Resveratrol and 1,25-dihydroxyvitamin D co-administration protects the heart against D-galactose-induced aging in rats: evaluation of serum and cardiac levels of klotho.

The current study investigates the cooperative cardioprotective effect of calcitriol (active form of vitamin D) combined with resveratrol in a rat model of D-galactose (D.gal)-induced aging. Male Wistar rats received resveratrol (D.gal + Res), calcitriol (D.gal + Cal), or a combination of them (D.gal + Res + Cal). Intact animals served as control (Ctl). Blood pressure (BP) was recorded by cannulation of the left carotid artery. Fibrosis and cell size were assessed by Masson's trichrome and hematoxylin-eosin staining, respectively. Cardiac and serum level of antiaging protein, klotho, was measured by ELISA assay method. Gene expression was evaluated by real-time RT-PCR. Biochemical tests were performed according to the standardized method. In D.gal + Res + Cal group, BP, heart weight-to-body weight ratio, and cardiomyocytes size decreased significantly compared with D-gal group. The cardiac transcription levels of catalase and superoxide dismutase 1 and 2 were upregulated in D.gal + Res + Cal compared to the D.gal group (P < 0.001, P < 0.05, P < 0.05, respectively). Increased level of malondialdehyde was observed in D.gal group (P < 0.01 vs. Ctl) which was normalized partially in D.gal + Res + Cal group (P < 0.05). Catalase and superoxide dismutase activity also increased in D.gal + Res + Cal group (P < 0.01 vs. D.gal). Cardiac Klotho, as the antiaging protein, remained unchanged at mRNA and protein levels among the experimental groups. The serum level of Klotho did not change significantly in D.gal group; however, in D.gal + Res + Cal group, serum klotho concentration was increased (P < 0.05 vs. D.gal). It could be concluded that co-administration of resveratrol and vitamin D protects the heart against aging-induced damage by the modulation of hemodynamic parameters and antioxidant status of the heart. Furthermore, increased serum level of klotho could be a novel mechanism for antiaging effects of resveratrol and vitamin D.

Effect of endogenous sulfur dioxide on spatial learning and memory and hippocampal damages in the experimental model of chronic cerebral hypoperfusion.

Background The vascular changes due to cerebrovascular damage, especially on the capillaries, play a vital role in causing vascular dementia. Increasing oxidative stress can lead to tissue damage while reducing brain blood flow. The use of factors reducing the oxidative stress level can decrease the brain damages. Sulfur dioxide (SO2) is one of the most important air pollutants that lead to the development of severe brain damage in large quantities. However, studies have recently confirmed the protective effect of SO2 in cardiac ischemic injury, atherosclerosis and pulmonary infections. Methods The permanent bilateral common carotid artery occlusion (BCAO) method was used to induce chronic cerebral hypoperfusion (CCH). Two treatment groups of SO2 were studied. The animal cognitive performance was evaluated using the Morris water maze. Hippocampal tissue damage was examined after 2 months of BCAO. In the biochemical analysis, the activity of catalase and lipid peroxidation of the hippocampus was studied. Results Neuronal damage in hippocampus, as well as cognitive impairment in ischemia groups treated with SO2 showed a significant improvement. Catalase activity was also significantly increased in the hippocampus of treated groups. Conclusions According to the results, SO2 is likely to be effective in reducing the CCH-caused damages by increasing the antioxidant capacity of the hippocampus.

Carvacrol suppresses learning and memory dysfunction and hippocampal damages caused by chronic cerebral hypoperfusion.

Chronic cerebral hypoperfusion (CCH) is a common phenomenon in many neurological diseases such as vascular dementia and Alzheimer's disease. Several drugs have been investigated to prevent and treat the CCH. The carvacrol (CAR) has been shown to have beneficial effects on various neurodegenerative and neuropsychiatric disorders. Accordingly, the present study was designed to evaluate the effect of CAR on neuronal damages in hippocampus in a well-defined model for CCH. Forty-eight male Wistar rats were equally divided into four groups of sham (A), CCH (B), CCH+ CAR 25, and 50 mg/kg/daily (C and D). The animals were subjected to permanent bilateral occlusion of the carotid arteries (2-vessel occlusion, 2VO) to induce CCH model. Cognitive function was evaluated by Morris water maze test. Morphological changes of hippocampus were assessed using Nissl staining. Free radical scavenging activity was measured by 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay. Moreover, catalase (CAT) and superoxide dismutase (SOD) activities and lipid peroxidation levels were measured using biochemical analysis. CAR significantly improved the spatial learning and memory deficits assessed using the Morris water maze test. CAR also significantly attenuated neuronal necrosis as well as malondialdehyde (MDA) and elevated the levels of SOD and CAT activity in the hippocampus. The results indicate that CAR produces significant neuroprotective effects on neuronal damages induced by CCH. Protective effect of CAR may be mediated by antioxidative effect of this drug.

Carvacrol Ameliorates Pathological Cardiac Hypertrophy in Both In-vivo and In-vitro Models.

Hypertension-induced left ventricular hypertrophy is the most important risk factor for heart failure. This study aimed at investigating the effects of monoterpenoid phenol, carvacrol, on myocardial hypertrophy using both in-vivo and in-vitro models. Male Wistar rats were divided into the control (Ctl), un-treated hypertrophy (H), and carvacrol-treated hypertrophy groups (25, 50 and 75 mg/kg/day, Car+H). In the hypertrophy groups animals underwent abdominal aorta banding. Blood pressure (BP) was recorded via carotid artery cannulation. TUNEL assay and Masson's trichrome staining were used to assess apoptosis and fibrosis, respectively. The 2-2-diphenyl 1-picril-hydrasil )DPPH( radical scavenging activity and malondialdehyde (MDA) level were estimated by biochemical tests. In in-vitro study H9c2 cardiomyoblasts were treated with angiotensin II (Ang II) to promote hypertrophy. Cell size was measured using crystal violet staining. Gene expression was evaluated by real-time RT-PCR technique. In the carvacrol-treated rats BP, heart rate, and heart weight to the body weight ratio were significantly decreased. In-vitro study showed that H9c2 cell size was significantly reduced compared to Ang II-treated cells. Both in-vivo and in-vitro studies demonstrated that carvacrol decreased atrial natriuretic peptide )ANP( mRNA level significantly (vs. H groups). The number of apoptotic cells increased in H group, while it was decreased in the Car50+H and Car75+H. In Car+H groups, in comparison with H group, the serum concentration of MDA was decreased and DPPH was increased significantly. Our findings demonstrated that carvacrol decreases hypertrophy markers in in-vivo and in-vitro models of hypertrophy.

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.

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.

Evaluation of Cytotoxicity Effects of Oleo-Gum-Resin and Its Essential Oil of Ferula assa-foetida and Ferulic Acid on 4T1 Breast Cancer Cells.

BACKGROUND: Cancer causes significant morbidity and mortality and is a major public health problem worldwide. Breast cancer is a leading cause of cancer-associated mortality in women, and the incidence is also on the rise in the entire world. Medicinal plants have been an important source of several clinically useful anticancer agents. AIM: In this study, we studied the growth inhibitory effect of asafoetida and its essential oil and ferulic acid on antitumor activity using mouse breast cancer cell line. MATERIALS AND METHODS: For this aim, cells were exposed to these components at different concentrations and for different time durations. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was carried out to characterize the cytotoxicity of the constituents used. RESULTS: Our results showed that all three constituents could inhibit 4T1 cell proliferation. Our MTT assay results showed a significant cytotoxicity effect in a time- and concentration-dependent manner. It also demonstrated that essential oil of asafoetida has a stronger effect in decreasing viability breast cancer cells. Ferulic acid showed a significant effect only at a dose of 500 µg/ml. CONCLUSIONS: Based on the results of cellular carried out in this study, we could demonstrate that asafoetida and its essential oil and ferulic acid have inhibitory effect on the growth of breast cancer cell line. As evidenced from these preliminary results, asafoetida and its derivative constituents may be considered as attractive alternatives to serve as lead compounds in drug development for breast cancer as an adjuvant therapy. However, much remains to be done before such agent could be introduced to the clinic.

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.