young (YS) and aged (AS) sham, young (YV) and aged (AV) vehicle, and young (YE2) and aged (AE2) E2 treatment groups. E2 (1 mg/kg) was administrated every four days for four weeks. Results showed that ovariectomy increased fasting blood glucose, insulin, and HOMAIR in young, while none of these parameters was affected in aged animals. On the other hand, aging itself increased these variables. Furthermore, E2 therapy alleviated these changes in both young and aged animals. Moreover, aging also decreased the p-IRS1, p-Akt level, and translocation of GLUT4 to the plasma membrane. E2 reduced the negative impact of menopause and aging on insulin sensitivity by favoring increase in the level of IL-10 and decrease in the levels of TNF-α and IL-1ß. Our results indicated that the heart response to E2 depended on age, and E2 increased insulin sensitivity in the heart of both young and aged animals by altering inflammatory conditions. Determining the exact mechanism of this action is suggested in future studies.

Ischemic Preconditioning Efficacy Following Anabolic Steroid Usage: A Clear Difference Between Sedentary and Exercise-Trained Rat Hearts.

Previous studies show that anabolic steroids impair innate cardioprotective mechanisms. Here, we investigated the effect of supraphysiological doses of nandrolone on ischemic preconditioning (IPC) as a potent cardioprotective tool against ischemia reperfusion (IR) injury in rat hearts. Male Wistar rats in two experimental settings of sedentary and exercise-trained (60 min/day swimming, 5 days/week, for 8 weeks) were either pretreated with intramuscular injections of arachis oil (Arach, n = 16) as vehicle or nandrolone decanoate (ND, n = 8), 10 mg/kg/week, for 8 weeks. At the end, the hearts were excised and perfused in a Langendorff system. Then, the vehicle-treated hearts subdivided into the IR (30 min of LAD coronary artery occlusion and 120 min reperfusion, n = 8) and IPC (three cycles of 3-min ischemia and 3-min reperfusion before test ischemia, n = 8) groups and nandrolone-treated hearts served as ND + IPC (nandrolone pretreatment before IR and IPC protocols, n = 8) group. Post-ischemic cardiac function and infarct size were assessed. Reperfusion arrhythmias were analyzed using a standard scoring system. In sedentary hearts, ND slightly increased heart-to-body weight ratio and increased baseline cardiac contractile function. In trained hearts, ND markedly increased heart-to-body weight ratio which was also associated with enhanced baseline cardiac function. ND pretreatment enhanced protective effects of IPC in sedentary group; however, abolished these effects in exercise-trained group. The arrhythmia score was not significantly different between nandrolone-treated groups vs. respective preconditioned groups. Our findings show that ND impairs IPC-induced cardioprotection in exercise-trained rat hearts. Cardiac hypertrophy seems to play a crucial role in this response.

The effect of asafoetida essential oil on myocardial ischemic-reperfusion injury in isolated rat hearts.

OBJECTIVE: Previous studies reported that asafetida from Ferula assa-foetida Linn. species and its essential oil (AEO) have antioxidant effects. In the present study, the effect of AEO was evaluated on ischemic-reperfusion injury in isolated rat hearts. MATERIALS AND METHODS: Forty-eight male Wistar rats were divided into 6 groups: 1) control group, 2) vehicle group, 3-5) AEO groups and, 6) carvedilol group. In the control group, hearts were only subjected to 30-min global ischemia followed by 120-min reperfusion. Hearts in other groups were perfused with vehicle (Tween 0.1%), AEO (0.125, 0.25 or 0.50 µL/g heart) or carvedilol (10 µM) for 5 min immediately before the induction of ischemia. RESULTS: Compared to the control group, myocardial dysfunction was significantly more severe only in group 5 in which a significant increase in left ventricular end diastolic pressure and a significant decrease in left ventricular developed pressure and ± dp/dt. Also, the activities of lactate dehydrogenase and creatine kinase as the markers of myocardial injury were significantly higher only in group 5 compared to control group. The size of infarct and the incidence of irreversible fibrillation did not show any significant differences between the control group and groups 3-5. CONCLUSION: These results showed that perfusion of isolated rat hearts with AEO 0.5 µL/g heart, but not at lower concentrations, might worsen myocardial ischemic-reperfusion injury.

Cardioprotective and anti-inflammatory effects of G-protein coupled receptor 30 (GPR30) on postmenopausal type 2 diabetic rats.

Diabetic cardiomyopathy is the most common chronic disease in postmenopausal women, but the mechanism(s) is unclear. G-protein coupled receptor 30 (GPR30) is one of the receptors that binds to 17-ß Estradiol (E2). To date, there is little information on GPR30 and its expression in postmenopausal type 2 diabetes (T2D) in the heart. The current study hypothesized that GPR30 mediated cardioprotective effects of E2 in ovariectomized diabetic rats. Female ovariectomized diabetic rats were divided in nine groups: Control, Vehicle, Diabetes, Proestrous, Non-proestrous, E2, E2+Vehicle, E2+G15, and G1. G15 is a GPR30 antagonist, while G1 is an agonist of GPR30. T2D was induced by high fat diet and streptozotocin. E2, G1 and G15 were administrated for four weeks after establishment of T2D. Results showed that mean arterial pressure, fasting blood glucose and HOMA-IR in diabetic and vehicle groups were alleviated by E2 and G1, while salutary effects of E2 were inhibited by G15. Furthermore, E2 and G1 improved cardiac weight, atherogenic and cardiovascular risk indices; meanwhile G15 exacerbated cardiac weight and atherogenic indices. Also, diabetes increased cardiac levels of tumor necrosis factor-alpha and interleukin 6 and E2 only decreased interleukin 6. Significant decrement in the level of interleukin 10, and GPR30 protein were observed in diabetic group, whereas E2 and G1 increased the cardiac levels of interleukin 10, and GPR30 protein. Our study suggested that beneficial and anti-inflammatory effects of E2 on diabetic cardiomyopathy are probably mediated via non-genomic E2 pathways.

Vasodilatory effect of asafoetida essential oil on rat aorta rings: The role of nitric oxide, prostacyclin, and calcium channels.

BACKGROUND: Asafoetida is an oleo-gum resin mainly obtained from Ferula assa-foetida L. species in the apiaceae family. Previous studies have shown that it has antispasmodic effects on rat's and pig's ileums. PURPOSE: The main goals of this study were to assess the vasodilatory effect of asafoetida essential oil (AEO) on the contractile response of rat's aorta rings and to find the role of nitric oxide, cyclooxygenase, and calcium channels. Thoracic aorta rings were stretched under a steady-state tension of 1 g in an organ bath apparatus for 1 h and then precontracted by KCl (80 mM) in the presence and absence of AEO. L-NAME (blocker of nitric oxide synthase) and indomethacin (blocker of cyclooxygenase) were used to assess the role of nitric oxide (NO) and prostacyclin in the vasodilatory effect of AEO. Also, the effect of AEO on the influx of calcium through the cell membrane calcium channels was determined. RESULTS: Data showed that AEO had vasodilatory effects on aorta rings with both intact (IC50 = 1.6 µl/l) or denuded endothelium (IC50 = 19.2 µl/l) with a significantly higher potency in intact endothelium rings. The vasodilatory effects of AEO were reduced, but not completely inhibited, in the presence of L-NAME or indomethacin. Adding AEO to the free-calcium medium also significantly reduced the CaCl2-induced contractions. CONCLUSION: The results indicated that AEO has a potent vasodilatory effect that is endothelium-dependent and endothelium-independent. Also, it reduced the influx of calcium into the cell through plasma membrane calcium channels.

Effect of oleuropein on myocardial dysfunction and oxidative stress induced by ischemic-reperfusion injury in isolated rat heart.

BACKGROUND: Studies have reported antioxidant effect of oleuropein in isolated rat heart. OBJECTIVE: This study was conducted to investigate whether perfusion of isolated rat heart with oleuropein, before induction of ischemia or at the onset of reperfusion, had any effect on the hemodynamic parameters, infarct size and biochemical factors following ischemic - reperfusion injury. MATERIALS AND METHODS: Forty-eight male Wistar rats were divided into 6 groups: the control groups (Con-P and Con-T groups), O10-P and O50-P groups perfused with 10 and 50 µg/g heart oleuropein 5 min before the induction of ischemia and O10-T and O50-T groups perfused with 10 and 50 µg/g heart oleuropein at the beginning of the reperfusion, respectively. All hearts were subjected to 30 min global ischemia and 90 min reperfusion. Hemodynamic parameters were monitored throughout the experiment. The creatine kinase (CK) and malondialdehyde (MDA) level of coronary outflow were assayed and the infarct size measured at the end of reperfusion. RESULTS: We found hemodynamic parameters namely heart rate, left ventricular end diastolic pressure (LVEDP), left ventricular developed pressure (LVDP), ±dp/dt and coronary outflow significantly improved in all groups that received oleuropein compared to the control groups. Also, the infarct size was smaller and the coronary outflow levels of CK and MDA were lower in the oleuropein groups compared to the control groups. CONCLUSIONS: The findings suggest that perfusion of isolated rat heart with oleuropein would lead to improved myocardial dysfunction following ischemic-reperfusion injury. Our findings confirm the antioxidant potential of oleuropein.

Prophylactic and Therapeutic Effects of Oleuropein on Reperfusion-Induced Arrhythmia in Anesthetized Rat.

BACKGROUND: This study was conducted to reveal that whether i.v. injection of oleuropein, the most potent polyphenolic antioxidant in olive leaf, has any effect on the magnitude of reperfusion arrhythmia in anesthetized rats or not. METHODS: Eighty male Wistar rats were divided into 8 groups of 10 each: groups 1 and 5 were assigned as the prophylactic and treatment control groups, groups 2 and 6 as the prophylactic and treatment groups with lidocaine (10 mg/kg), groups 3 and 4 as the prophylactic groups with 10 and 50 mg/kg oleuropein (i.v.), and groups 7 and 8 as the treatment groups with 10 and 50 mg/kg oleuropein (i.v.), respectively. Reperfusion injury was induced by 5-min regional ischemia and 15-min reperfusion of left anterior descending coronary artery. Heart rate, blood pressure, and electrocardiogram were monitored throughout the procedure. RESULTS: blood pressure was significantly decreased by infusion of 50 mg/kg oleuropein in groups 4 and 8, but unlike the lidocaine as a standard anti-arrhythmic drug in groups 2 and 5 had not significant effect on heart rate. The onset of arrhythmia in groups received oleuropein (groups 3, 4, 7, and 8) was significantly delayed. The mortality rate due to irreversible ventricular fibrillation was also significantly reduced in groups 3, 4, 7, and 8. The effect of lidocaine in groups 2 and 5 was more potent than that in oleuropein group. CONCLUSION: These findings indicate that i.v. injection of oleuropein possibly through its antioxidant activity reduces the magnitude of reperfusion-induced arrhythmia.

Hyperoxia-induced preconditioning against renal ischemic injury is mediated by reactive oxygen species but not related to heat shock proteins 70 and 32.

OBJECTIVE: Pre-exposure of rats to normobaric hyperoxia (O2 ≥ 95%) may induce late preconditioning against renal ischemia-reperfusion (IR) injury. In this study we investigated probable mechanisms of IR injury such as the role of reactive oxygen species (ROS), renal antioxidant agents, and heat shock proteins (HSP) 32 and 70 during delayed hyperoxia-preconditioning (HO). METHODS: Fifty-two rats were divided into 7 groups: (A) IR, (B) HO + IR, (C) mercaptopropionyl glycine (MPG) + HO + IR, (D) MPG + IR, (E) HO + sham, (F) MPG + sham, and (G) sham. Rats in the following study groups (group B, C and E) were kept in a normobaric hyperoxic environment for 4 h/day for 6 consecutive days, after which they were subjected to 40 minutes of ischemia; animals in the control group (group A, D, F, and G) were kept in a normoxic cage. At the end of the preconditioning period, 24 hours of reperfusion was performed. Renal function was assessed by measuring serum creatinine (Cr), blood urea nitrogen (BUN), and creatinine clearance (CLCr). Induction of the antioxidant system was evaluated by measuring renal catalase (CAT) and superoxide dismutase (SOD) activities and glutathione (GSH) and malondialdehyde (MDA) content. The role of ROS was investigated by use of MPG (a ROS scavenger). HSP32 & 70 mRNA and protein also were determined. RESULTS: The hyperoxia-preconditioned IR group (B) had a lower plasma Cr and BUN and greater CLCr compared with the IR group (A) (P ≤ .016). Administration of MPG led to an increase in plasma Cr and BUN and a decrease in CLCr in group C compared with the hyperoxia-preconditioned group B (P ≤ .004). The hyperoxia-preconditioned IR group had a greater CAT activity and GSH level compared with the IR group A (P ≤ .007), whereas the administration of MPG did not change the GSH level but led to a decrease in CAT activity in group D compared with group B (P < .001). SOD activity did not change in hyperoxia-preconditioned ischemic rats compared with ischemic rats. Hyperoxia preconditioning and MPG administration in ischemic animals did not result in any considerable change in MDA level compared with the IR group A. Also, there were no clinically relevant differences in HSP32 & 70 mRNA and protein between all groups. CONCLUSION: The present study demonstrates that repeated pre-exposure to hyperoxia can decrease subsequent renal IR damage in this rat model of renal ischemia. Free radical production after hyperoxia appears to play a pivotal role in the hyperoxia-induced renal protection independent of HSP level. Antioxidant enzyme activities and especially catalase seem to be implicated in this renal protective mechanism.

Effect of Ascorbic Acid on Noise Induced Hearing Loss in Rats.

INTRODUCTION: After presbycusis, noise-induced hearing loss is the second most common cause of acquired hearing loss. Numerous studies have shown that high-intensity noise exposure increases free radical species; therefore, use of antioxidants to detoxify the free radicals can prevent cellular damage in the cochlea. We studied the potential hearing protective effect of different doses of ascorbic acid administered prior to noise exposure in rats. MATERIALS AND METHODS: Twenty-four male albino Wistar rats were randomly allocated into four groups: groups A, B, and C received 1250, 250, and 50 mg/kg/day of ascorbic acid, respectively, and group D acted as the control group. After 14 days of ascorbic acid administration, the rats were exposed to noise (105 dB sound pressure level for 2 h). Distortion product otoacoustic emissions (DPOAE) were recorded prior to starting the ascorbic acid as baseline and 1 h after the noise exposure. RESULTS: The amplitude decrease was 14.99 dB for group A, 16.11 dB for group B, 28.82 dB for group C, and 29.91 dB for the control group. Moderate and high doses of ascorbic acid significantly reduced the transient threshold shift in the rats. CONCLUSION: The results of present study support the concept of cochlea protection by antioxidant agents. This dose-dependent protective effect was shown through the use of ascorbic acid treatment prior to noise exposure.

Pretreatment with Assafoetida exerts dose-dependent dual effects on rat hearts.

CONTEXT: Although many studies displayed the favorable effects of Assafoetida, some of them reported that high doses of Assafoetida could lead to harmful effects. AIMS: In this study, the effect of pretreatment with Assafoetida investigated on ischemic-reperfusion injury in isolated rat heart model. MATERIALS AND METHODS: Thirty two male Wistar rats were divided into 4 groups of eight. Group 1 as the control (Con) group and three other groups as the treatment groups that given Assafoetida by gavage at levels of 25, 50 and 100 mg/kg, once a day for four weeks (T25, T50 and T100 groups). Then their hearts were subjected to 30 min global ischemia and 90 min reperfusion under langendorff apparatus. RESULTS: The data shown that hemodynamic parameters including left ventricular developed pressure (LVDP) and maximum and minimum of pressure changes (±dp/dt) were increased in T25 and decreased in T50 and T100 groups during reperfusion in comparison with Con group. There was not any significant difference in the incidence of irreversible ventricular fibrillation between T25 and Con group, while it was increased in T50 and T100 groups significantly. There was not any significant difference in infarct size between all groups. CONCLUSION: These data indicate that pretreatment of rats with Assafoetida have cardioprotective effects in low doses and cardiotoxic effects in higher doses. Therefore, it needs more investigation in the future.

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 anti-infarct, antistunning and antiarrhythmic effects of oleuropein in isolated rat heart.

Previous studies have reported that oleuropein, the major constituent of olive leaves, has cardioprotective effects. There is no report related to oleuropein and ischemic-reperfusion injuries (cardiac dysfunction and myocardial infarction) as well as preconditioning in rat hearts. 56 male Wistar rats were divided into 7 groups (n=8). Group 1 as the control group and groups 2 to 7 as the treatment groups that received a single dose of oleuropein (100 mg/kg, i.p.) 1, 3, 6, 12, 24 and 48 hours before the excision of the heart, respectively. After these times, their hearts were excised and subjected to 30 min regional ischemia and 120 min reperfusion under Langendorff apparatus. Electrocardiogram and intraventricular pressures were monitored and recorded throughout the procedure. Finally, infarct size was measured by triphenyltetrazolium chloride staining. Compared to the control group, oleuropein significantly reduced infarct size and reperfusion-induced cardiac dysfunction in groups 2 and 3. Oleuropein markedly attenuated both ischemic and reperfusion arrhythmias in groups 2 and 3. There was no significant difference between other groups (4 to 7) than the control group. Heart rate had no significant difference among all of the groups. These results indicate that pretreatment of rats with a single dose of intraperitoneal oleuropein could protect their heart against ischemic-reperfusion injury for at least 3 hours. However, it has no preconditioning effect, since oleuropein had not cardioprotective effects 24 hour later.

Prolonged Oral Administration of Oleuropein Might Protect Heart against Aconitine-induced Arrhythmia.

In this study, it was surveyed to know whether an oral single dose of oleuropein could mimic the cardiac preconditioning in rats' hearts or whether its prolonged oral administration could protect the heart against the aconitine-induced arrhythmia in rats. Eighty male Wistar rats were divided into two series (n = 8 in each group). In the first series, all groups (except the control (Con) group) were given a single oral dose of oleuropein (20 mg/Kg) 1, 3, 24 and 48 h before the infusion of aconitine. In the second series, except the Con group, the other four groups were given oral oleuropein (20 mg/Kg/day) for 3, 7, 14 and 28 days, before the infusion of aconitine. Electrocardiogram was recorded to monitor arrhythmia. Data of the first series showed that the initiation time of arrhythmia, the initiation of ventricular tachycardia (VT), the numbers of reversible ventricular fibrillation (VF) and the death time had no significant difference compared with Con group. In the second series, a significant protection was occurred only in the 28 days group that was evident with increased initiation time of arrhythmia, increased initiation time of VT, and increased the number of reversible VF and death time in compared to the Con group. The findings of this study show that the oral administration of a single dose of oleuropein could not mimic the preconditioning effects in rat hearts, but the prolonged administration of oleuropein for about four weeks could protect the heart against aconitine-induced arrhythmia.

Anticonvulsant and depressant effects of aqueous extracts of Carum copticum seeds in male rats.

In this study, the effects of aqueous extracts of Carum copticum seeds (CCS) were evaluated in kindling models of epilepsy. Additionally, the sedative and anxiolytic effects of the extract were assessed. For pentylenetetrazole (PTZ) kindling, rats received a subconvulsant dose of PTZ (40 mg/kg, ip) every second day and seizure stages were recorded. CCS aqueous extract (200, 400, or 600 mg/kg, ip) was injected 30 minutes prior to each PTZ injection. In electrical kindling, bipolar stimulating and monopolar recording electrodes were implanted stereotaxically in the right basolateral amygdala of male Sprague-Dawley rats. After kindling, the effect of aqueous extracts of CCS (200, 400, or 600 mg/kg, ip) on afterdischarge duration, duration of rearing, forelimb clonus, and loss of equilibrium (stage 5 seizure), and latency to the onset of bilateral forelimb clonus were measured. The sedative and the anxiolytic effects of CCS extracts were evaluated in an open-field apparatus and elevated plus maze, respectively. The results indicate that aqueous extracts of CCS have a significant anticonvulsant effect. Different doses of extract significantly delayed the incidence of every seizure stage in the PTZ model of kindling. Moreover, CCS extract (400 and 600 mg/kg, ip) suppressed afterdischarge duration, latency to the onset of bilateral forelimb clonus, and stage 5 seizure in the electrical kindling model. These results suggest that CCS extract has remarkable antiepileptic and central depressant effects.

Pretreatment with hyperoxia reduces in vivo infarct size and cell death by apoptosis with an early and delayed phase of protection.

OBJECTIVE: Exposure to normobaric hyperoxia protects the heart against ischemia-reperfusion injury ex vivo. In the present study, we investigated the effect of the early and late phase of hyperoxia on in vivo myocardial infarction and apoptosis. METHODS: Rats were exposed to room air preoxygenation (O(2)≥ 95%) followed by regional ischemia (30 min) and 0, 90, 180, and 360 min of reperfusion. Hyperoxic exposure was performed for 120 min either immediately or 24h before coronary occlusion followed by 360-min reperfusion. Infarct size was evaluated by Evans blue/triphenyltetrazolium chloride staining. Apoptosis in the infarcted area was evaluated by terminal deoxy-nucleotidyl transferase-mediated deoxy uridine triphosphate (dUTP) nick end-labeling (TUNEL). Caspase 3 activity was measured by fluorometric enzyme assay, Bcl-2 and Bax protein expression assessed by western blotting and DNA laddering assessed with DNA gel electrophoresis. RESULTS: The infarct size did not increase with increasing duration of reperfusion. However, apoptosis as evaluated by Bcl-2/Bax ratio, caspase 3 activity, and TUNEL-positive cells increased with increasing time of reperfusion. Both early and delayed pretreatment with hyperoxia reduced infarct size (p = 0.0013, p = 0.046), ameliorated ischemic arrhythmias and increased Bcl-2/Bax ratio (p = 0.015, p = 0.0159). Only hyperoxia immediately before coronary occlusion decreased caspase 3 activity (p = 0.026) and decreased TUNEL-positive staining (p = 0.046) with no visible DNA laddering. CONCLUSIONS: Detection of myocardial apoptosis increased with prolongation of reperfusion time, as opposed to infarct detection where reperfusion was essential to detect infarction, but the infarct size did not increase with time. Pretreatment with hyperoxia significantly decreased infarct size and apoptotic cell death. Pretreatment, immediately before coronary occlusion, was most cardioprotective.

Ischemia and reperfusion-induced arrhythmias: role of hyperoxic preconditioning.

BACKGROUND: Hyperoxic preconditioning is known to protect the heart against necrosis and contractile dysfunction, but protection against arrhythmias has not been well characterized. OBJECTIVE: The authors hypothesized that pre-exposure to normobaric hyperoxia (H) reduces ischemia and reperfusion-induced arrhythmias in isolated rat hearts. METHODS: Following 60 and 180 min of hyperoxia treatment, rat hearts were isolated immediately (H60 and H180) or 24 h afterward (H60/24 and H180/24), and subjected to 30 min of regional ischemia followed by 120 min of reperfusion. Occurrence, number, and duration of arrhythmias were analyzed during ischemia and reperfusion. In addition, cardiac infarct size was also assessed. RESULTS: Sixty and 180 min of breathing hyperoxic gas induced significant protection against severe ischemia and reperfusion-induced arrhythmias. Total number of premature ventricular beats was markedly attenuated by hyperoxia pre-exposure, especially in H60 and H180 groups. Duration of ventricular tachycardia and ventricular fibrillation was also affected by hyperoxia. Hyperoxia reduced the number of ventricular tachycardia episodes in ischemia and reperfusion phase. Accordingly, severity of arrhythmias (arrhythmia score) and infarct size were lower in hyperoxia-treated groups. The effects were more pronounced using hyperoxia immediately before harvesting the heart. CONCLUSION: These results indicate that hyperoxic preconditioning attenuates ventricular ischemia and reperfusion-induced arrhythmias in isolated rat hearts, decreases cardiac infarct size, and improves postischemic heart function. The effects seem to depend on the time course after hyperoxia treatment.