Chronic Rose Oxide and Exercise Synergistically Modulate Cardiovascular and Autonomic Functions in Hypertensive Rats

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P. Araújo, Ana Flávia M. da Silva, and 14 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4939277/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 30 Oct, 2024 Read the published version in Pflügers Archiv - European Journal of Physiology → Version 1 posted 10 You are reading this latest preprint version Abstract With the alarming rise in cases of arterial hypertension worldwide, there is an urgent need to develop combined therapies to mitigate this scenario. Rose Oxide (RO), a monoterpene with anti-inflammatory and hypotensive properties, emerges as an alternative. The present study is the first to evaluate the effect of RO administered chronically and combined with physical exercise (swimming) since both have been reported to have beneficial impacts on hypertension. Male SHR and Wistar rats (aged 12 weeks) received RO for 34 consecutive days (orally; 100 mg/kg). The progression of systolic blood pressure (SBP) was monitored through tail-cuff plethysmography. Twenty-four hours before the end of the treatment, the animals were anesthetized, and the femoral artery and vein were cannulated to record the pulsatile arterial pressure and to administer drugs, respectively. Hemodynamic and autonomic parameters and baroreflex sensitivity and intrinsic heart rate (IHR) were evaluated. Treatment with RO, administered alone or combined with exercise, reduced SBP and mean arterial pressure in SHR. The swimming protocol did not prevent increases in BP, but when combined with RO, it improved autonomic control, assessed through heart rate variability and parasympathetic tone. IHR was attenuated in SHR, and none of the treatments reversed this response. Therefore, combining RO with physical exercise may enhance their antihypertensive effects, improving autonomic function, reducing oxidative stress and inflammation, providing synergistic cardiovascular benefits, improving metabolic health, promoting a comprehensive lifestyle intervention, and potentially allowing for reduced medication dosages. This multifaceted approach could offer a more effective and sustainable strategy for managing hypertension. Autonomic Control Rose Oxide Hypertension SHR Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 1 Introduction Analyzing the global context in recent years, there has been a noticeable increase in the number of people affected by some type of cardiovascular disease (CVD). In this alarming scenario, arterial hypertension (AH) is among the most prevalent CVD [ 1 ]. It is estimated that approximately one-third of the world population has blood pressure levels ≥ 140/90 mmHg, characterizing AH, whose treatment costs are high and mortality rate has risen by 56.1% in recent years [ 2 – 5 ]. Furthermore, unbalances in cardiac autonomic function, such as arterial baroreflex dysfunction and severe sympathetic hyperactivity combined with a decrease in parasympathetic activity, which characterizes a sympathetic-vagal imbalance, are known to exist in the pathogenesis of AH [ 6 – 8 ]. In this context, the evaluation of markers of cardiac autonomic control, such as baroreflex and parasympathetic tone, is of utmost importance for the monitoring and control of this disorder. In this respect, Heart Rate Variability (HRV) is a widely used mathematical model to infer autonomic activity and it is used in studies with both humans and animals [ 9 – 11 ]. Interestingly, the secondary metabolites called terpenes are promising therapeutic molecules in controlling AH [ 12 – 13 ]. A subdivision of terpenes called monoterpenes has gained prominence due to their vast diversity, abundance, and pharmacological properties in the circulatory system [ 14 ] In this perspective, Rose Oxide (RO), a tetrahydropyran of the monoterpene class, has shown therapeutic potential in the treatment of AH, considering its anti-inflammatory and antihypertensive actions [ 9 , 15 ] . On the other hand, besides pharmacological interventions, both resistance exercise and aerobic physical exercise (PE) are characterized as an excellent alternative approach capable of delaying AH development and enhancing its treatment, which potentially reduces mortality rates in some clinical outcomes. Moreover, it is known that PE promotes numerous physiological adaptations, such as improved endothelial function, glucose uptake, parasympathetic autonomic activity, and angiogenesis, neutralizes the generation of reactive oxygen species and promotes cardiac hypertrophy, among other benefits [ 16 – 18 ]. Thus, it is believed that combining pharmacological intervention with PE in the treatment of AH could be an effective therapeutic strategy for controlling this disease, thereby promoting better life expectancy, and reducing the amount or dosage of medications, directly influencing efficacy [ 19 ]. Therefore, our objective was to evaluate the chronic hypotensive activity of RO in combination with PE (swimming exercise). For this purpose, we used an inbred genetic model of experimental hypertension (Spontaneously Hypertensive Rats, SHR) to mimic AH that is observed in humans [ 9 , 20 ] 2 Methodology 2.1. Animals Experiments were conducted with Wistar rats and SHR provided by the Federal University of Piauí (UFPI). For the criteria of animal use and environmental control, we followed the same protocol proposed by Santos et al. [ 9 ]. It is worth noting that all experimental procedures adhered to the guidelines of the National Institutes of Health for the care and use of laboratory animals and were approved by the Ethics Committee on Animal Use of the Federal University of Piauí (CEUA/UFPI # 563/19). 2.2. Experimental Groups The animals were divided into five groups, as follows: 1) Wistar – control; 2) SHR – control; 3) RO SHR – administration of RO; 4) TSHR – aerobic training intervention; 5) RO TSHR – a combination of aerobic training and RO administration. 2.3. Reagents (+)-rose oxide (tetrahydro-4-methyl-2-(2-methylprop-1-enyl) pyran) was acquired from Sigma-Aldrich (Barueri, SP, Brazil; 99.0% purity), diluted in 2% Tween 20 P.S. (Vetec Química Fina Ltda.) and saline (0.9% sodium chloride). The dose used to evaluate the effect of RO was 100 mg/kg (adapted from Maia et al., 2021 [ 21 ]; Nonato et al., 2012 [ 15 ]). During the pulsatile arterial pressure (PAP) monitoring, the following drugs were used: phenylephrine (4 µg/kg i.v.), sodium nitroprusside [NPS (8 µg/kg i.v.)], methyl-atropine (2 mg/kg, i.v.), and propranolol (4 mg/kg, i.v.), all acquired from Sigma-Aldrich, St. Louis, MO, USA. For animal euthanasia, sodium thiopental (Cristália Prod. Quím. Farm. Ltda; injectable solution; 100 mg/kg; i.v.) was used at the end of the PAP recording, as established by Resolution No. 1,000 of May 11, 2012, of the Brazilian Federal Council of Veterinary Medicine – CFMV. 2.4. Tail-cuff plethysmography (TCP) To monitor the evolution of SBP in the animals during treatment, blood pressure (BP) monitoring was conducted using tail-cuff plethysmography (ADInstruments, Australia), following the protocol described by Silva et al. [ 22 ]. Plethysmography was performed at 4 time points: day 0 (baseline), week 1, week 3, and week 5 of treatment with RO or vehicle. 2.5. Gavage RO or vehicle was administered orally using a gavage needle for 34 consecutive days, according to the protocol by Machholz et al. [ 23 ]. 2.6. Swimming training Aerobic swimming training protocol, adapted from Rocha [ 24 ], was employed. All training groups initially underwent an adaptation period, which involved swimming in a cylindrical tank (80.0 cm in diameter, 120 cm deep) filled with thermo-neutral water (temperature of 30 ± 1°C) at a depth of approximately 100 cm. To minimize animal stress, the free-swimming time interval gradually increased as follows: 1st day − 15 min; 2nd day − 30 min; 3rd day − 45 min; and 4th day − 60 min. Subsequently, the rats swam for 4 weeks, swimming for 60 minutes per day, 5 times a week. 2.7. Surgical procedure for femoral artery and vein cannulation During the surgical procedure, the anesthetics ketamine (33.33 mg/kg) and xylazine (13.3 mg/kg) were used. The vessel cannulation procedure was performed as Santos et al. [ 9 ] and Oliveira [ 25 ] described. 2.8. Pulsatile arterial pressure monitoring During the PAP recording, the arterial catheter was connected to a pressure transducer (MLT0380/D, ADInstruments, Sydney, Australia). The signal, after proper amplification (ML110 Amplifier, ADInstruments, Sydney, Australia), was digitized through an analog-to-digital interface (ML866, ADInstruments, Sydney, Australia) and displayed at 2000 Hz on a microcomputer. Additionally, from the PAP recording, it was possible to extract mean arterial pressure (MAP) and heart rate (HR). The venous cannula was connected to a PE tube extension to facilitate drug administration, aiming to minimize animal stress during monitoring. Monitoring was divided into three parts: a period of animal acclimatization in the system (60 minutes), followed by baseline blood pressure recording (30 minutes), and concluding with the administration of vasoactive compounds. 2.9. Heart rate and systolic arterial pressure (SAP) variability Analysis of pulse interval (PI) and SAP variability was conducted considering both time-domain (TD) and frequency-domain (FD) parameters using CardioSeries software (v2.7- CardioSeries Software - CardioSeries Software (danielpenteado.com) ). To collect data, segments were extracted from the baseline period of pulsatile arterial pressure (PAP) monitoring. In the time domain, SAP variability was quantified using mean and standard deviation (SD), while PI was analyzed using mean and the square root of the mean of the sum of the squares of the differences between successive PI values (RMSSD). Regarding spectral analysis in the frequency domain, time series of SAP and PI were interpolated at 10 Hz and divided into continuous segments of 512 beats with 50% overlap. Each segment underwent Hanning windowing, and spectral analysis was performed using the Fast Fourier Transform (FFT). Oscillatory components were quantified in low-frequency (LF: 0.2–0.75 Hz) and high-frequency (HF: 0.75–3 Hz) bands. To assess sympathovagal balance, the LF/HF ratio was also calculated. Furthermore, the relative (%) and absolute power of the spectra within each frequency band were analyzed (Santos et al. 2021 [ 9 ] protocol; Sabino et al. 2013 [ 26 ]). 2.10. Statistical analysis The results are expressed as mean ± SEM (Standard Error of the Mean). Statistical analyses were performed using SigmaPlot software (version 11.0). Baseline hemodynamic values and the alterations induced by RO and/or PE were analyzed using one-way ANOVA. The evolution of SBP, assessed by tail-cuff plethysmography, was analyzed using repeated measures ANOVA, followed by Tukey's post-hoc test. P < 0.05 was considered statistically significant. 3 Results Figure 1 shows an evaluation of SBP using TCP over 5 weeks of treatment with RO and/or physical exercise. A reduction in SBP was observed over the 5 weeks of treatment in the RO SHR and RO TSHR groups compared to the baseline measurement and to the SHR control group. In addition to the evaluation of SBP, heart rate (HR) was extracted from the plethysmography data. It was observed that, throughout the interventions, no differences were found between or within groups. Figure 2 A illustrates the representative tracing of each experimental group from the PAP. A 60-second segment was taken from the baseline period to demonstrate the average pressure levels. Similar to the representative tracing, other variables were extracted from the PAP, such as MAP, and HR (Fig. 2 B). In the initial analysis, it was observed that MAP was higher in the SHR control group compared to the Wistar control group. However, the RO SHR and RO TSHR groups showed reduced levels of MAP, reinforcing the results seen in the TCP. Additionally, PE alone did not alter any of the baseline parameters of SHR. There was no difference in the baseline HR among the investigated groups. Figure 3 A presents the analyses of SBP variability in the TD, wherein the parameters SBP, SD, and variance were higher in the SHR control group compared to the Wistar group. Only the RO treatment attenuated the SBP analyzed in the TD when compared to the SHR control group. In the FD analysis, it was found that the LF of SBP was higher in the SHR control group compared to the Wistar group (Fig. 3 B). Neither the RO treatment nor PE altered the LF of SBP. Figure 4 A represents the results obtained from the HRV analysis in both the TD and FD. In the TD, there was no difference in pulse interval (ms) between the groups. The values for SD, variance, and RMSSD were lower in the SHR control group compared to the Wistar group, but the SD did not attain statistical significance. Only the treatment combining RO + PE was able to prevent the decrease in SD, variance, and RMSSD values. In the FD, there were no differences between the groups in absolute LF (ms²), normalized LF and HF (nu), and the LF/HF ratio. The absolute HF (ms²) was lower in the SHR control group compared to the Wistar group. Only the RO + PE treatment was able to prevent the decrease in absolute HF of SHR (Fig. 4 B). Figure 5 illustrates the data from the symbolic analysis, which investigates autonomic modulation through the percentiles of 0V, 1V, and 2V extracted from the pulse interval (PI). Sympathetic modulation, expressed by the 0V values, increased in the SHR control group compared to the Wistar group. Additionally, the 2V variable (which corresponds to vagal modulation) was decreased in the SHR control animals. Only the combination of RO + PE was able to prevent the increase in 0V and the reduction in 2V in the SHR animals. The 1V, representing sympathovagal modulation, showed no differences between the groups. Figure 6 A shows the baroreflex index response to the administration of vasoconstrictor phenylephrine (which induces bradycardic response) and vasodilator sodium nitroprusside (which induces tachycardic response), respectively. The results indicate that the bradycardic response was attenuated in the SHR animals compared to the Wistar group, and the different treatments did not reverse this condition. The tachycardic response was similar among the studied groups. Figure 6 B presents the evaluation of cardiac parasympathetic tone following the administration of methyl-atropine. The SHR control group exhibited lower parasympathetic tone compared to the Wistar group. Additionally, only the combined treatment of RO + PE was able to prevent the decline in parasympathetic tone. Finally, Fig. 6 C refers to the evaluation of the intrinsic heart rate (IHR). In this analysis, it was observed that IHR was lower in the SHR control group compared to the Wistar group. The treatment with RO and/or PE did not alter this response in SHR. Discussion This study is the first to evaluate the chronic effects of the monoterpene RO in combination with PE on cardiovascular alterations, particularly focusing on autonomic and hemodynamic aspects. RO has been reported to be a potential antihypertensive tool, as shown in the study by Santos et al. [ 9 ], which evaluated the acute hypotensive effect of RO at doses of 1.25, 2.5, and 5.0 mg/kg (i.v.). In the present study, it was observed that after the first week of treatment (oral administration), there was a decrease in SBP, as measured by TCP, in the RO SHR and RO TSHR groups. This decrease was sustained until the last SBP measurement, a result that partially aligns with observations from our previous studies (unpublished data). In those observations, a significant decrease in SBP was noted in SHR up to the 7th day, but returned to pretreatment values on the 14th day [sub chronic protocol of 14 days, oral administration, 50 mg/kg diluted with Tween-80 (polyoxyethylene sorbitan monolaurate, 5% + saline 0.9%)]. Based on this preliminary result, we opted in the present study to increase the dose of RO (100 mg/kg) and dilute the compound in Tween-20 (5%), considering how well tween-20 stabilizes hydrophobic compounds [ 27 ]. Consequently, we recorded a decrease in SBP that lasted for the entire 34 days of treatment. Furthermore, it is essential to emphasize that TCP is an indirect recording method that includes factors that can be stressful to the animal, potentially compromising the accuracy of the test. Therefore, it is challenging to predict the extent to which these factors influenced the results obtained when monitoring the evolution of SBP and HR throughout the treatment [ 28 ]. Figure 2 A displays representative recordings extracted from PAP over 1 minute, to compare PAP levels across different groups. The results of MAP in the SHR control (Fig. 2 B) reflect the well-established arterial hypertension [ 29 , 30 ] compared to Wistar rats [ 31 ]. In addition, MAP was significantly reduced in both RO alone and RO + PE groups. On the other hand, PE alone did not attenuate the parameters of BP. These results suggest that RO is a promising antihypertensive agent, as groups treated with RO showed a reduction in BP assessed through TCP and PAP. Thus, the findings reinforce the effectiveness of RO treatment on BP, and express the inefficiency of the chosen physical exercise protocol for this study, since no reduction in blood pressure levels was observed in the TSHR group, similar to the HR variable, in which no difference was observed between the studied groups. Therefore, the ineffectiveness of the swimming protocol is attributed to intrinsic training variables such as intensity and volume, which did not change after the 1-week training period. Consequently, it is hypothesized that better adjustment of training variables would lead to better responses in MAP, and HC, which means a dose-response relationship [ 18 , 32 , 33 , 34 ]. The variability of heart rate and SBP has been reported as an important predictor metric capable of assessing autonomic control over the heart and blood vessels, as well as evaluating the imminence of various pathophysiological conditions such as cardiovascular diseases, diabetic cardiovascular autonomic neuropathy, renal dysfunction, among others. Therefore, this variability is recognized as a crucial health indicator [ 26 , 35 , 36 ]. In this context, when analyzing SBP in the TD and FD (Fig. 3 A and B, respectively), differences were observed between the control groups in all analyzed parameters. Furthermore, consistent with the findings of this study, the RO SHR group showed a significant reduction in SBP in the time domain as well as a clear improvement trend in the LF component of BP on the frequency domain. This result suggests that the reduction in BP observed in SHR may be partly due to the attenuation of sympathetic activity to the vascular bed. A similar result has been demonstrated by our laboratory with intravenous administration of RO [ 9 ], in which we observed a decrease in MAP and LF of SBP induced by RO at doses of 1.25, 2.5, and 5 mg/kg (i.v.). However, after 15 minutes, the decrease in MAP at the two lower doses was independent of a significant attenuation of LF of SBP [ 9 ]. Thus, the results of the two lower doses align with the findings of the current study, in which we observed a decrease in MAP independent of the significant attenuation of LF of SBP. Pulse interval (PI), as described by Mejía-Mejía et al. [ 37 ], is an important measure in assessing HRV in both TD and FD. In SHR, PI (Fig. 4 A, right graph) did not show significant differences between the analyzed groups, but SD, variance, and RMSSD showed a clear trend of decrease compared to the Wistar group, still, SD did not attain significance. These aforementioned parameters reflect parasympathetic cardiac modulation, which indicates an already expected vagal attenuation in SHR [ 29 , 38 ]. However, only the RO + PE treatment was able to prevent the decline in parasympathetic modulation, as an increase in SD, variance, and RMSSD was noted in the RO TSHR group. Thus, it can be inferred that despite the limitations regarding the training protocol, RO treatment combined with PE was able to promote improvement in SNP, considering that RMSSD reflects cardiac vagal activity [ 35 ]. Still regarding our study limitations, we recognize that while this study provides valuable insights into the potential benefits of combining RO with physical exercise for improving cardiovascular and autonomic functions in hypertensive rats, several other limitations should be acknowledged. First, the study was conducted on animal models, specifically SHR, which, while informative, may not fully replicate the complexities of human hypertension. Further research involving clinical trials is necessary to confirm the translatability of these findings to human subjects. Second, the physical exercise protocol used in this study was limited to swimming, which may not represent other forms of exercise that could have different impacts on cardiovascular and autonomic functions. Exploring various exercise modalities and intensities could provide a more comprehensive understanding of the synergistic effects of RO and exercise. Third, the study duration was relatively short, lasting only five weeks. Longer-term studies are needed to evaluate the sustained effects and potential long-term benefits or drawbacks of this combined treatment approach. Additionally, while the study focused on hemodynamic and autonomic parameters, other relevant physiological and molecular mechanisms underlying the observed effects were not explored in detail. Future research should include a broader range of biomarkers and mechanistic studies to elucidate the pathways involved. Finally, the exact dosage and formulation of RO were based on previous animal studies, and optimal dosing for humans remains to be determined. These limitations highlight the need for further research to validate and expand upon the findings presented in this study. As described previously, the LF component of HRV is associated with sympathetic autonomic modulation, while the HF component is related to vagal modulation [ 36 , 39 , 40 ]. In evaluating LF in the FD, both LF (ms²) and LF (nu) showed no differences between the experimental groups; however, there was a noticeable trend towards a reduction in LF (nu) in the RO TSHR group. Additionally, when analyzing HF (ms²), an improvement in vagal modulation was observed in the RO TSHR group, consistent with the findings related to parasympathetic tone, SD, variance, and RMSSD in this study. Ultimately, no differences were observed in the LF/HF ratio. Autonomic modulation was also assessed using symbolic analysis, in which an increase at 0V and a decrease at 2V were observed in SHR, indicating sympathetic dominance and reduced parasympathetic modulation, respectively. These findings align with other studies showing autonomic imbalance in SHR [ 38 ]. Similarly, to the findings in the FD, only the RO + PE group was able to promote an improvement in the autonomic nervous system by reducing 0V and increasing 2V. Therefore, the results of the analysis of autonomic variability via linear and non-linear methods, as well as parasympathetic tone, illustrate that the combination of RO with PE was the only treatment capable of inducing hypotension accompanied by an improvement in the cardiac autonomic nervous system. This indicates the need to explore other exercise protocols in combination with RO to further investigate the potential association of this dual intervention in controlling systemic arterial hypertension. Results also showed attenuated baroreflex in SHR (after phenylephrine administration) compared to Wistar, as expected. However, none of the treatments were effective in improving baroreflex sensitivity. Furthermore, other studies have reported improvements in this parameter, since it was possible to observe enhancement of baroreflex in animals undergoing aerobic exercise in combination with antihypertensive drugs [ 28 , 41 ]. Given these findings, we hypothesize that the exercise intervention chosen for this study was not effective in improving this variable. Analyzing the IHR and other studies that have addressed this parameter, it is worth mentioning that the decrease in IHR in SHR animals is consistent with sympathetic hyperactivity, along with electrical and structural remodeling of individual cells in the cardiac atrial pacemaker [ 42 , 43 ]. Moreover, the interventions were not effective in improving IHR. The main findings of the current study are illustrated in Fig. 7 . Conclusion In summary, this was the first study to evaluate the hypotensive activity of chronic treatment with the monoterpene Rose Oxide in combination with physical exercise, showing that RO was able to reduce blood pressure levels in SHR both in TCP and PAP, along with improvements observed in the autonomic control of blood pressure through analysis of HRV and autonomic tone. RO is, therefore, a potential antihypertensive tool. Adjustments of the physical training variables and dosage could lead to promising results. ‌ Declarations Funding This study was supported by grants CNPq 409109/2018-5, 306987/2021-0, 423999/2021-4 and FAPEPI 00110.000235/2022-78. Author Contribution Juliana A. da Silva, Samuel de S. P. Araújo, Ana Flávia M. da Silva, José Guilherme V. de Assunção, Pâmela de S. Santos, José L. P. Júnior, Carlos Eduardo S. Reis, Liana de M. Santana, Regina G. Silva, Ariell A. de Oliveira, Francisca V. de Sousa Nunes: Performed the experimental and treatment protocol. Juliana A. da Silva, Samuel de S. P. Araújo and João Paulo J. Sabino wrote the main manuscript.Aldeidia P. de Oliveira, Damião P. de Sousa, Renato N. Soriano, Luiz G. S. Branco, Helio C. Salgado, João Paulo J. Sabino Analysis of resultsAll authors reviewed the manuscript Acknowledgments This study was supported by grants CNPq 409109/2018-5, 306987/2021-0, 423999/2021-4 and FAPEPI 00110.000235/2022-78. References AL GHORANI, H. et al. 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Respiratory Physiology & Neurobiology, v. 231, p. 21–27, 1 set. 2016. https://doi.org/10.1016/j.resp.2019.03.001 . REZENDE, L. M. T. DE et al. Is the Wistar Rat a more Suitable Normotensive Control for SHR to Test Blood Pressure and Cardiac Structure and Function? International Journal of Cardiovascular Sciences, 26 jul. 2021. https://doi.org/10.36660/ijcs.20200367 NEVES, R. V. P. et al. Treinamento de força em ratos espontaneamente hipertensos com hipertensão arterial grave. Arquivos Brasileiros de Cardiologia, v. 106, n. 3, p. 201–209, 2016. https://doi.org/10.5935/abc.20160019 . ‌HEGDE, S. M.; SOLOMON, S. D. Influence of Physical Activity on Hypertension and Cardiac Structure and Function. Current Hypertension Reports, v. 17, n. 10, 2015 https://doi.org/10.1007/s11906-015-0588-3 DIAZ, K. M.; SHIMBO, D. Physical Activity and the Prevention of Hypertension. Current Hypertension Reports, v. 15, n. 6, p. 659–668, 20 set. 2013. https://doi.org/10.1007/s11906-013-0386-8 . YUGAR, L. B. T. et al. The Role of Heart Rate Variability (HRV) in Different Hypertensive Syndromes. Diagnostics (Basel, Switzerland), 13(4), 785,2023. https://doi.org/10.3390/diagnostics13040785 . TIWARI, R. et al. Analysis of Heart Rate Variability and Implication of Different Factors on Heart Rate Variability. Current Cardiology Reviews, v. 17, n. 5, 31 dez. 2020. https://doi.org/10.2174/1573403X16999201231203854 . ‌ MEJÍA-MEJÍA, E. et al. Pulse rate variability in cardiovascular health: a review on its applications and relationship with heart rate variability. Physiol Meas. 2020;41(7):07TR01. https://doi.org/10.1088/1361-6579/ab998c VIEIRA-ROCHA, M. S. et al. Elevated Vascular Sympathetic Neurotransmission and Remodelling Is a Common Feature in a Rat Model of Foetal Programming of Hypertension and SHR. Biomedicines, v. 10, n. 8, p. 1902, 1 ago. 2022. https://doi.org/10.3390/biomedicines10081902 . SILVA, L. E. V. et al. Comparison between spectral analysis and symbolic dynamics for heart rate variability analysis in the rat. Scientific Reports, v. 7, n. 1, 16 ago. 2017. https://doi.org/10.1038/s41598-017-08888-w . VANDERLEI, L. C. M. et al. Noções básicas de variabilidade da frequência cardíaca e sua aplicabilidade clínica. Revista Brasileira de Cirurgia Cardiovascular, v. 24, n. 2, p. 205–217, jun. 2009. https://doi.org/10.1590/s0102-76382009000200018 . ‌FERREIRA-JUNIOR, N. C. et al. Exercise training increases GAD65 expression, restores the depressed GABA A receptor function within the PVN and reduces sympathetic modulation in hypertension. Physiological reports, v. 7, n. 13, 1 jul. 2019. https://doi.org/10.14814/phy2.14107 . SAYIN, H. et al. Assessment of cardiac autonomic tone in conscious rats. Autonomic Neuroscience, Elsevier B.V., v. 194, p. 26–31, 1 2016. ISSN 15660702. https://doi.org/10.1016/j.autneu.2015.12.007 . ‌RODRIGUES, J. Q. D. et al. Intrinsic adaptation of SHR right atrium reduces heart rate. Journal of Cardiovascular Pharmacology, v. 74, n. 6, p. 542–548, 2019. https://doi.org/10.1097/FJC.0000000000000746 . Additional Declarations No competing interests reported. Cite Share Download PDF Status: Published Journal Publication published 30 Oct, 2024 Read the published version in Pflügers Archiv - European Journal of Physiology → Version 1 posted Editorial decision: Revision requested 20 Sep, 2024 Reviews received at journal 20 Sep, 2024 Reviewers agreed at journal 11 Sep, 2024 Reviewers agreed at journal 10 Sep, 2024 Reviews received at journal 02 Sep, 2024 Reviewers agreed at journal 27 Aug, 2024 Reviewers invited by journal 26 Aug, 2024 Editor assigned by journal 21 Aug, 2024 Submission checks completed at journal 21 Aug, 2024 First submitted to journal 19 Aug, 2024 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4939277","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":356415310,"identity":"cb85cd9d-4e33-4b7b-9a49-98eb05bd166f","order_by":0,"name":"Juliana A. da Silva","email":"","orcid":"","institution":"Federal University of Piauí","correspondingAuthor":false,"prefix":"","firstName":"Juliana","middleName":"A. da","lastName":"Silva","suffix":""},{"id":356415312,"identity":"dacfe5e0-0d8b-49ed-bd56-29b632870f1c","order_by":1,"name":"Samuel S. P. Araújo","email":"","orcid":"","institution":"Federal University of Piauí","correspondingAuthor":false,"prefix":"","firstName":"Samuel","middleName":"S. P.","lastName":"Araújo","suffix":""},{"id":356415315,"identity":"d09dfd58-8d6e-4188-a778-f6435a4357b2","order_by":2,"name":"Ana Flávia M. da Silva","email":"","orcid":"","institution":"Federal University of Piauí","correspondingAuthor":false,"prefix":"","firstName":"Ana","middleName":"Flávia M. da","lastName":"Silva","suffix":""},{"id":356415316,"identity":"dc4190d4-347f-489a-8f6c-d2b8263505a0","order_by":3,"name":"José Guilherme V. de Assunção","email":"","orcid":"","institution":"Federal University of Piauí","correspondingAuthor":false,"prefix":"","firstName":"José","middleName":"Guilherme V.","lastName":"de Assunção","suffix":""},{"id":356415318,"identity":"9219702b-42ba-40ce-8f0d-48290c87989c","order_by":4,"name":"Pâmela S. Santos","email":"","orcid":"","institution":"Federal University of Piauí","correspondingAuthor":false,"prefix":"","firstName":"Pâmela","middleName":"S.","lastName":"Santos","suffix":""},{"id":356415319,"identity":"43d1d2cd-73e5-41a5-97e4-d0e8cbe480a6","order_by":5,"name":"José L. P. Júnior","email":"","orcid":"","institution":"Federal University of Piauí","correspondingAuthor":false,"prefix":"","firstName":"José","middleName":"L. P.","lastName":"Júnior","suffix":""},{"id":356415322,"identity":"206652f7-03f5-47c9-8ec0-2430441e4eb4","order_by":6,"name":"Carlos Eduardo S. Reis","email":"","orcid":"","institution":"Federal University of Piauí","correspondingAuthor":false,"prefix":"","firstName":"Carlos","middleName":"Eduardo S.","lastName":"Reis","suffix":""},{"id":356415324,"identity":"16641a40-a449-4dc2-b1ae-7ff0219adbab","order_by":7,"name":"Liana M. 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Sousa Nunes","email":"","orcid":"","institution":"Federal University of Piauí","correspondingAuthor":false,"prefix":"","firstName":"Francisca","middleName":"V. Sousa","lastName":"Nunes","suffix":""},{"id":356415330,"identity":"89062052-5683-4793-81ed-b024f5112c40","order_by":11,"name":"Aldeidia P. Oliveira","email":"","orcid":"","institution":"Federal University of Piauí","correspondingAuthor":false,"prefix":"","firstName":"Aldeidia","middleName":"P.","lastName":"Oliveira","suffix":""},{"id":356415331,"identity":"7f37062c-2557-4cf2-847b-5f675e24d5d4","order_by":12,"name":"Damião P. Sousa","email":"","orcid":"","institution":"Federal University of Paraíba","correspondingAuthor":false,"prefix":"","firstName":"Damião","middleName":"P.","lastName":"Sousa","suffix":""},{"id":356415332,"identity":"beb85c06-f452-44e2-8547-13ec0e1ec228","order_by":13,"name":"Renato N. Soriano","email":"","orcid":"","institution":"Federal University of Juiz de Fora, Brazil","correspondingAuthor":false,"prefix":"","firstName":"Renato","middleName":"N.","lastName":"Soriano","suffix":""},{"id":356415333,"identity":"8ccca1d2-a474-4015-8daa-2313cfd25efe","order_by":14,"name":"Luiz G. S. Branco","email":"","orcid":"","institution":"University of São Paulo","correspondingAuthor":false,"prefix":"","firstName":"Luiz","middleName":"G. S.","lastName":"Branco","suffix":""},{"id":356415334,"identity":"8ebdd179-f0de-4402-a563-0d0babb5f361","order_by":15,"name":"Helio C. Salgado","email":"","orcid":"","institution":"University of São Paulo","correspondingAuthor":false,"prefix":"","firstName":"Helio","middleName":"C.","lastName":"Salgado","suffix":""},{"id":356415335,"identity":"0af97e48-99d5-4ed8-9320-868eaf3b68a4","order_by":16,"name":"João Paulo Jacob Sabino","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA+0lEQVRIiWNgGAWjYBACxgYGBmYgncAGZD9gsGHgAXIMwMLEaGE2YEgjQgsIgLUAMZsEUAsDQS3M7YcfPi74Y5fHJ918rJon4Y4MA3vzNgnGHfdwO6wnzdh4ZltyMZvMsbTbPAnPeBh4jpVJMJ4pxuOXHDZp3gbmxDaJHLPbvD8O8zAAGRKMbQm4tfS/YZPm+VMP1lLMkwDUIv+GgJYZQFt42A6DtTCDtUjwENLyDOSX48VsEmnJknOAfmHjSSu2SDyDW4thfzIoxKrz5GckH/zwJuGOPT/74Y03Pu7Ao6UBlX+AgQ1E4dbAwCCPxj+AR+0oGAWjYBSMVAAAnmFNc65SzrsAAAAASUVORK5CYII=","orcid":"","institution":"Federal University of Piauí","correspondingAuthor":true,"prefix":"","firstName":"João","middleName":"Paulo Jacob","lastName":"Sabino","suffix":""}],"badges":[],"createdAt":"2024-08-19 14:21:19","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4939277/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4939277/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s00424-024-03035-7","type":"published","date":"2024-10-30T15:56:53+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":66668185,"identity":"6e852b90-5249-4fa9-9cc5-67c750d14204","added_by":"auto","created_at":"2024-10-15 09:54:23","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":18870,"visible":true,"origin":"","legend":"\u003cp\u003eSystolic Blood Pressure Variation (ΔSBP) and Heart Rate Variation (ΔHR) of rats obtained by tail plethysmography at weeks 0, 1, 3, and 5. Groups Wistar (n=10), SHR (n=14), RO SHR (n=14), TSHR (n=9), and RO TSHR (n=9) were compared using two-way repeated measures ANOVA. * p \u0026lt; 0.05 compared to day 0 within the same group; # p \u0026lt; 0.05 compared to the SHR vehicle groupa\u003c/p\u003e","description":"","filename":"FIG1.png","url":"https://assets-eu.researchsquare.com/files/rs-4939277/v1/143d8a8a96e077c2544b77ad.png"},{"id":66667159,"identity":"ba4c00d4-4e66-4dd3-a82b-f15cb34a02f1","added_by":"auto","created_at":"2024-10-15 09:46:23","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":41428,"visible":true,"origin":"","legend":"\u003cp\u003e(a) Representative records of pulsatile arterial pressure (PAP), and mean arterial pressure (PAM; white traces) of the experimental groups under baseline conditions. (b) Hemodynamic parameters obtained by PAP: Systolic Arterial Pressure (PAS), Diastolic Arterial Pressure (PAD), Mean Arterial Pressure (PAM), and Heart Rate (HR) of Wistar rats (n=8), SHR (n=7), RO SHR (n=11), TSHR (n=8), RO TSHR (n=8) groups. * p \u0026lt; 0.05 compared to the Wistar group; # p \u0026lt; 0.05 compared to the SHR group\u003c/p\u003e","description":"","filename":"FIG2.png","url":"https://assets-eu.researchsquare.com/files/rs-4939277/v1/e0ccc7a980b5555bedc3ecb9.png"},{"id":66667160,"identity":"f02b1d6d-79ea-4811-9151-d63f1ed9e945","added_by":"auto","created_at":"2024-10-15 09:46:24","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":31714,"visible":true,"origin":"","legend":"\u003cp\u003eVariability of Systolic Arterial Pressure (SAP) assessed in a) Time domains; b) Frequency domains: absolute values (abs) of low-frequency power LF. * p \u0026lt; 0.05 compared to the Wistar group\u003c/p\u003e","description":"","filename":"FIG3.png","url":"https://assets-eu.researchsquare.com/files/rs-4939277/v1/656f0b759aef42c748dc4c72.png"},{"id":66668186,"identity":"a7460c10-0f28-40a4-bd3f-f712607118f6","added_by":"auto","created_at":"2024-10-15 09:54:24","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":64273,"visible":true,"origin":"","legend":"\u003cp\u003eHeart Rate Variability (HRV) in a) Time domain: standard deviation (SD), variance, and the square root of the mean of the sum of the squares of differences between adjacent normal RR intervals (RMSSD); b) Frequency domain: absolute values (abs) of low-frequency power (LF) and high-frequency power (HF), normalized power (nu) of spectra in each frequency band, and the LF/HF ratio of Systolic Arterial Pressure (SAP) in the groups. * p \u0026lt; 0.05 compared to the Wistar group; # p \u0026lt; 0.05 compared to the SHR control group\u003c/p\u003e","description":"","filename":"FIG4.png","url":"https://assets-eu.researchsquare.com/files/rs-4939277/v1/36e6d38c5434725ea6afce8b.png"},{"id":66667162,"identity":"a6906df9-8329-48ec-a08a-387796fda57b","added_by":"auto","created_at":"2024-10-15 09:46:24","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":32768,"visible":true,"origin":"","legend":"\u003cp\u003eSymbolic analysis of percentile of sympathetic modulation 0V% and 1V% and parasympathetic 2V%. * p \u0026lt; 0.05 compared to the Wistar group; # p \u0026lt; 0.05 compared to the SHR group; π p \u0026lt; 0.05 compared to the TSHR group\u003c/p\u003e","description":"","filename":"FIG5.png","url":"https://assets-eu.researchsquare.com/files/rs-4939277/v1/52495b932ffe163e2911885f.png"},{"id":66667163,"identity":"d58f5f06-c7f2-4ede-bbaf-e72043539254","added_by":"auto","created_at":"2024-10-15 09:46:24","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":44298,"visible":true,"origin":"","legend":"\u003cp\u003e(a) Baroreflex index in response to phenylephrine and baroreflex response to NPS; (b) Parasympathetic cardiac tone after administration of atropine; (c) Intrinsic heart rate (FIMP) after administration of atropine and propranolol, obtained by monitoring PAP. The groups Wistar (n=8), SHR (n=7), RO SHR (n=11), TSHR (n=8), and RO TSHR (n=8) were compared by one-way ANOVA. * p \u0026lt; 0.05 compared to the Wistar group; # p \u0026lt; 0.05 compared to the SHR group\u003c/p\u003e","description":"","filename":"FIG6.png","url":"https://assets-eu.researchsquare.com/files/rs-4939277/v1/007614bf839b732d986cfc81.png"},{"id":66667164,"identity":"3444f430-beb1-47e2-8feb-6a58440f7166","added_by":"auto","created_at":"2024-10-15 09:46:24","extension":"jpeg","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":1821335,"visible":true,"origin":"","legend":"\u003cp\u003eSchematic drawing of the findings showing the effect of chronic rose oxide and exercise on systolic arterial pressure, mean arterial pressure and heart rate variability in Wistar and SHR rats. Created with \u003ca href=\"https://biorender.com/\"\u003eBioRender.com\u003c/a\u003e\u003c/p\u003e","description":"","filename":"Figure7.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-4939277/v1/9058039173f11f83f33e3b24.jpeg"},{"id":68206343,"identity":"6dca8a87-fc82-45e2-a0b2-74c832b14195","added_by":"auto","created_at":"2024-11-04 16:31:26","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2558563,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4939277/v1/402da68e-a876-4f12-8665-206899ceddd5.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Chronic Rose Oxide and Exercise Synergistically Modulate Cardiovascular and Autonomic Functions in Hypertensive Rats","fulltext":[{"header":"1 Introduction","content":"\u003cp\u003eAnalyzing the global context in recent years, there has been a noticeable increase in the number of people affected by some type of cardiovascular disease (CVD). In this alarming scenario, arterial hypertension (AH) is among the most prevalent CVD [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. It is estimated that approximately one-third of the world population has blood pressure levels\u0026thinsp;\u0026ge;\u0026thinsp;140/90 mmHg, characterizing AH, whose treatment costs are high and mortality rate has risen by 56.1% in recent years [\u003cspan additionalcitationids=\"CR3 CR4\" citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eFurthermore, unbalances in cardiac autonomic function, such as arterial baroreflex dysfunction and severe sympathetic hyperactivity combined with a decrease in parasympathetic activity, which characterizes a sympathetic-vagal imbalance, are known to exist in the pathogenesis of AH [\u003cspan additionalcitationids=\"CR7\" citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. In this context, the evaluation of markers of cardiac autonomic control, such as baroreflex and parasympathetic tone, is of utmost importance for the monitoring and control of this disorder. In this respect, Heart Rate Variability (HRV) is a widely used mathematical model to infer autonomic activity and it is used in studies with both humans and animals [\u003cspan additionalcitationids=\"CR10\" citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eInterestingly, the secondary metabolites called terpenes are promising therapeutic molecules in controlling AH [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. A subdivision of terpenes called monoterpenes has gained prominence due to their vast diversity, abundance, and pharmacological properties in the circulatory system [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e] In this perspective, Rose Oxide (RO), a tetrahydropyran of the monoterpene class, has shown therapeutic potential in the treatment of AH, considering its anti-inflammatory and antihypertensive actions [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e, \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e] .\u003c/p\u003e \u003cp\u003eOn the other hand, besides pharmacological interventions, both resistance exercise and aerobic physical exercise (PE) are characterized as an excellent alternative approach capable of delaying AH development and enhancing its treatment, which potentially reduces mortality rates in some clinical outcomes. Moreover, it is known that PE promotes numerous physiological adaptations, such as improved endothelial function, glucose uptake, parasympathetic autonomic activity, and angiogenesis, neutralizes the generation of reactive oxygen species and promotes cardiac hypertrophy, among other benefits [\u003cspan additionalcitationids=\"CR17\" citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThus, it is believed that combining pharmacological intervention with PE in the treatment of AH could be an effective therapeutic strategy for controlling this disease, thereby promoting better life expectancy, and reducing the amount or dosage of medications, directly influencing efficacy [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]. Therefore, our objective was to evaluate the chronic hypotensive activity of RO in combination with PE (swimming exercise). For this purpose, we used an inbred genetic model of experimental hypertension (Spontaneously Hypertensive Rats, SHR) to mimic AH that is observed in humans [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e, \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]\u003c/p\u003e"},{"header":"2 Methodology","content":"\u003cp\u003e2.1. Animals\u003c/p\u003e \u003cp\u003eExperiments were conducted with Wistar rats and SHR provided by the Federal University of Piau\u0026iacute; (UFPI). For the criteria of animal use and environmental control, we followed the same protocol proposed by Santos et al. [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. It is worth noting that all experimental procedures adhered to the guidelines of the National Institutes of Health for the care and use of laboratory animals and were approved by the Ethics Committee on Animal Use of the Federal University of Piau\u0026iacute; (CEUA/UFPI # 563/19).\u003c/p\u003e \u003cp\u003e2.2. Experimental Groups\u003c/p\u003e \u003cp\u003eThe animals were divided into five groups, as follows: 1) Wistar \u0026ndash; control; 2) SHR \u0026ndash; control; 3) RO SHR \u0026ndash; administration of RO; 4) TSHR \u0026ndash; aerobic training intervention; 5) RO TSHR \u0026ndash; a combination of aerobic training and RO administration.\u003c/p\u003e \u003cp\u003e2.3. Reagents\u003c/p\u003e \u003cp\u003e(+)-rose oxide (tetrahydro-4-methyl-2-(2-methylprop-1-enyl) pyran) was acquired from Sigma-Aldrich (Barueri, SP, Brazil; 99.0% purity), diluted in 2% Tween 20 P.S. (Vetec Qu\u0026iacute;mica Fina Ltda.) and saline (0.9% sodium chloride). The dose used to evaluate the effect of RO was 100 mg/kg (adapted from Maia et al., 2021 [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]; Nonato et al., 2012 [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]).\u003c/p\u003e \u003cp\u003eDuring the pulsatile arterial pressure (PAP) monitoring, the following drugs were used: phenylephrine (4 \u0026micro;g/kg i.v.), sodium nitroprusside [NPS (8 \u0026micro;g/kg i.v.)], methyl-atropine (2 mg/kg, i.v.), and propranolol (4 mg/kg, i.v.), all acquired from Sigma-Aldrich, St. Louis, MO, USA. For animal euthanasia, sodium thiopental (Crist\u0026aacute;lia Prod. Qu\u0026iacute;m. Farm. Ltda; injectable solution; 100 mg/kg; i.v.) was used at the end of the PAP recording, as established by Resolution No. 1,000 of May 11, 2012, of the Brazilian Federal Council of Veterinary Medicine \u0026ndash; CFMV.\u003c/p\u003e \u003cp\u003e2.4. Tail-cuff plethysmography (TCP)\u003c/p\u003e \u003cp\u003eTo monitor the evolution of SBP in the animals during treatment, blood pressure (BP) monitoring was conducted using tail-cuff plethysmography (ADInstruments, Australia), following the protocol described by Silva et al. [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. Plethysmography was performed at 4 time points: day 0 (baseline), week 1, week 3, and week 5 of treatment with RO or vehicle.\u003c/p\u003e \u003cp\u003e2.5. Gavage\u003c/p\u003e \u003cp\u003eRO or vehicle was administered orally using a gavage needle for 34 consecutive days, according to the protocol by Machholz et al. [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e2.6. Swimming training\u003c/p\u003e \u003cp\u003eAerobic swimming training protocol, adapted from Rocha [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e], was employed. All training groups initially underwent an adaptation period, which involved swimming in a cylindrical tank (80.0 cm in diameter, 120 cm deep) filled with thermo-neutral water (temperature of 30\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C) at a depth of approximately 100 cm. To minimize animal stress, the free-swimming time interval gradually increased as follows: 1st day \u0026minus;\u0026thinsp;15 min; 2nd day \u0026minus;\u0026thinsp;30 min; 3rd day \u0026minus;\u0026thinsp;45 min; and 4th day \u0026minus;\u0026thinsp;60 min.\u003c/p\u003e \u003cp\u003eSubsequently, the rats swam for 4 weeks, swimming for 60 minutes per day, 5 times a week.\u003c/p\u003e \u003cp\u003e2.7. Surgical procedure for femoral artery and vein cannulation\u003c/p\u003e \u003cp\u003eDuring the surgical procedure, the anesthetics ketamine (33.33 mg/kg) and xylazine (13.3 mg/kg) were used. The vessel cannulation procedure was performed as Santos et al. [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e] and Oliveira [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e] described.\u003c/p\u003e \u003cp\u003e2.8. Pulsatile arterial pressure monitoring\u003c/p\u003e \u003cp\u003eDuring the PAP recording, the arterial catheter was connected to a pressure transducer (MLT0380/D, ADInstruments, Sydney, Australia). The signal, after proper amplification (ML110 Amplifier, ADInstruments, Sydney, Australia), was digitized through an analog-to-digital interface (ML866, ADInstruments, Sydney, Australia) and displayed at 2000 Hz on a microcomputer. Additionally, from the PAP recording, it was possible to extract mean arterial pressure (MAP) and heart rate (HR). The venous cannula was connected to a PE tube extension to facilitate drug administration, aiming to minimize animal stress during monitoring.\u003c/p\u003e \u003cp\u003eMonitoring was divided into three parts: a period of animal acclimatization in the system (60 minutes), followed by baseline blood pressure recording (30 minutes), and concluding with the administration of vasoactive compounds.\u003c/p\u003e \u003cp\u003e2.9. Heart rate and systolic arterial pressure (SAP) variability\u003c/p\u003e \u003cp\u003eAnalysis of pulse interval (PI) and SAP variability was conducted considering both time-domain (TD) and frequency-domain (FD) parameters using CardioSeries software (v2.7- \u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eCardioSeries Software - CardioSeries Software (danielpenteado.com)\u003c/span\u003e). To collect data, segments were extracted from the baseline period of pulsatile arterial pressure (PAP) monitoring.\u003c/p\u003e \u003cp\u003eIn the time domain, SAP variability was quantified using mean and standard deviation (SD), while PI was analyzed using mean and the square root of the mean of the sum of the squares of the differences between successive PI values (RMSSD). Regarding spectral analysis in the frequency domain, time series of SAP and PI were interpolated at 10 Hz and divided into continuous segments of 512 beats with 50% overlap. Each segment underwent Hanning windowing, and spectral analysis was performed using the Fast Fourier Transform (FFT). Oscillatory components were quantified in low-frequency (LF: 0.2\u0026ndash;0.75 Hz) and high-frequency (HF: 0.75\u0026ndash;3 Hz) bands. To assess sympathovagal balance, the LF/HF ratio was also calculated. Furthermore, the relative (%) and absolute power of the spectra within each frequency band were analyzed (Santos et al. 2021 [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e] protocol; Sabino et al. 2013 [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]).\u003c/p\u003e \u003cp\u003e2.10. Statistical analysis\u003c/p\u003e \u003cp\u003eThe results are expressed as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SEM (Standard Error of the Mean). Statistical analyses were performed using SigmaPlot software (version 11.0). Baseline hemodynamic values and the alterations induced by RO and/or PE were analyzed using one-way ANOVA. The evolution of SBP, assessed by tail-cuff plethysmography, was analyzed using repeated measures ANOVA, followed by Tukey's post-hoc test. P\u0026thinsp;\u0026lt;\u0026thinsp;0.05 was considered statistically significant.\u003c/p\u003e"},{"header":"3 Results","content":"\u003cp\u003eFigure \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e shows an evaluation of SBP using TCP over 5 weeks of treatment with RO and/or physical exercise. A reduction in SBP was observed over the 5 weeks of treatment in the RO SHR and RO TSHR groups compared to the baseline measurement and to the SHR control group.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eIn addition to the evaluation of SBP, heart rate (HR) was extracted from the plethysmography data. It was observed that, throughout the interventions, no differences were found between or within groups.\u003c/p\u003e \u003cp\u003eFigure \u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eA illustrates the representative tracing of each experimental group from the PAP. A 60-second segment was taken from the baseline period to demonstrate the average pressure levels. Similar to the representative tracing, other variables were extracted from the PAP, such as MAP, and HR (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eB). In the initial analysis, it was observed that MAP was higher in the SHR control group compared to the Wistar control group. However, the RO SHR and RO TSHR groups showed reduced levels of MAP, reinforcing the results seen in the TCP. Additionally, PE alone did not alter any of the baseline parameters of SHR. There was no difference in the baseline HR among the investigated groups.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eFigure \u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eA presents the analyses of SBP variability in the TD, wherein the parameters SBP, SD, and variance were higher in the SHR control group compared to the Wistar group. Only the RO treatment attenuated the SBP analyzed in the TD when compared to the SHR control group. In the FD analysis, it was found that the LF of SBP was higher in the SHR control group compared to the Wistar group (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eB). Neither the RO treatment nor PE altered the LF of SBP.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eFigure \u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eA represents the results obtained from the HRV analysis in both the TD and FD. In the TD, there was no difference in pulse interval (ms) between the groups. The values for SD, variance, and RMSSD were lower in the SHR control group compared to the Wistar group, but the SD did not attain statistical significance. Only the treatment combining RO\u0026thinsp;+\u0026thinsp;PE was able to prevent the decrease in SD, variance, and RMSSD values. In the FD, there were no differences between the groups in absolute LF (ms\u0026sup2;), normalized LF and HF (nu), and the LF/HF ratio. The absolute HF (ms\u0026sup2;) was lower in the SHR control group compared to the Wistar group. Only the RO\u0026thinsp;+\u0026thinsp;PE treatment was able to prevent the decrease in absolute HF of SHR (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eB).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eFigure \u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e illustrates the data from the symbolic analysis, which investigates autonomic modulation through the percentiles of 0V, 1V, and 2V extracted from the pulse interval (PI). Sympathetic modulation, expressed by the 0V values, increased in the SHR control group compared to the Wistar group. Additionally, the 2V variable (which corresponds to vagal modulation) was decreased in the SHR control animals. Only the combination of RO\u0026thinsp;+\u0026thinsp;PE was able to prevent the increase in 0V and the reduction in 2V in the SHR animals. The 1V, representing sympathovagal modulation, showed no differences between the groups.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eFigure \u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eA shows the baroreflex index response to the administration of vasoconstrictor phenylephrine (which induces bradycardic response) and vasodilator sodium nitroprusside (which induces tachycardic response), respectively. The results indicate that the bradycardic response was attenuated in the SHR animals compared to the Wistar group, and the different treatments did not reverse this condition. The tachycardic response was similar among the studied groups.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eFigure \u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eB presents the evaluation of cardiac parasympathetic tone following the administration of methyl-atropine. The SHR control group exhibited lower parasympathetic tone compared to the Wistar group. Additionally, only the combined treatment of RO\u0026thinsp;+\u0026thinsp;PE was able to prevent the decline in parasympathetic tone.\u003c/p\u003e \u003cp\u003eFinally, Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eC refers to the evaluation of the intrinsic heart rate (IHR). In this analysis, it was observed that IHR was lower in the SHR control group compared to the Wistar group. The treatment with RO and/or PE did not alter this response in SHR.\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eThis study is the first to evaluate the chronic effects of the monoterpene RO in combination with PE on cardiovascular alterations, particularly focusing on autonomic and hemodynamic aspects. RO has been reported to be a potential antihypertensive tool, as shown in the study by Santos et al. [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e], which evaluated the acute hypotensive effect of RO at doses of 1.25, 2.5, and 5.0 mg/kg (i.v.).\u003c/p\u003e \u003cp\u003eIn the present study, it was observed that after the first week of treatment (oral administration), there was a decrease in SBP, as measured by TCP, in the RO SHR and RO TSHR groups. This decrease was sustained until the last SBP measurement, a result that partially aligns with observations from our previous studies (unpublished data). In those observations, a significant decrease in SBP was noted in SHR up to the 7th day, but returned to pretreatment values on the 14th day [sub chronic protocol of 14 days, oral administration, 50 mg/kg diluted with Tween-80 (polyoxyethylene sorbitan monolaurate, 5% + saline 0.9%)]. Based on this preliminary result, we opted in the present study to increase the dose of RO (100 mg/kg) and dilute the compound in Tween-20 (5%), considering how well tween-20 stabilizes hydrophobic compounds [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e]. Consequently, we recorded a decrease in SBP that lasted for the entire 34 days of treatment.\u003c/p\u003e \u003cp\u003eFurthermore, it is essential to emphasize that TCP is an indirect recording method that includes factors that can be stressful to the animal, potentially compromising the accuracy of the test. Therefore, it is challenging to predict the extent to which these factors influenced the results obtained when monitoring the evolution of SBP and HR throughout the treatment [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eFigure \u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eA displays representative recordings extracted from PAP over 1 minute, to compare PAP levels across different groups. The results of MAP in the SHR control (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eB) reflect the well-established arterial hypertension [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e, \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e] compared to Wistar rats [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]. In addition, MAP was significantly reduced in both RO alone and RO\u0026thinsp;+\u0026thinsp;PE groups. On the other hand, PE alone did not attenuate the parameters of BP. These results suggest that RO is a promising antihypertensive agent, as groups treated with RO showed a reduction in BP assessed through TCP and PAP. Thus, the findings reinforce the effectiveness of RO treatment on BP, and express the inefficiency of the chosen physical exercise protocol for this study, since no reduction in blood pressure levels was observed in the TSHR group, similar to the HR variable, in which no difference was observed between the studied groups. Therefore, the ineffectiveness of the swimming protocol is attributed to intrinsic training variables such as intensity and volume, which did not change after the 1-week training period. Consequently, it is hypothesized that better adjustment of training variables would lead to better responses in MAP, and HC, which means a dose-response relationship [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e, \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e, \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e, \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe variability of heart rate and SBP has been reported as an important predictor metric capable of assessing autonomic control over the heart and blood vessels, as well as evaluating the imminence of various pathophysiological conditions such as cardiovascular diseases, diabetic cardiovascular autonomic neuropathy, renal dysfunction, among others. Therefore, this variability is recognized as a crucial health indicator [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e, \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e, \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e]. In this context, when analyzing SBP in the TD and FD (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eA and B, respectively), differences were observed between the control groups in all analyzed parameters. Furthermore, consistent with the findings of this study, the RO SHR group showed a significant reduction in SBP in the time domain as well as a clear improvement trend in the LF component of BP on the frequency domain. This result suggests that the reduction in BP observed in SHR may be partly due to the attenuation of sympathetic activity to the vascular bed. A similar result has been demonstrated by our laboratory with intravenous administration of RO [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e], in which we observed a decrease in MAP and LF of SBP induced by RO at doses of 1.25, 2.5, and 5 mg/kg (i.v.). However, after 15 minutes, the decrease in MAP at the two lower doses was independent of a significant attenuation of LF of SBP [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. Thus, the results of the two lower doses align with the findings of the current study, in which we observed a decrease in MAP independent of the significant attenuation of LF of SBP.\u003c/p\u003e \u003cp\u003ePulse interval (PI), as described by Mej\u0026iacute;a-Mej\u0026iacute;a et al. [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e], is an important measure in assessing HRV in both TD and FD. In SHR, PI (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eA, right graph) did not show significant differences between the analyzed groups, but SD, variance, and RMSSD showed a clear trend of decrease compared to the Wistar group, still, SD did not attain significance. These aforementioned parameters reflect parasympathetic cardiac modulation, which indicates an already expected vagal attenuation in SHR [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e, \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e]. However, only the RO\u0026thinsp;+\u0026thinsp;PE treatment was able to prevent the decline in parasympathetic modulation, as an increase in SD, variance, and RMSSD was noted in the RO TSHR group. Thus, it can be inferred that despite the limitations regarding the training protocol, RO treatment combined with PE was able to promote improvement in SNP, considering that RMSSD reflects cardiac vagal activity [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eStill regarding our study limitations, we recognize that while this study provides valuable insights into the potential benefits of combining RO with physical exercise for improving cardiovascular and autonomic functions in hypertensive rats, several other limitations should be acknowledged. First, the study was conducted on animal models, specifically SHR, which, while informative, may not fully replicate the complexities of human hypertension. Further research involving clinical trials is necessary to confirm the translatability of these findings to human subjects. Second, the physical exercise protocol used in this study was limited to swimming, which may not represent other forms of exercise that could have different impacts on cardiovascular and autonomic functions. Exploring various exercise modalities and intensities could provide a more comprehensive understanding of the synergistic effects of RO and exercise. Third, the study duration was relatively short, lasting only five weeks. Longer-term studies are needed to evaluate the sustained effects and potential long-term benefits or drawbacks of this combined treatment approach. Additionally, while the study focused on hemodynamic and autonomic parameters, other relevant physiological and molecular mechanisms underlying the observed effects were not explored in detail. Future research should include a broader range of biomarkers and mechanistic studies to elucidate the pathways involved. Finally, the exact dosage and formulation of RO were based on previous animal studies, and optimal dosing for humans remains to be determined. These limitations highlight the need for further research to validate and expand upon the findings presented in this study.\u003c/p\u003e \u003cp\u003eAs described previously, the LF component of HRV is associated with sympathetic autonomic modulation, while the HF component is related to vagal modulation [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e, \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e, \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e]. In evaluating LF in the FD, both LF (ms\u0026sup2;) and LF (nu) showed no differences between the experimental groups; however, there was a noticeable trend towards a reduction in LF (nu) in the RO TSHR group. Additionally, when analyzing HF (ms\u0026sup2;), an improvement in vagal modulation was observed in the RO TSHR group, consistent with the findings related to parasympathetic tone, SD, variance, and RMSSD in this study. Ultimately, no differences were observed in the LF/HF ratio.\u003c/p\u003e \u003cp\u003eAutonomic modulation was also assessed using symbolic analysis, in which an increase at 0V and a decrease at 2V were observed in SHR, indicating sympathetic dominance and reduced parasympathetic modulation, respectively. These findings align with other studies showing autonomic imbalance in SHR [\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e]. Similarly, to the findings in the FD, only the RO\u0026thinsp;+\u0026thinsp;PE group was able to promote an improvement in the autonomic nervous system by reducing 0V and increasing 2V.\u003c/p\u003e \u003cp\u003eTherefore, the results of the analysis of autonomic variability via linear and non-linear methods, as well as parasympathetic tone, illustrate that the combination of RO with PE was the only treatment capable of inducing hypotension accompanied by an improvement in the cardiac autonomic nervous system. This indicates the need to explore other exercise protocols in combination with RO to further investigate the potential association of this dual intervention in controlling systemic arterial hypertension.\u003c/p\u003e \u003cp\u003eResults also showed attenuated baroreflex in SHR (after phenylephrine administration) compared to Wistar, as expected. However, none of the treatments were effective in improving baroreflex sensitivity. Furthermore, other studies have reported improvements in this parameter, since it was possible to observe enhancement of baroreflex in animals undergoing aerobic exercise in combination with antihypertensive drugs [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e, \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e]. Given these findings, we hypothesize that the exercise intervention chosen for this study was not effective in improving this variable.\u003c/p\u003e \u003cp\u003eAnalyzing the IHR and other studies that have addressed this parameter, it is worth mentioning that the decrease in IHR in SHR animals is consistent with sympathetic hyperactivity, along with electrical and structural remodeling of individual cells in the cardiac atrial pacemaker [\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e, \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e]. Moreover, the interventions were not effective in improving IHR. The main findings of the current study are illustrated in Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003e.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eIn summary, this was the first study to evaluate the hypotensive activity of chronic treatment with the monoterpene Rose Oxide in combination with physical exercise, showing that RO was able to reduce blood pressure levels in SHR both in TCP and PAP, along with improvements observed in the autonomic control of blood pressure through analysis of HRV and autonomic tone. RO is, therefore, a potential antihypertensive tool. Adjustments of the physical training variables and dosage could lead to promising results.\u003cb\u003e\u0026zwnj;\u003c/b\u003e\u003c/p\u003e"},{"header":"Declarations","content":"\u003ch2\u003eFunding\u003c/h2\u003e \u003cp\u003eThis study was supported by grants CNPq 409109/2018-5, 306987/2021-0, 423999/2021-4 and FAPEPI 00110.000235/2022-78.\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eJuliana A. da Silva, Samuel de S. P. Ara\u0026uacute;jo, Ana Fl\u0026aacute;via M. da Silva, Jos\u0026eacute; Guilherme V. de Assun\u0026ccedil;\u0026atilde;o, P\u0026acirc;mela de S. Santos, Jos\u0026eacute; L. P. J\u0026uacute;nior, Carlos Eduardo S. Reis, Liana de M. Santana, Regina G. Silva, Ariell A. de Oliveira, Francisca V. de Sousa Nunes: Performed the experimental and treatment protocol. Juliana A. da Silva, Samuel de S. P. Ara\u0026uacute;jo and Jo\u0026atilde;o Paulo J. Sabino wrote the main manuscript.Aldeidia P. de Oliveira, Dami\u0026atilde;o P. de Sousa, Renato N. Soriano, Luiz G. S. Branco, Helio C. Salgado, Jo\u0026atilde;o Paulo J. Sabino Analysis of resultsAll authors reviewed the manuscript\u003c/p\u003e\u003ch2\u003eAcknowledgments\u003c/h2\u003e \u003cp\u003eThis study was supported by grants CNPq 409109/2018-5, 306987/2021-0, 423999/2021-4 and FAPEPI 00110.000235/2022-78.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAL GHORANI, H. et al. Arterial hypertension \u0026ndash; Clinical trials update 2021. 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Journal of Cardiovascular Pharmacology, v. 74, n. 6, p. 542\u0026ndash;548, 2019. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1097/FJC.0000000000000746\u003c/span\u003e\u003cspan address=\"10.1097/FJC.0000000000000746\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"pflugers-archiv-european-journal-of-physiology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"paej","sideBox":"Learn more about [Pflügers Archiv - European Journal of Physiology](http://link.springer.com/journal/424)","snPcode":"424","submissionUrl":"https://submission.nature.com/new-submission/424/3","title":"Pflügers Archiv - European Journal of Physiology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Autonomic Control, Rose Oxide, Hypertension, SHR","lastPublishedDoi":"10.21203/rs.3.rs-4939277/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4939277/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eWith the alarming rise in cases of arterial hypertension worldwide, there is an urgent need to develop combined therapies to mitigate this scenario. Rose Oxide (RO), a monoterpene with anti-inflammatory and hypotensive properties, emerges as an alternative. The present study is the first to evaluate the effect of RO administered chronically and combined with physical exercise (swimming) since both have been reported to have beneficial impacts on hypertension. Male SHR and Wistar rats (aged 12 weeks) received RO for 34 consecutive days (orally; 100 mg/kg). The progression of systolic blood pressure (SBP) was monitored through tail-cuff plethysmography. Twenty-four hours before the end of the treatment, the animals were anesthetized, and the femoral artery and vein were cannulated to record the pulsatile arterial pressure and to administer drugs, respectively. Hemodynamic and autonomic parameters and baroreflex sensitivity and intrinsic heart rate (IHR) were evaluated. Treatment with RO, administered alone or combined with exercise, reduced SBP and mean arterial pressure in SHR. The swimming protocol did not prevent increases in BP, but when combined with RO, it improved autonomic control, assessed through heart rate variability and parasympathetic tone. IHR was attenuated in SHR, and none of the treatments reversed this response. Therefore, combining RO with physical exercise may enhance their antihypertensive effects, improving autonomic function, reducing oxidative stress and inflammation, providing synergistic cardiovascular benefits, improving metabolic health, promoting a comprehensive lifestyle intervention, and potentially allowing for reduced medication dosages. This multifaceted approach could offer a more effective and sustainable strategy for managing hypertension.\u003c/p\u003e","manuscriptTitle":"Chronic Rose Oxide and Exercise Synergistically Modulate Cardiovascular and Autonomic Functions in Hypertensive Rats","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-10-15 09:46:19","doi":"10.21203/rs.3.rs-4939277/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2024-09-20T19:02:22+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-09-20T15:46:17+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"276158447367620458690247440347174326918","date":"2024-09-11T16:10:05+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"311609118197903650433836145958477191880","date":"2024-09-10T19:53:06+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-09-02T09:01:35+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"252357178534450486904567163232199115900","date":"2024-08-27T18:36:07+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2024-08-26T15:57:20+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-08-22T00:48:52+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2024-08-22T00:48:14+00:00","index":"","fulltext":""},{"type":"submitted","content":"Pflügers Archiv - European Journal of Physiology","date":"2024-08-19T14:19:02+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"pflugers-archiv-european-journal-of-physiology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"paej","sideBox":"Learn more about [Pflügers Archiv - European Journal of Physiology](http://link.springer.com/journal/424)","snPcode":"424","submissionUrl":"https://submission.nature.com/new-submission/424/3","title":"Pflügers Archiv - European Journal of Physiology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"823e25ee-dcb3-4545-a580-bafa0fa1705b","owner":[],"postedDate":"October 15th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2024-11-04T16:21:12+00:00","versionOfRecord":{"articleIdentity":"rs-4939277","link":"https://doi.org/10.1007/s00424-024-03035-7","journal":{"identity":"pflugers-archiv-european-journal-of-physiology","isVorOnly":false,"title":"Pflügers Archiv - European Journal of Physiology"},"publishedOn":"2024-10-30 15:56:53","publishedOnDateReadable":"October 30th, 2024"},"versionCreatedAt":"2024-10-15 09:46:19","video":"","vorDoi":"10.1007/s00424-024-03035-7","vorDoiUrl":"https://doi.org/10.1007/s00424-024-03035-7","workflowStages":[]},"version":"v1","identity":"rs-4939277","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4939277","identity":"rs-4939277","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}