The Combination Neuroprotective Abilities of Resveratrol and Naringenin in Attenuation of Sleep Deprivation Complications in Rats

Background: Sleep loss is one of the most important health problems in the world, and about 30 to 40 percent of ordinary people suffer from it. This study aimed to investigate the neuroprotective effects of the combination of resveratrol and naringenin in attenuation of sleep deprivation (SD) complications in rats. Materials and Methods: In this experimental study, 72 Wistar male rats were randomly divided into three main groups, including control, sham, and 7-days SD group. Each of its main groups consisted of three subgroups, including without drug, vehicle, and combination therapy groups (naringenin [100 mg/kg], resveratrol [100 mg/kg]). The day after the latest injection, the fear conditioning memory tests, locomotor activity test, hot plate, and forced swimming tests (FST) were carried out on all rats, and then sham and SD groups were induced 48 hours of non-REM SD (device off and on, respectively) and these behavioral tests were repeated for all rats again. Finally, the brains of all rats were removed and histopathologically examined, and stained with nissl and TUNNEL. Results: To assess fear condition memory, the rate of latency to first freezing in the visual and auditory phase increased in sham and SD rats that received vehicle or no drug (P<0.001), which indicates memory corruption. Injection of the combination of naringenin and resveratrol reduced the latency to first freezing (P<0.001), which means improved memory. In the FST test, injection of naringenin and resveratrol reduced the rate of immobility (P<0.001), which means improved depressive behavior. The naringenin and resveratrol reduced the pain perception threshold. Also, the naringenin and resveratrol reduced apoptosis compared to the control and vehicle groups (P<0.001). Conclusions: The combination of naringenin and resveratrol compared to other groups could improve memory and mood as well as reduce apoptosis, depression, and pain perception threshold. [GMJ.2021;10:e2315]

Modulation of neural activity in frontopolar cortex drives reward-based motor learning

The frontopolar cortex (FPC) contributes to tracking the reward of alternative choices during decision making, as well as their reliability. Whether this FPC function extends to reward gradients associated with continuous movements during motor learning remains unknown. We used anodal transcranial direct current stimulation (tDCS) over the right FPC to investigate its role in reward-based motor learning. Nineteen healthy human participants practiced novel sequences of finger movements on a digital piano with corresponding auditory feedback. Their aim was to use trialwise reward feedback to discover a hidden performance goal along a continuous dimension: timing. We additionally modulated the contralateral motor cortex (left M1) activity, and included a control sham stimulation. Right FPC-tDCS led to faster learning compared to lM1-tDCS and sham through regulation of motor variability. Bayesian computational modelling revealed that in all stimulation protocols, an increase in the trialwise expectation of reward was followed by greater exploitation, as shown previously. Yet, this association was weaker in lM1-tDCS suggesting a less efficient learning strategy. The effects of frontopolar stimulation were dissociated from those induced by lM1-tDCS and sham, as motor exploration was more sensitive to inferred changes in the reward tendency (volatility). The findings suggest that rFPC-tDCS increases the sensitivity of motor exploration to updates in reward volatility, accelerating reward-based motor learning.

The cross-talk between matrix metalloproteinase-9, RANKL/OPG system and cardiovascular risk factors in ovariectomized rat model of postmenopausal osteoporosis

Epidemiology and pathogenesis of cardiovascular diseases (CVD) and osteoporosis are strikingly overlapping. This study presents matrix metalloproteinase-9 (MMP-9), as a simple molecular link more consistently associated with the pathophysiology of both osteoporosis and CVD risk factors. 40 adult female rats were randomly distributed into 4 groups [control sham-operated, untreated osteoporosis, carvedilol-treated osteoporosis and alendronate-treated osteoporosis]. After 8 weeks, blood samples were collected to estimate Lipid profile (Total cholesterol, HDL, Triglycerides), inflammatory markers (IL-6, TNF alpha, CRP and NO), and Bone turnover markers (BTM) (Alkaline phosphatase, osteocalcin and pyridinoline). The tibias were dissected to estimate MMP-9 and NF-kB gene expression, OPG, RANKL levels and for histological examination. Induction of osteoporosis resulted in a significant elevation in BTM, inflammatory markers and dyslipidemia. MMP-9 was significantly elevated and positively correlated with BTM, inflammation and dyslipidemia markers. Carvedilol and alendronate exerted a bone preservative role and attenuated dyslipidaemia and inflammation in accordance with their respective effect on MMP-9.

Apigenin Acts as Anti-inflammatory Compound through Reducing IL-1β, IL-6 and TNF-α in LPS-induced Inflammation; in-vivo Study

Background: Consumption of herbal flavonoids instead of chemical drugs has increased in recent years due to fewer side effects and affordability. In this study, the effect of apigenin was investigated on inflammation induced by lipopolysaccharide in male rat's serum by measuring the pro-inflammatory cytokines, i.e., IL-1β, IL-6, and TNF-α.Methods: 90 male Wistar rats weighing 200 ±2 grams were used and divided into control, sham (solvent), and positive control (dexamethasone 15 mg/kg. ip), and 3 experimental groups which received 5, 15 or 30 mg/kg of apigenin, intraperitoneally. In 30 minutes after interventions, lipopolysaccharide (LPS) [30 μg/kg. ip] was injected. Then, at 4, 12- and 24-hour intervals, rats were anesthetized, and blood samples were prepared intracardially. Samples were centrifuged, and serums were separated and stored at -80 ° C. Measurement of IL-1β, IL-6, and TNF-α were conducted by the enzyme-linked immunosorbent assay (ELISA) method. Data were analyzed using the SPSS software version 19.Results: Pre-injection of apigenin at 5 mg/kg dosage were reduced TNF-α and IL-1β levels at 24-hours after LPS injection, compared to control (for both P <0.05). Pre-injection of 15 mg/kg of apigenin was reduced IL-6 level at 24-hours after LPS injection (P <0.05). Pre-injection of 30 mg/kg of apigenin were reduced TNF-α level at 4- (P <0.05), 12- (P <0.01) and 24- (P <0.01) hours, IL-1β level at 24-hours (P <0.01), and IL-6 level at 4- (P <0.05) and 24- (P <0.01) hours after LPS injection.Conclusions: Apigenin reduces proinflammatory cytokines in serum in acute inflammation induction. This impact is close to the dexamethasone effect as an anti-inflammatory steroid drug.

The Effects of Intermittent Hypoxic–Hyperoxic Exposures on Lipid Profile and Inflammation in Patients With Metabolic Syndrome

Background: Patients with metabolic syndrome (MS) tend to suffer from comorbidities, and are often simultaneously affected by obesity, dysglycemia, hypertension, and dyslipidemia. This syndrome can be reversed if it is timely diagnosed and treated with a combination of risk factors-reducing lifestyle changes and a tailored pharmacological plan. Interval hypoxic-hyperoxic training (IHHT) has been shown as an effective program in reducing cardiovascular risk factors in patients with MS even in the absence of exercise. However, the influence of IHHT on the lipid profile and inflammation in this clinical population remains relatively unknown.Methods: A prospective, single-center, randomized controlled trial was conducted on 65 (33 men) patients with MS aged 29–74 years, who were randomly allocated to the IHHT or control (sham) experimental groups. The IHHT group completed a 3-week, 5 days/week intermittent exposure to hypoxia and hyperoxia. The control (sham) group followed the same protocol but was breathing room air instead. The primary endpoints were the lipid profile (concentrations of total cholesterol [TC], low-density lipoprotein [LDL], high-density lipoprotein [HDL], and triglycerides [TG]) and the inflammatory factors such as high-sensitivity C-reactive protein (hs-CRP), galectin-3, heat shock proteins (Hsp70). The secondary endpoints were alanine aminotransferase (ALT), aspartate aminotransferase (AST), N-terminal pro-hormone of brain natriuretic peptide level (NTproBNP), transforming growth factor beta-1 (TGF-beta1), heart-type fatty acid-binding protein (H-FABP), and nitric oxide synthase 2 (NOS2).Results: There were no differences between the two groups but the different baseline values have affected these results. The IHHT group demonstrated pre-post decrease in total cholesterol (p = 0.001), LDL (p = 0.001), and TG levels (p = 0.001). We have also found a decrease in the CRP-hs (p = 0.015) and Hsp70 (p = 0.006) in IHHT-group after intervention, and a significant decrease in pre-post (delta) differences of NTproBNP (p &lt; 0.0001) in the IHHT group compared to the control group. In addition, the patients of the IHHT group showed a statistically significant decrease in pre-post differences of ALT and AST levels in comparison with the control group (p = 0.001). No significant IHHT complications or serious adverse events were observed.Conclusions: The IHHT appears to improve lipid profile and anti-inflammatory status. It is a safe, well-tolerated procedure, and could be recommended as an auxiliary treatment in patients suffering from MS, however, the experiment results were limited by the baseline group differences.Clinical Trial Registration:ClinicalTrials.gov, identifier [NCT04791397]. Evaluation of the effect of IHHT on vascular stiffness and elasticity of the liver tissue in patients with MS.

Comparing the Therapeutic Effects of Crocin, Escitalopram and Co-Administration of Escitalopram and Crocin on Learning and Memory in Rats with Stress-Induced Depression

Background: Depression affects various brain functions. According to previous studies, escitalopram influences brain functions in depression and crocin reduces memory impairments. Therefore, this study aimed to compare the therapeutic effects of using crocin and escitalopram (separately and in combination) on learning and memory in rats with stress-induced depression. Methods: Fifty-six rats were allocated into seven groups of control, sham, continuous depression, recovery period, daily injections of escitalopram, crocin and escitalopram-crocin during 14 days after inducing depression by stress. Passive avoidance (PA) test was used to assess brain functions. Results: Latency has significant differences in depression group. Also, it significantly increased in depression-crocin, depression-escitalopram and depression-escitalopram-crocin groups compared to the depression group. The dark stay (DS) time was significantly higher in the depression and depression-recovery groups. However, the DS time significantly decreased in the depression-crocin, depression-escitalopram and depression-escitalopram-crocin groups. Furthermore, the number of entrances to the dark room was significantly lower in depression-crocin and depression-escitalopram-crocin groups compared to the depression one. Conclusion: Different depression treatments (i.e. crocin, escitalopram and crocin- escitalopram) reduced depression-induced memory deficits. Crocin and escitalopram-crocin, respectively, improved brain functions and locomotor activity more than escitalopram. Comparatively, in subjects with depression, crocin, which is an effective saffron constituent, partially affected the memory deficits better than escitalopram (as a chemical component).

Neuroregenerative Effects of Intraspinal Microstimulation (ISMS) Following Spinal Cord Injury (SCI)

Background Intraspinal microstimulation (ISMS) is a novel electrical stimulation technique that has demonstrated mobility restoration in animals with spinal cord injury (SCI). This project investigated: 1) the capacity of ISMS to restore functional walking in rats with SCI through 4 weeks of stimulation, and 2) the degree of walking deficit caused by ISMS surgery. Methods Thirteen Sprague Dawley rats were divided into three groups: 1) rats with hemi-section SCI (hSCI) and no implants (control group), 2) rats with hSCI and passive ISMS implants (ISMS sham group), and 3) rats with hSCI and implants with active electrical stimulation (ISMS group). All groups were trained to walk on a horizontal ladder and their performance was quantified pre- and post-surgery. Results We hypothesized that the rats with active ISMS implants would demonstrate the greatest improvement in functional walking compared to both control groups, and that the ISMS sham group would underperform the most. The preoperative functional walking scores of control, sham and ISMS rats were 5.7±0.2, 5.5±0.3 and 5.7±0.1, respectively (7-point scale; mean ± standard error). The post-surgery scores were 3.2±0.9, 2.6±0.6 and 3.3±0.8 for control, sham, and ISMS rats, respectively. Conclusions As the difference between the post-surgery functional walking scores of ISMS and control rats was not statistically significant, this may indicate that four weeks of ISMS stimulation is not enough to cause rehabilitative effects. Additionally, the ISMS sham group demonstrated impaired functional walking compared to the hSCI control group as predicted. Future studies will employ a larger sample size to fully elucidate this trend and utilize thinner microwires to mitigate cellular damage.

Local Anesthetic Delivered with a Dual Action Ring and Injection Applicator Reduces the Acute Pain Response of Lambs during Tail Docking

Docking the tail of lambs is a standard husbandry procedure and is achieved through several techniques including clamps, hot or cold knives and latex rings, the last of which is the most popular. All tail docking methods cause acute pain which can be reduced by application of local anesthetic, however precise anatomical injection for optimal efficacy requires considerable skill. This pen trial evaluated the ability of local anesthetic (LA) delivered with a dual function ring applicator/injector to alleviate acute tail docking pain. Thirty ewe lambs were assigned to one of three treatment groups (n = 10 per group): ring plus local anesthetic (Ring LA), ring only (Ring) and sham handled control (Sham). Lambs were videoed and their behavior categorized every five minutes for the first hour and every 10 min for the subsequent two hours after treatment. There was a significant effect (p < 0.001) of treatment on total active pain related behaviors in the first hour, with Ring lambs showing higher counts compared to Ring LA or Sham. Ring lambs also displayed a significantly higher count of combined abnormal postures (p < 0.001) than Ring LA or Sham lambs. Delivery of 1.5 mL of 2% lignocaine via the dual action device abolished abnormal behaviors and signs of pain in Ring LA lambs. However, lambs in the Ring LA group spent less time attempting to suckle compared to Ring and Sham lambs, suggesting that some residual discomfort remained.

Combination effects of gallic acid and cyclosporine A during ischemia/ reperfusion on rat electrocardiogram parameters and arrhythmia

Introduction: Myocardial ischemia leads to electrical disturbance in the heart because of reactive oxygen specious. This study aimed to investigate the effects of gallic acid and cyclosporine A (CsA) together on electrocardiogram parameters in myocardium following ischemia - reperfusion (I/R) in isolated hearts. Methods: In this research, 50 Wistar rats weighing 250-300g were randomly divided into the 5 following groups: control, sham and gallic acid (7.5, 15 and 30mg/kg) in combination with CsA (0.2μM). On the eleventh day, the hearts were removed and perfused with Krebs solution and ischemia was induced for 30min. Then, cyclosporine was administered for 10min at the 10 minutes before reperfusion and 10 minutes the beginning of reperfusion. By placing the electrode, the parameters of RR, PR, QT, TpeakTend, JT and QTcB interval, ST elevation, R, P, Q, S, T amplitude were recorded before ischemia and during reperfusion. Results: This study showed that RR, JT, interval, p duration, ST elevation and PVC numbers of control were increased during ischemia compared with sham and decreased using gallic acid (7.5, 15 and 30mg/kg) in combination with CsA. In addition, P, R, S, T amplitude during the ischemia were decreased in control compared with sham and increased with gallic acid (15mg/kg) in combination with CsA. Conclusion: In conclusion, the optimal combination of both drugs decreased arrhythmia occurrence while increased electrical velocity of conduction and wave amplitudes in isolated myocardium after ischemia reperfusion injury.

Decreased Substrate Stiffness Promotes a Hypofibrotic Phenotype in Cardiac Fibroblasts

A hypofibrotic phenotype has been observed in cardiac fibroblasts (CFs) isolated from a volume overload heart failure model, aortocaval fistula (ACF). This paradoxical phenotype results in decreased ECM synthesis despite increased TGF-β presence. Since ACF results in decreased tissue stiffness relative to control (sham) hearts, this study investigates whether the effects of substrate stiffness could account for the observed hypofibrotic phenotype in CFs isolated from ACF. CFs isolated from ACF and sham hearts were plated on polyacrylamide gels of a range of stiffness (2 kPa to 50 kPa). Markers related to cytoskeletal and fibrotic proteins were measured. Aspects of the hypofibrotic phenotype observed in ACF CFs were recapitulated by sham CFs on soft substrates. For instance, sham CFs on the softest gels compared to ACF CFs on the stiffest gels results in similar CTGF (0.80 vs. 0.76) and transgelin (0.44 vs. 0.57) mRNA expression. The changes due to stiffness may be explained by the observed decreased nuclear translocation of transcriptional regulators, MRTF-A and YAP. ACF CFs appear to have a mechanical memory of a softer environment, supported by a hypofibrotic phenotype overall compared to sham with less YAP detected in the nucleus, and less CTGF and transgelin on all stiffnesses.