Expression Profiles of Circular RNA in Aortic Vascular Tissues of Spontaneously Hypertensive Rats

Background: Circular RNAs (circRNAs), as a kind of endogenous non-coding RNA, have been implicated in ischemic heart diseases and vascular diseases. Based on theirs high stability with a closed loop structure, circRNAs function as a sponge and bind specific miRNAs to exert inhibitory effects in heart and vasculature, thereby regulating their target gene and protein expression, via competitive endogenous RNA (ceRNA) mechanism. However, the exact roles and underlying mechanisms of circRNAs in hypertension and related cardiovascular diseases remain largely unknown.Methods and Results: High-throughput RNA sequencing (RNA-seq) was used to analyze the differentially expressed (DE) circRNAs in aortic vascular tissues of spontaneously hypertensive rats (SHR). Compared with the Wistar-Kyoto (WKY) rats, there were marked increases in the levels of systolic blood pressure, diastolic blood pressure and mean blood pressure in SHR under awake conditions via the tail-cuff methodology. Totally, compared with WKY rats, 485 DE circRNAs were found in aortic vascular tissues of SHR with 279 up-regulated circRNAs and 206 down-regulated circRNAs. Furthermore, circRNA-target microRNAs (miRNAs) and the target messenger RNAs (mRNAs) of miRNAs were predicted by the miRanda and Targetscan softwares, respectively. Additionally, real-time RT-PCR analysis verified that downregulation of rno_circRNA_0009197, and upregulation of rno_circRNA_0005818, rno_circRNA_0005304, rno_circRNA_0005506, and rno_circRNA_0009301 were observed in aorta of SHR when compared with that of WKY rats. Then, the potential ceRNA regulatory mechanism was constructed via integrating 5 validated circRNAs, 31 predicted miRNAs, and 266 target mRNAs. More importantly, three hub genes (NOTCH1, FOXO3, and STAT3) were recognized according to PPI network and three promising circRNA-miRNA-mRNA regulatory axes were found in hypertensive rat aorta, including rno_circRNA_0005818/miR-615/NOTCH1, rno_circRNA_0009197/ miR-509-5p/FOXO3, and rno_circRNA_0005818/miR-10b-5p/STAT3, respectively.Conclusions: Our results demonstrated for the first time that circRNAs are expressed aberrantly in aortic vascular tissues of hypertensive rats and may serve as a sponge linking with relevant miRNAs participating in pathogenesis of hypertension and related ischemic heart diseases via the circRNA-miRNA-mRNA ceRNAnetwork mechanism.

Short-term effect of ligature-induced periodontitis on cardiovascular variability and inflammatory response in spontaneously hypertensive rats

Background We previously reported that periodontal disease (PD) induces high arterial pressure variability (APV) consistent with sympathetic overactivity and elicits myocardial inflammation in Balb/c mice. However, it is unknown whether PD can change APV and heart rate variability (HRV) in spontaneously hypertensive (SHR) and normotensive Wistar-Kyoto (WKY) rats. This study aimed to evaluate the hemodynamic level, HRV, and APV associating with myocardial inflammation and plasma concentrations of oxide nitric (NO) in SHR and WKY rats with PD. Methods Three weeks after bilateral ligation of the first mandibular molar, or Sham operation, the rats received catheters into the femoral artery and had their arterial pressure (AP) recorded the following day. Subsequently, plasma, heart, and jaw were collected. The NO was quantified by the chemiluminescence method in plasma, and the myocardial IL-1β concentrations were evaluated by ELISA. In the jaw was evaluated linear alveolar bone loss induced by PD. Results The linear alveolar bone loss in jaws of SHR with PD was higher than in all other groups. AP and heart rate were higher in SHR than in their WKY counterparts. SHR with PD showed lower AP than control SHR. HRV and APV were different between SHR and WKY rats; however, no differences in these parameters were found between the animals with PD and their control counterparts. Plasma NO and myocardial IL-1β concentrations were higher in SHR with PD as compared to control WKY. A significant correlation was found between linear alveolar bone loss and plasma NO and myocardial IL-1β concentrations. Conclusion Our results demonstrated that short-term PD lowered the AP in SHR, which might be due to the higher levels of plasma NO. Even though PD did not affect either HRV or APV, it did induce myocardial inflammation, which can determine cardiovascular dysfunction in long-term PD.

Insufficiency of ventral hippocampus to medial prefrontal cortex transmission explains antidepressant non-response

Background: There is extensive evidence that antidepressant drugs restore normal brain function by repairing damage to ventral hippocampus (vHPC) and medial prefrontal cortex (mPFC). While the damage is more extensive in hippocampus, the evidence of treatments, such as deep brain stimulation, suggests that functional changes in prefrontal cortex may be more critical. We hypothesized that antidepressant non-response may result from an insufficiency of transmission from vHPC to mPFC. Method: Antidepressant non-responsive Wistar Kyoto (WKY) rats were subjected to chronic mild stress (CMS), then treated with chronic daily administration of the antidepressant drug venlafaxine (VEN) and/or repeated weekly optogenetic stimulation (OGS) of afferents to mPFC originating from vHPC or dorsal HPC (dHPC). Results: As in many previous studies, CMS decreased sucrose intake, open-arm entries on the elevated plus maze (EPM), and novel object recognition (NOR). Neither VEN nor vHPC–mPFC OGS alone was effective in reversing the effects of CMS, but the combination of chronic VEN and repeated OGS restored normal behaviour on all three measures. dHPC–mPFC OGS restored normal behaviour in the EPM and NOR test irrespective of concomitant VEN treatment, and had no effect on sucrose intake. Conclusions: The synergism between VEN and vHPC–mPFC OGS supports the hypothesis that the antidepressant non-responsiveness of WKY rats results from a failure of antidepressant treatment fully to restore transmission in the vHPC–mPFC pathway.

Differential Antihypertensive Effects of Oral Doses of Acetylcholine between Spontaneously Hypertensive Rats and Normotensive Rats

Acetylcholine (ACh) is a novel antihypertensive food component. Here, we demonstrate the differential effects of oral ACh on high and normal blood pressure in rats. Spontaneously hypertensive rats (SHRs) and Wistar–Kyoto (WKY) rats were administered ACh orally. The blood pressure and heart rate of SHRs were significantly lowered with ACh doses of 10−5 and 10−3 mol/kg body weight (b.w.), and the urinary catecholamine levels were significantly decreased with 10−3 mol/kg b.w. In contrast, oral ACh administration had no effect on WKY rats. This difference was likely caused by differences in sympathetic nervous activity and the baroreflex between strains. Comparison of gene sequences between the two strains revealed Chga mutations, suggesting that changes in the expression of chromogranin A might be involved in the baroreflex in SHRs. Oral ACh had an antihypertensive effect under hypertension but not normotension, indicating that this may be used safely to prevent hypertension.

Circumventricular Organ Apelin Receptor Knockdown Decreases Blood Pressure and Sympathetic Drive Responses in the Spontaneously Hypertensive Rat

The central site(s) mediating the cardiovascular actions of the apelin-apelin receptor (APJ) system remains a major question. We hypothesized that the sensory circumventricular organs (CVOs), interfacing between the circulation and deeper brain structures, are sites where circulating apelin acts as a signal in the central nervous system to decrease blood pressure (BP). We show that APJ gene (aplnr) expression was elevated in the CVOs of spontaneously hypertensive rats (SHRs) compared to normotensive Wistar Kyoto (WKY) controls, and that there was a greater mean arterial BP (MABP) decrease following microinjection of [Pyr1]apelin-13 to the CVOs of SHRs compared to WKY rats. Lentiviral APJ-specific-shRNA (LV-APJ-shRNA) was used to knockdown aplnr expression, both collectively in three CVOs and discretely in individual CVOs, of rats implanted with radiotelemeters to measure arterial pressure. LV-APJ-shRNA-injection decreased aplnr expression in the CVOs and abolished MABP responses to microinjection of [Pyr1]apelin-13. Chronic knockdown of aplnr in any of the CVOs, collectively or individually, did not affect basal MABP in SHR or WKY rats. Moreover, knockdown of aplnr in any of the CVOs individually did not affect the depressor response to systemic [Pyr1]apelin-13. By contrast, multiple knockdown of aplnr in the three CVOs reduced acute cardiovascular responses to peripheral [Pyr1]apelin-13 administration in SHR but not WKY rats. These results suggest that endogenous APJ activity in the CVOs has no effect on basal BP but that functional APJ in the CVOs is required for an intact cardiovascular response to peripherally administered apelin in the SHR.

Effect of the level of magnesium in drinking water on the state of the cardiovascular system of spontaneous hypertensive rats

INTRODUCTION. Magnesium is the second most common intracellular cation, is a cofactor for more than 300 enzymes, affects the functional state of the cardiovascular system through various mechanisms, in particular, through the action on the smooth muscle cells of the vessels, modulation of the renin-angiotensin-aldosterone system, regulation of sodium and calcium homeostasis. Therefore, maintaining a normal level of magnesium in the blood is an urgent task, and the consumption of drinking water enriched with magnesium can be considered as a method of correcting an insufficient intake of exogenous magnesium.THE AIM. The purpose of the study was determined – to evaluate the effect of drinking water with different contents of magnesium ions and a complex of magnesium with calcium on the state of the cardiovascular system of rats with genetically determined arterial hypertension. MATERIALS AND METHODS. From 6–7 weeks of age, male SHR rats received drinking water of various compositions for two months: in the first group (hCа+Mg) – with increased content of calcium and magnesium (120 mg/l Ca2+ and 45 mg/l Mg2+), the second (nCа+Mg) – drinking water normalized by mineral composition (60 mg/l Ca2+ and 25 mg/l Mg2+), in the third (hMg) – enriched Mg2+ (45 mg/l), the fourth (control) control group – St. Petersburg tap water with a low mineral content (8 mg/l Ca2+ and 3 mg/l Mg2+). WKY rats were divided into 2 groups: one group (hMg) received water enriched with Mg2+ (45 mg/l), the control WKY (control) group received water with a low mineral content (8 mg/l Ca2+ and 3 mg/l Mg2+). After 2 months, the blood pressure of rats on the tail was measured by the cuff method, the level of urea, cholesterol, total calcium, and albumin in the blood serum was analyzed, left ventricular mass index (LVMI) and myocardial mass index (MMI) were calculated. The spontaneous contractile activity of the portal vein (PV) was recorded by myography (in vitro) in isometric mode. The following were analyzed: frequency, total and maximum amplitude of phase-tonic contractions, the area under the contraction curve in 1 min, which characterizes the work performed by the vein. %). RESULTS. Enrichment of drinking water with Ca2+ and Mg2+ had a more pronounced antihypertensive effect in SHR rats compared with the hMg2+ group. In WKY rats, magnesium enrichment of water did not affect blood pressure. Modification of the mineral composition of drinking water did not affect MMI and LVMI in both SHR and WKY rats. Interlinear differences were found in the contractile activity of PV in control rats (the amplitude of PV contractions in SHR rats was greater than WKY. Consumption of water enriched with minerals decreased the amplitude of PV contractions, the largest decrease was in the hMg2+ group (in SHR, 2.6 times, in WKY, 1.5 times as compared to the control of the corresponding line). The value of the work performed by the PV in the control SHR rats was greater than in the control rats of the WKY line, and the enrichment of water with magnesium caused a decrease in the work performed by the PV only in rats SHR lines (by 55.6 %), but not for WKY. CONCLUSIONS. In rats, the consumption of drinking water enriched only with magnesium has an antihypertensive effect; however, it suppresses the spontaneous contractile activity of PV. It is advisable to use a complex of magnesium with calcium, which lowers blood pressure, but maintains an adequate level of contractile activity of the PV.

Validation of the Wistar-kyoto Rat Kept in Solitary Housing as an Animal Model for Endogenous Depression Using Voxel-based Morphometry

Major depressive disorder is a common psychiatric condition that is often resistant to medication. The Wistar-Kyoto (WKY) rat has been suggested as an animal model of endogenous depression; however, it is challenging to translate results obtained in animal models into humans. Solitary housing is a mild stress paradigm that could simulate the environment of depressive patients with limited social activity due to symptoms. We used voxel-based morphometry to directly compare the solitary-housed WKY rat model with data from previous human studies, and validated our results with behavioural studies and correlation analyses. Atrophy in WKY rats was detected in the ventral hippocampus, caudate putamen, lateral septum, cerebellar vermis, and cerebellar nuclei (p < 0.05, corrected for family-wise error rate). Further, locomotor behaviour was negatively correlated with hippocampal atrophy and positively correlated with atrophy of the cerebellar vermis. The regions of brain atrophy validate WKY rats as an animal model for endogenous depression and can aid the translation of study results to humans. Our study also reveals the possibility of a cerebellar contribution to depression.

Long-term inhibition of UCHL1 decreases hypertension and retinopathy in spontaneously hypertensive rats

Objective To investigate the role of the deubiquitinase ubiquitin C-terminal hydrolase L1 (UCHL1) in hypertension and retinopathy in the spontaneously hypertensive rat (SHR). Methods Wistar–Kyoto (WKY) rats and SHRs were administered the UCHL1 inhibitor LDN57444 (20 μg/kg/day) for 4 months. Pathological changes were detected with hematoxylin and eosin, immunofluorescence, and dihydroethidium staining. The mRNA and protein expression of UCHL1 were examined by real-time PCR and immunoblotting analysis. Results At 6 months of age, SHRs showed significantly increased mRNA and protein levels of UCHL1 in the retina compared with WKY rats. Moreover, SHRs exhibited significantly increased central retinal thickness, inflammation, and reactive oxygen species production compared with WKY rats, and these effects were markedly attenuated by systemic administration of the UCHL1 inhibitor LDN57444. The beneficial effects of LDN57444 were possibly associated with reduced blood pressure and the inactivation of several signaling pathways. Conclusion UCHL1 is involved in hypertension and retinopathy in SHRs, suggesting that UCHL1 may be used as a potential therapeutic target for treating hypertensive retinopathy.

Fructose Intake Impairs the Synergistic Vasomotor Manifestation of Nitric Oxide and Hydrogen Sulfide in Rat Aorta

In this study, we evaluated the effect of eight weeks of administration of 10% fructose solution to adult Wistar Kyoto (WKY) rats on systolic blood pressure (SBP), plasma and biometric parameters, vasoactive properties of the thoracic aorta (TA), NO synthase (NOS) activity, and the expression of enzymes producing NO and H2S. Eight weeks of fructose administration did not affect SBP, glycaemia, or the plasma levels of total cholesterol or low-density and high-density lipoprotein; however, it significantly increased the plasma levels of γ-glutamyl transferase and alanine transaminase. Chronic fructose intake deteriorated endothelium-dependent vasorelaxation (EDVR) and increased the sensitivity of adrenergic receptors to noradrenaline. Acute NOS inhibition evoked a reduction in EDVR that was similar between groups; however, it increased adrenergic contraction more in fructose-fed rats. CSE inhibition decreased EDVR in WKY but not in fructose-fed rats. The application of a H2S scavenger evoked a reduction in the EDVR in WKY rats and normalized the sensitivity of adrenergic receptors in rats treated with fructose. Fructose intake did not change NOS activity but reduced the expression of eNOS and CBS in the TA and CSE and CBS in the left ventricle. Based on our results, we could assume that the impaired vascular function induced by increased fructose intake was probably not directly associated with a decreased production of NO, but rather with impairment of the NO–H2S interaction and its manifestation in vasoactive responses.

Model Corrected Blood Input Function to Compute Cerebral FDG Uptake Rates From Dynamic Total-Body PET Images of Rats in vivo

Recently, we developed a three-compartment dual-output model that incorporates spillover (SP) and partial volume (PV) corrections to simultaneously estimate the kinetic parameters and model-corrected blood input function (MCIF) from dynamic 2-[18F] fluoro-2-deoxy-D-glucose positron emission tomography (FDG PET) images of mouse heart in vivo. In this study, we further optimized this model and utilized the estimated MCIF to compute cerebral FDG uptake rates, Ki, from dynamic total-body FDG PET images of control Wistar–Kyoto (WKY) rats and compared to those derived from arterial blood sampling in vivo. Dynamic FDG PET scans of WKY rats (n = 5), fasted for 6 h, were performed using the Albira Si Trimodal PET/SPECT/CT imager for 60 min. Arterial blood samples were collected for the entire imaging duration and then fitted to a seven-parameter function. The 60-min list mode PET data, corrected for attenuation, scatter, randoms, and decay, were reconstructed into 23 time bins. A 15-parameter dual-output model with SP and PV corrections was optimized with two cost functions to compute MCIF. A four-parameter compartment model was then used to compute cerebral Ki. The computed area under the curve (AUC) and Ki were compared to that derived from arterial blood samples. Experimental and computed AUCs were 1,893.53 ± 195.39 kBq min/cc and 1,792.65 ± 155.84 kBq min/cc, respectively (p = 0.76). Bland–Altman analysis of experimental vs. computed Ki for 35 cerebral regions in WKY rats revealed a mean difference of 0.0029 min−1 (~13.5%). Direct (AUC) and indirect (Ki) comparisons of model computations with arterial blood sampling were performed in WKY rats. AUC and the downstream cerebral FDG uptake rates compared well with that obtained using arterial blood samples. Experimental vs. computed cerebral Ki for the four super regions including cerebellum, frontal cortex, hippocampus, and striatum indicated no significant differences.