Mechanisms by Which Exercise Training Attenuates Blood Pressure in Animals: Roles of Cytokines, Oxidative Stress, and Glycogen Synthase Kinase-3â

Hypertension is a chronic multifactorial condition with high morbidity and mortality rates, currently affecting about one billion people worldwide. Currently available anti-hypertensive medications are found to be effective in reducing blood pressure (BP), but still more than 50% of those diagnosed with hypertension fail to respond to these anti-hypertensive regimens. Although hypertension has multiple etiologies, physical inactivity has been found to have strong correlation with the disease, so exercise has recently been recommended as a part of lifestyle modifications for all hypertensive patients. Therefore, the present series of in vivo and in vitro studies were undertaken to gain more insight into the effects of regular long-term exercise training (ExT) within the heart and brain of hypertensive animals with the specific aim of investigating the molecular mechanisms underlying the exercise-induced beneficial effects. In the first study, we subjected young spontaneously hypertensive rats (SHRs) to moderate-intensity exercise for 16 weeks. Regular exercise delayed progression of hypertension and improved cardiac function in SHRs, and these effects were mediated by reduced myocardial pro-inflammatory cytokines (PICs), NFêB activity, and improved redox homeostasis. In the second study, we found that chronic exercise not only reduces PICs and vasoconstrictor components of the renin-angiotensin system (RAS) but also improved anti-inflammatory cytokines (AIC) and vasodilatory axis of the RAS within the brain of SHRs. In the third study, we explored the effects of cessation of exercise (physical detraining) on these parameters. Next, we examined the role of GSK-3â in dysregulation of PICs and AIC in vitro using neuronal cell culture and in vivo using angiotensin II-induced hypertensive rat model. Finally, we investigated the effects of ExT on brain GSK-3â in hypertension and whether central GSK-3â mediates exercise-induced beneficial effects in hypertension. Collectively, these studies demonstrate that unlike pharmacological therapies, chronic regular exercise is a non-pharmacological cost-effective tool that has the capability to positively modulate several components of signaling pathways involved in pathogenesis of hypertension. These findings provide greater insight into the molecular mechanisms underlying the exercise-induced beneficial effects and will ultimately lead us to refine the current guidelines for the treatment of hypertension on the basis of scientific evidence

EFFECT OF Staphylococcus epidermidis ON Pseudomonas aeruginosa BIOFILM IN MIXED-SPECIES CULTURE

Staphylococcus epidermidis and Pseudomonas aeruginosa, are clinically relevant pathogens that often produce biofilms. To investigate the co-survivability of S. epidermidis and P. aeruginosa in mixed cultures biofilm and planktonic form, it is important to understand more about the interspecies interaction of both species. The interspecies interaction was analyzed using streak and drop agar plate assay, cell viability assay (CFU), spectrophotometry-based method, and microscopic analysis. The findings suggest that both cells and supernatant of P. aeruginosa inhibit the planktonic growth of S. epidermidis. The cell viability result shows that PAO1 biofilm cells were decreased by 88%, and SE biofilm cells were increased by 75% concerning their control. Opposite to the P. aeruginosa, the S. epidermidis biofilm and EPS matrix were found to increase in mixed culture biofilm, which was further confirmed by microscopic analysis. In contrast, differential agar media result shows that the reduction in the biofilm (CFU/ml) of P. aeruginosa is independent of S. epidermidis cells concentration. Finally, the effect of the supernatant on biofilm was investigated, and it found that S. epidermidis biofilm was enhanced while P. aeruginosa biofilm was reduced in the presence of partner bacterial supernatant, which indicated that S. epidermidis in biofilm mode could hinder the biofilm formation of P. aeruginosa. The outcomes show that the culture supernatant of S. epidermidis can be used to prevent P. aeruginosa associated biofilm infections

Toll-like receptor 4 inhibition within the paraventricular nucleus attenuates blood pressure and inflammatory response in a genetic model of hypertension

© 2015 Dange et al. Background: Despite the availability of several antihypertensive medications, the morbidity and mortality caused by hypertension is on the rise, suggesting the need for investigation of novel signaling pathways involved in its pathogenesis. Recent evidence suggests the role of toll-like receptor (TLR) 4 in various inflammatory diseases, including hypertension. The role of the brain in the initiation and progression of all forms of hypertension is well established, but the role of brain TLR4 in progression of hypertension has never been explored. Therefore, we investigated the role of TLR4 within the paraventricular nucleus (PVN; an important cardioregulatory center in the brain) in an animal model of human essential hypertension. We hypothesized that a TLR4 blockade within the PVN causes a reduction in mean arterial blood pressure (MAP), inflammatory cytokines and sympathetic drive in hypertensive animals. Methods: Spontaneously hypertensive rats (SHR) and normotensive Wistar Kyoto (WKY) rats were administered either a specific TLR4 blocker, viral inhibitory peptide (VIPER), or control peptide in their PVN for 14 days. MAP was recorded continuously by radiotelemetry. PVN and blood were collected for the measurement of pro-inflammatory cytokines (Tumor Necrosis Factor (TNF)-α, interleukin (IL)-1β), anti-inflammatory cytokine IL-10, inducible nitric oxide synthase (iNOS), TLR4, nuclear factor (NF) ΚB activity and plasma norepinephrine (NE) and high mobility group box (HMGB)1 expression, respectively. Results: Hypertensive rats exhibited significantly higher levels of TLR4 in the PVN. TLR4 inhibition within the PVN attenuated MAP, improved cardiac hypertrophy, reduced TNF-α, IL-1β, iNOS levels, and NFΚB activity in SHR but not in WKY rats. These results were associated with a reduction in plasma NE and HMGB1 levels and an increase in IL-10 levels in SHR. Conclusions: This study demonstrates that TLR4 upregulation in PVN plays an important role in hypertensive response. Our results provide mechanistic evidence that hypertensive response in SHR are mediated, at least in part, by TLR4 in the PVN and that inhibition of TLR4 within the PVN attenuates blood pressure and improves inflammation, possibly via reduction in sympathetic activity

Data regarding anti-quorum sensing and antimicrobial activity of Melaleuca alternifolia and Salvia sclarea essential oil against Pseudomonas aeruginosa

To combat the increasing number of multi-drug resistant bacteria, researchers are now looking for alternatives that reduce the virulence and pathogenic potential of the bacteria without killing it. It can be accomplished by interfering with the quorum sensing (QS) system of bacteria. In this article, we aim to determine the antimicrobial and anti-QS activity of Salvia sclarea and Melaleuca alternifolia essential oils (EOs) against Pseudomonas aeruginosa. The sub-lethal concentration of these EOs was found with the help of a growth curve, and further experiments were carried out below this concentration. To check for their anti-quorum activity, a bioreporter strain E. coli pJN105LpSC11 (to measure the concentration of 3-oxo-C12HSL) and Chromobacterium violaceum CV026 (to check for the reduction in the formation of violacein pigment) was used. Several virulence phenotype assays like pyocyanin, alginate, and protease production, along with swarming motility, were done. The effect of these EOs on biofilm formation was also checked. The results were confirmed by checking the expression of genes by real-time PCR

Angiotensin II causes imbalance between pro- and anti-inflammatory cytokines by modulating GSK-3β in neuronal culture

BACKGROUND AND PURPOSE: Emerging evidence indicates that the balance between pro-inflammatory cytokines (PICs) and anti-inflammatory cytokines (AICs) within the brain is an important determinant in the outcome of hypertension. However, the mechanism by which this dysregulation occurs is not known. We aimed to investigate whether AngII induces imbalance between PIC and AIC by modulating downstream transcription factors, NFκB and cyclic AMP response element-binding protein (CREB), and whether AngII-induced effects are mediated by glycogen synthase kinase-3β (GSK-3β). EXPERIMENTAL APPROACH: CATH.a neurons were exposed to AngII (10 nM-1 μM) over a preset time course. In another set of experiments, GSK-3β was knock down by using lentivirus containing short hairpin RNA targeting GSK-3β (L-sh-GSK3β) before AngII exposure. Cell extracts were subjected to RT-PCR, immunoblot and immunoprecipitation. KEY RESULTS: AngII caused time-dependent increase in PICs (TNF-α and IL-1β) and reduction in AIC (IL-10). AngII exposure caused reduced phosphorylated CREB(Ser-133) and increased p-NFκB(Ser-276) levels, leading to reduced CREB-CBP and increased NFκB-CBP binding. These results were accompanied by increased activation of GSK-3β, as indicated by increased p-GSK3(Tyr-216) to p-GSK3(Ser-9) ratio. In a subsequent study, pretreatment with L-sh-GSK3β attenuated AngII-induced alterations in PICs and IL-10 by augmenting CREB-CBP and attenuating NFκB-CBP binding. CONCLUSIONS AND IMPLICATIONS: Collectively, these findings are the first to provide direct evidence that AngII-induced dysregulation in cytokines is mediated by GSK-3β-mediated alterations in downstream transcription factors in neuronal cells. Our data also reveal that AngII-induced effects could be alleviated by GSK-3β inhibition, suggesting GSK-3β as an important therapeutic target for hypertension that is characterized by increased PICs and NFκB activation

Angiotensin II-induced hypertension is modulated by nuclear factor-κBin the paraventricular nucleus

Hypertension is considered a low-grade inflammatory condition, and understanding the role of transcription factors in guiding this response is pertinent. A prominent transcription factor that governs inflammatory responses and has become a focal point in hypertensive research is nuclear factor-κB (NFκB). Within the hypothalamic paraventricular nucleus (PVN), a known brain cardioregulatory center, NFκB becomes potentially even more important in ultimately coordinating the systemic hypertensive response. To definitively demonstrate the role of NFκB in the neurogenic hypertensive response, we hypothesized that PVN NFκB blockade would attenuate angiotensin II-induced hypertension. Twelve-week-old male Sprague-Dawley rats were implanted with radiotelemetry probes for blood pressure measurement and allowed a 7-day recovery. After baseline blood pressure recordings, rats were administered either continuous NFκB decoy oligodeoxynucleotide infusion or microinjection of a serine mutated adenoviral inhibitory-κB vector, or their respective controls, bilaterally into the PVN to inhibit NFκB at two levels of its activation pathway. Simultaneously, rats were implanted subcutaneously with an angiotensin II or saline-filled 14-day osmotic minipump. After the 2-week treatments, rats were euthanized and brain tissues collected for PVN analysis. Bilaterally inhibited NFκB rats had a decrease in blood pressure, NFκB p65 subunit activity, proinflammatory cytokines, and reactive oxygen species, including the angiotensin II type 1 receptor, angiotensin-converting enzyme, tumor necrosis factor, and superoxide in angiotensin II-treated rats. Moreover, after NFκB blockade, key protective antihypertensive renin-angiotensin system components were upregulated. This demonstrates the important role that transcription factor NFκB plays within the PVN in modulating and perpetuating the hypertensive response via renin-angiotensin system modulation

Chronic exercise modulates RAS components and improves balance between pro- and anti-inflammatory cytokines in the brain of SHR

Recently, exercise has been recommended as a part of lifestyle modification for all hypertensive patients; however, the precise mechanisms of its effects on hypertension are largely unknown. Therefore, this study aimed to investigate the mechanisms within the brain that can influence exercise-induced effects in an animal model of human essential hypertension. Young normotensive WKY rats and SHR were given moderate-intensity exercise for 16 weeks. Blood pressure was measured bi-weekly by tail-cuff method. Animals were then euthanized; paraventricular nucleus (PVN) and rostral ventrolateral medulla (RVLM), important cardiovascular regulatory centers in the brain, were collected and analyzed by real-time RT-PCR, Western blot, EIA, and fluorescent microscopy. Exercise of 16-week duration attenuated systolic, diastolic, and mean arterial pressure in SHR. Sedentary SHR exhibited increased pro-inflammatory cytokines (PICs) and decreased anti-inflammatory IL-10 levels in the PVN and RVLM. Furthermore, SHR(sed) rats exhibited elevated levels of ACE, AT1R, and decreased levels of ACE2 and receptor Mas in the PVN and RVLM. Chronic exercise not only prevented the increase in PICs (TNF-α, IL-1β), ACE, and AT1R protein expression in the brain of SHR, but also dramatically upregulated IL-10, ACE2, and Mas receptor expression in SHR. In addition, these changes were associated with reduced plasma AngII levels, reduced neuronal activity, reduced NADPH-oxidase subunit gp91(phox) and inducible NO synthase in trained SHRs indicating reduced oxidative stress. These results suggest that chronic exercise not only attenuates PICs and the vasoconstrictor axis of the RAS but also improves the anti-inflammatory defense mechanisms and vasoprotective axis of the RAS in the brain, which, at least in part, explains the blood pressure-lowering effects of exercise in hypertension

Detraining differentially preserved beneficial effects of exercise on hypertension: effects on blood pressure, cardiac function, brain inflammatory cytokines and oxidative stress.

AIMS: This study sought to investigate the effects of physical detraining on blood pressure (BP) and cardiac morphology and function in hypertension, and on pro- and anti-inflammatory cytokines (PICs and AIC) and oxidative stress within the brain of hypertensive rats. METHODS AND RESULTS: Hypertension was induced in male Sprague-Dawley rats by delivering AngiotensinII for 42 days using implanted osmotic minipumps. Rats were randomized into sedentary, trained, and detrained groups. Trained rats underwent moderate-intensity exercise (ExT) for 42 days, whereas, detrained groups underwent 28 days of exercise followed by 14 days of detraining. BP and cardiac function were evaluated by radio-telemetry and echocardiography, respectively. At the end, the paraventricular nucleus (PVN) was analyzed by Real-time RT-PCR and Western blot. ExT in AngII-infused rats caused delayed progression of hypertension, reduced cardiac hypertrophy, and improved diastolic function. These results were associated with significantly reduced PICs, increased AIC (interleukin (IL)-10), and attenuated oxidative stress in the PVN. Detraining did not abolish the exercise-induced attenuation in MAP in hypertensive rats; however, detraining failed to completely preserve exercise-mediated improvement in cardiac hypertrophy and function. Additionally, detraining did not reverse exercise-induced improvement in PICs in the PVN of hypertensive rats; however, the improvements in IL-10 were abolished. CONCLUSION: These results indicate that although 2 weeks of detraining is not long enough to completely abolish the beneficial effects of regular exercise, continuing cessation of exercise may lead to detrimental effects

Role of proinflammatory cytokines and redox homeostasis in exercise-induced delayed progression of hypertension in spontaneously hypertensive rats

Hypertension is a well-known risk factor for various cardiovascular diseases. Recently, exercise has been recommended as a part of lifestyle modification for all hypertensive patients. However, the precise mechanisms of exercise training (ExT)-induced effects on the development of hypertension are poorly understood. Therefore, we hypothesized that chronic ExT would delay the progression of hypertension in young spontaneously hypertensive rats (SHRs). In addition, we explored whether the beneficial effects of chronic ExT were mediated by reduced proinflammatory cytokines and improved redox status. We also investigated the involvement of nuclear factor-kappaB in exercise-induced effects. To test our hypotheses, young normotensive (Wistar-Kyoto) and SHRs were given moderate-intensity ExT for 16 weeks. Blood pressure was determined by the tail-cuff method, and cardiac function was assessed by echocardiography. Myocardial total reactive oxygen species and superoxide production were measured by electron paramagnetic resonance spectroscopy; tumor necrosis factor-alpha, interleukin-1beta, gp91(phox), and inducible NO synthase by real-time PCR; and nuclear factor kappaB activity by electrophoretic mobility shift assay. Chronic ExT in hypertensive rats resulted in significantly reduced blood pressure, reduced concentric hypertrophy, and improved diastolic function. ExT significantly reduced proinflammatory cytokines and inducible NO synthase, attenuated total reactive oxygen species and superoxide production, and increased antioxidants in SHRs. ExT also resulted in increased NO production and decreased nuclear factor kappaB activity in SHRs. In summary, chronic ExT delays the progression of hypertension and improves cardiac function in young SHRs; these ExT-induced beneficial effects are mediated by reduced proinflammatory cytokines and improved redox homeostasis via downregulation of nuclear factor-kappaB