Renal Tissue Norepinephrine Research Articles

Optimal Strategy for HIFU-Based Renal Sympathetic Denervation in Canines.

Background: The association between the treatment efficacy and safety of high-intensity focused ultrasound (HIFU)-based renal sympathetic denervation (RDN) and the acoustic energy dose applied has not been fully studied and may provide important understanding of the mechanism that led to failure of the WAVE IV trial. The objective of this study was to externally deliver different HIFU doses to canines for RDN treatment and to investigate the optimal energy dose for HIFU-based RDN.Methods: Thirty canines were divided into five RDN groups according to dose of acoustic energy applied, and a sham control group that consisted of four canines was used for comparisons. All animals in the RDN groups underwent the RDN procedure with different acoustic energy doses, while in the sham control group, renal arteries were harvested without being subjected to acoustic energy delivery and were imaged using color Doppler flow imaging (CDFI). Blood pressure (BP) was recorded, and blood samples were collected before the RDN procedure and at 28 days after the RDN procedure. Histological examinations and measurement of renal tissue norepinephrine concentration were performed in all retrieved samples.Results: Suppression of BP was significant in the 300 W (15.17/8.33 ± 1.47/1.21 mmHg), 250 W (14.67/9.33 ± 1.21/1.37 mmHg), and 200 W (13.17/9.17 ± 2.32/1.84 mmHg) groups. Semiquantitative histological assessment of periarterial nerves around the kidney revealed that target nerves in the 300 W (9.77 ± 0.63), 250 W (9.42 ± 0.67), and 200 W (9.58 ± 0.54) groups had the highest nerve injury scores, followed by the 150 W group (5.29 ± 0.62). Furthermore, decreased renal tissue norepinephrine concentration, together with decreased expression of tyrosine hydroxylase in the 300, 250, and 200 W groups demonstrated effective sympathetic depression following sufficient acoustic energy deposition. However, the renal artery injury score in the 300 W group (0.93 ± 0.13) was significantly higher than in the other groups (p < 0.001).Conclusion: This study provides evidence that RDN effectiveness is based on the energy dose delivered and that 200–250 W is effective and safe in normal-sized canines.

Renal denervation improves vascular endothelial dysfunction by inducing autophagy via AMPK/mTOR signaling activation in a rat model of type 2 diabetes mellitus with insulin resistance.

Recent clinical and animal studies have shown that renal denervation (RDN) improves insulin sensitivity and endothelial dysfunction. However, the specific mechanism remains incompletely understood. The purpose of this study is to investigate the effects of RDN on endothelial dysfunction of type 2 diabetes mellitus (T2DM) rat models with insulin resistance and to explore the underlying molecular mechanisms. Male Sprague-Dawley rats were fed with or without high-fat diet allocated in different groups, combined with low-dose streptozotocin which induces a rat model to develop T2DM with insulin resistance. RDN was conducted 1week after the rat models fully developed T2DM. The animals were sub-divided into four groups randomly: control group (CON, n = 6), diabetic group (T2DM, n = 6), diabetic with sham surgery group (Sham, n = 6) and diabetic with RDN group (RDN, n = 6). Rats in all groups were studied at baseline, both preoperatively and 4weeks after RDN, respectively. Western blot was used to detect the expression of angiotensin-converting enzyme 2 (ACE2) protein and the expression of autophagy-related proteins Beclin1, LC3 and p62 and autophagy signaling pathway AMPK/mTOR proteins and apoptosis-related protein caspase-3 in the aorta endothelial cells. In addition, the effects of ACE2 on autophagy of human umbilical vein insulin resistance endothelial cell culture in vitro were also studied. RDN decreased plasma and renal tissue norepinephrine levels. The Von Willebrand factor level was also decreased, while the plasma level of nitric oxide (NO) was significantly increased after RDN. Compared with the T2DM group and the Sham group, the endothelium-dependent and endothelium-independent diastolic function of the RDN group was improved significantly, the expression of Beclin1, LC3, ACE2 and eNOS proteins was higher, and the level of p62 protein was decreased. Furthermore, we found that RDN can activate the expression of p-AMPK and inhibit the expression of p-mTOR. In cell culture experiment, ACE2 activated p-AMPK and inhibited p-mTOR, thus promoting autophagy. RDN may not only increase the expression of ACE2 in the vascular endothelium, but also can via ACE2 activate p-AMPK and inhibit p-mTOR, thus promoting autophagy and improving endothelial dysfunction.

1197The development and validation of a novel transcatheter microwave renal denervation system

Abstract Background Clinical studies of transcatheter radiofrequency renal denervation for treating hypertension have been hampered by the lack of consistent denervation efficacy. Microwave energy is well suited to renal denervation due to its capacity to spare vascular structures due to cooling from adjacent blood flow while enabling deep perivascular heating. Purpose We aimed to: 1) develop a transcatheter microwave system capable of safely delivering deep and circumferential perivascular renal nerve ablation, and 2) demonstrate the feasibility, short-term efficacy and safety of transcatheter microwave renal denervation. Method A novel 7F transcatheter microwave denervation system was designed, built, and iteratively prototyped in vitro and in 15 sheep. A histological grading system for microwave induced renal arterial and renal nerve injury was devised. The microwave denervation system was validated in an additional 9 sheep, which underwent unilateral renal denervation. Up to 2 microwave ablations were delivered to each artery with maximum power at 100–110W for 480s. Sheep were euthanised at 2–3 weeks post procedure. Gross microscopic histological examination as well as renal tissue norepinephrine content was analysed. Results Catheter deployment and ablation was successful in all 19 targeted vessel segments and ablation produced substantial circumferential perivascular injury; median ablation lesion area &gt;395 (IQR 251–437) mm2, depth 17.1 (IQR 15.8–18.4) mm, length 16 (IQR 12–20) mm, without collateral visceral injury. Limiting power to 100W minimised arterial injury, while maintaining a deep circumferential perivascular ablation. At microwave ablation sites, a total of 292 nerve fascicles were identified, median distance from the renal artery of 4.2mm (IQR 2.1–8.8mm), of which 249 (85%) had sustained thermal injury with 128/249 (51%) showing grade 3–4 (moderate to severe) injury. Microwave denervation reduced median functional sympathetic nerve surface area at the renal hilum on anti-tyrosine hydroxylase staining by 100% (IQR 87%-100%), p=0.0039, and median renal cortical norepinephrine content by 83% (IQR 76%–92%), p=0.0078, compared to the paired control kidney. Conclusion Transcatheter microwave ablation can produce deep circumferential perivascular ablations over a long segment of the renal artery without significant arterial or collateral visceral injury to provide effective renal denervation. Clinical translation may enable more consistent and complete transcatheter renal denervation and antihypertensive efficacy. Acknowledgement/Funding University of Sydney; Western Sydney Local health District; National Health and Medical Research Council of Australia; National Heart Foundation (Au)

Anatomy and neural remodeling of the renal sympathetic nerve in a canine model and patients with hypertension.

The role of renal sympathetic nerve (RSN) in hypertension should be better understood. We aimed to three-dimensionally reconstruct the renal nerves, and explore its anatomical and histochemical characteristics in hypertensive canine model and patients. Renal arteries with surrounding tissue were collected from canines and cadavers with or without hypertension. Serial renal artery hematoxylin-eosin sections were used for three-dimensional reconstruction, and morphological parameters were collected and analyzed. In hypertensive canines, the mean renal nerve number was 26.71 ± 5.68 versus 19.84 ± 5.68 in controls (P = 0.02), and the middle renal nerve volume was 5.31 ± 2.13 versus 2.60 ± 1.00 μl in controls (P = 0.01). Renal tissue norepinephrine concentrations, tyrosine hydroxylase and substance P immunoreactivity in RSN, and growth-associated protein 43 immunoreactivity in renal ganglion were significantly increased in hypertensive canines. In humans, the renal nerve was evenly distributed along the renal artery in a network pattern. The renal ganglion volume was 72.75 ± 33.43 in hypertensive patients versus 37.04 ± 23.95 μl in controls (P = 0.029) and the mean neuronal size in renal ganglion was 1187.3 ± 219.9 μm in patients versus 714.8 ± 142.7 μm in controls (P = 0.002). Tyrosine hydroxylase immunoreactivity in the RSN was 0.153 ± 0.014 in patients versus 0.104 ± 0.019 in controls (P = 0.013). Growth-associated protein 43 immunoreactivity in the renal ganglion was 86 612.8 ± 14 642.0 in patients versus 33 469.8 ± 15 666.8 μm/mm in controls (P < 0.001). Our study suggests that RSN and renal ganglion histological remodeling occurs in individuals with hypertension and the distal segment or branches of renal artery might be a promising therapeutic target for RSN modulation therapy.

Anti-hypertensive effect of radiofrequency renal denervation in spontaneously hypertensive rats

AimsWe aimed to investigate the anti-hypertensive effect of radiofrequency (RF) renal denervation (RDN) in an animal model of hypertension. Materials and methodsRF energy was delivered to bilateral renal arteries through a 2Fr catheter with opening abdomen in 8 spontaneously hypertensive rats (SHRs) and 8 Wistar–Kyoto rats (WKYs). Sham operation was performed in other 8 SHRs and 8 WKYs. Blood pressure (BP), heart rate (HR), and urinary norepinephrine excretion were followed up for 3months. Plasma and renal tissue concentrations of norepinephrine and plasma renin activity were measured 3months after the procedure. The RDN was confirmed by a decrease in renal tissue norepinephrine. Key findingsRF-RDN restrained a spontaneous rise in systolic BP (46±12% increase from 158±8 to 230±14mmHg vs. 21±18% increase from 165±9 to 197±20mmHg, p=0.01) and diastolic BP (55±27% increase from 117±9 to 179±23mmHg vs. 28±13% increase from 120±7 to 154±13mmHg, p=0.04) in SHRs; however, WKYs were not affected. Although there were no changes in HR and systemic norepinephrine, the renal tissue norepinephrine was decreased by RF-RDN in both SHR (302±41 vs. 159±44ng/g kidney, p<0.01) and WKY (203±33 vs. 145±26ng/g kidney, p=0.01). Plasma renin activity was reduced by the RF-RDN only in SHR (35.3±9.5 vs. 21.4±8.6ng/mL/h, p<0.01). SignificanceRF-RDN demonstrated an anti-hypertensive effect with a reduction of renal tissue norepinephrine and plasma renin activity in SHR.

Radiofrequency renal nerve denervation decreases blood pressure, sympathetic activity and renal norepinephrine content in spontaneously hypertensive rats (875.16)

Radiofrequency renal denervation (RF‐RDN) decreases blood pressure (BP) in patients with drug‐resistant hypertension. This study investigated the effects of RF‐RDN on sympathetic activity and BP (Systolic, SBP; Diastolic, DBP) in SHR. Methods Nineteen‐week old male SHR were instrumented with telemetry probes for chronic measurement of BP and BP variability (BPV). After 1‐week, control BP was measured in rats for 3 days. The next day SHR (n=6/group) were anesthetized and randomly received either bilateral Sham‐RDN or RF‐RDN of the renal arteries (Biosense Webster Stockert 70 generator and RF‐probe). BP was then measured in SHR for 8 weeks. Results Following RF‐RDN, SBP/DBP was significantly decreased the day after (153±8/102±4 mmHg) and throughout the post‐RDN study (4‐week SBP/DBP, 173±7/120±5; 8‐weeks, 171±6/116±4 mmHg) compared to pre‐RDN control levels (SBP/DBP, 187±9/130±5 mmHg). RF‐RDN significantly decreased the low frequency component of BPV in SHR with the peak attenuation observed at 8 weeks (LF/HF 2.2±0.2 vs 0.9±0.2); this correlated with a blunting of the hypotensive response to i.p. chlorisondamine in RF‐RDN rats. Finally, at the end of week 8, kidney norepinephrine (NE) levels in RF‐RDN rats (left, 10±10; right, 0.0±0.0 ng/g kidney) were dramatically decreased compared with levels from sham SHR (left, 130±10; right, 110±10 ng/g kidney). Conclusions These data demonstrate that in hypertensive SHR, bilateral RF‐RDN of the renal arteries decreases BP in part by chronic inhibition of sympathetic neural pathways and a decrease in renal tissue NE levels.Grant Funding Source: Biosense Webster grant IIS‐175 to DRK and FS; NIH NIGMS P20 GM103514 to DK

Effects of renal denervation on the development of post-myocardial infarction heart failure and cardiac autonomic nervous system in rats

Prior studies indicated that radiofrequency renal denervation (RD)had beneficial effects on post-myocardial infarction (MI) heart failure(HF) in rats[1–3]. In this studywe aimedtoassessits effects on cardiacautonomic nervous system (CANS) which might be one of the mostimportant mechanisms of RD's therapeutic effect on post-MI HF anddetermine the best timing for RD.One hundred Wistar rats were randomly assigned intofiveexperimental groups: MI group (n = 20), RD group (n = 20),MI-1d + RD group (RD performed one day post-MI, n = 20),MI-4w + RD group (RD performed four weeks post-MI, n = 20),andNgroup(controlgroup, n = 20).MIwasproducedthroughligationof the anterior descending artery. RD was performed through strippingof the renal nerves. The experimental design and implementation wereconducted in accordance with animal welfare guidelines.Eight weeks post-MI, significant improvements were observed inboth MI-1d + RD and MI-4w + RD groups compared to the MIgroup, that include (1) improved left ventricular (LV) function andhemodynamics with increased water and sodium excretion;(2) decreased plasma and renal tissue norepinephrine levelswhile tissue norepinephrine content increased in myocardium;(3) increased β1-receptor in myocardium and improved heartratevariability;and(4)decreasedplasmarenin,angiotensinII,aldoste-rone,BNPandendothelinlevels.Moretherapeuticeffectswerefoundinthe MI-1d + RD group than the MI-4w + RD group (see Table 1 andFig. 1).Firstly, our study showed that RD attenuated the remodeling ofCANS and modulated its activities. RD leads to preservation of β1receptors content along with the β1 mRNA expression in non-infarcted cardiac tissue in this HF model (Fig. 1). This correlated withan improvement in heart function and cardiac remodeling. HRV is asensitive marker for the CANS [4]. RD led to a slower HR and higherSDNN in both intervention groups (See Table 1). The increase in SDNNindicates that cardiac sympathetic over-activation was suppressed andvagal activity was enhanced with the result of stabilizing cardiacelectrical activity, decreasing malignant arrhythmias, thereby reducingthe incidence of sudden death.Secondly, we found that RD blocked both peripheral and centralRAAS and sympathetic nervous system (SNS) at the same time. Andthis may answer the question how RD exerted effect on CANS. In ourstudy RD restores renin, angiotensin II, and aldosterone to near-normal levels. This not only explains the increase in sodium and waterexcretion, but also confirms that RD blocks renal RAAS via blockage ofthe efferent renal sympathetic nerves which is consistent with ourprevious study [2]. Studies have shown that angiotensin II not onlyactivates the central and peripheral SNS, function as part of a positivefeedback cycle between RAAS and the SNS, but also activates the localRAAS of other organs, notably the heart [5]. Our study also showedthat the plasma NE level in MI-1d + RD and MI-4w + RD groupsdecreased significantly, which reflects an overall reduction ofsympathetic activity, and especially represents reduction of centralsympathetic activity. This result maybe in part due to blockade of therenal sympathetic afferent pathway by RD. Furthermore, previousstudies have shown that during HF, the tissue NE levels of both theheart and kidney are decreased due to depletion, decreased reuptakeand increased spillover. The decrease of the tissue NE level causedfeedback activation of the central sympathetic system [6].Thus,increased tissue NE level may alleviate the activation of the centralsympathetic system, which was reflected by our data (Fig. 1).Many studies have shown that central RAAS stimulates the central

The anti-hypertensive effect of radiofrequency renal denervation with a reduction of renal tissue norepinephrine content in the spontaneously hypertensive rats

The anti-hypertensive effect of radiofrequency renal denervation with a reduction of renal tissue norepinephrine content in the spontaneously hypertensive rats

Cryo-treatment of the renal nerve to the clipped kidney in 2K1C Hypertensive rats: Hemodynamic and neural responses

R renal denervation (DNX) is gaining increasing interest as a modality for treating resistant hypertension. The two-kidney one-clip (2K1C) hypertensive rat is a model of renovascular hypertension. In addition to activation of the reninangiotensinsystem, 2K1C rats display elevated sympathetic activity. Earlier work reported that surgical renal denervation (DNX) decreases mean arterial pressure (MAP) in this model. Since radiofrequency ablation may be problematic in atherosclerotic renal artery disease, we hypothesized that cryo-treatment of the renal nerve to the clipped kidney of 2K-1C rats would decrease MAP, decrease contralateral renal sympathetic nerve activity (RSNA), and improve baroreflex sensitivity. Male Sprague-Dawley rats had sham (SC) or right renal artery clipping (2K1C) and hemodynamic monitoring by telemetry. They were pair fed a 0.4% NaCl diet. Six weeks later,2K1C rats were stratified to moderate (MAP≥140 mmHg) or mild (MAP<140 mmHg) hypertension, instrumentedand underwent sham-DNX or cryo-DNX. All rats were studied in the conscious state. MAP decreased by 16±5 mmHg in the 2K1C cryo-DNXmoderate hypertension group (P<0.05). Cryo-DNX exerted only a small depressor effect in mildly hypertensive 2K1C rats. In the moderately hypertensive 2K1C cryo-DNX rats, contralateral RSNA decreased from 12.2±1.5 to 3.1±3.5 μV.s (P<0.05), a value no different from SC rats 3.3 μV.s. Baroreflex sensitivity of both heart rate and RSNA increased in the cryo-DNX 2K1C rats. Renal tissue norepinephrine decreased by 73% in the DNX kidney; renal tissue Ang II was not altered. Approaches using cryo-treatment to ablate renal nerves may provide an adjunctive therapeutic modality in resistant renovascular hypertension.

Evidence for Sympathetic Origins of Hypertension in Juvenile Offspring of Obese Rats

Maternal obesity in rodents is associated with increased adiposity, impaired glucose tolerance, and hypertension in adult offspring. In this study we investigated the influence of maternal obesity in the rat on blood pressure and blood pressure regulatory pathways in juvenile and adult offspring. Obesity was induced before pregnancy in female Sprague-Dawley rats by feeding a highly palatable energy-dense diet. In juvenile animals (30 days of age), before the onset of obesity and hyperleptinemia, basal nighttime mean arterial pressure was significantly raised in the offspring of obese dams (OffOb) relative to offspring of controls (OffCon; mean arterial pressure, males: OffOb, 121.8+/-0.6 mm Hg versus OffCon, 115.0+/-0.5 mm Hg, n=6, P<0.01; females: OffOb, 125.4+/-0.4 mm Hg versus OffCon, 114.4+/-0.5 mm Hg, n=6, P<0.001), as was the mean arterial pressure response to restraint stress (P<0.01). The pressor response to a leptin challenge was enhanced in OffOb rats (Deltamean arterial pressure: OffOb, 9.7+/-0.8 mm Hg versus OffCon, 5.3+/-1.3 mm Hg; n=8; P<0.05). Renal tissue norepinephrine content (P<0.001) and renin expression (P<0.05) were markedly raised. Analysis of heart rate variability revealed an increased low:high frequency ratio in OffOb versus OffCon rats (P<0.05). At 90 days, hypertension in OffOb rats persisted and was abolished by alpha1- and beta-adrenergic blockade, and cardiovascular responses to phenylephrine or sodium nitroprusside indicated altered baroreceptor function. The exaggerated pressor response to leptin in OffOb rats was maintained. Hypertension in the offspring of obese rats may arise from persistent sympathoexcitatory hyperresponsiveness acquired in early stages of development.

Feeding-induced increase in urinary dopamine excretion is independent of renal innervation and sodium intake.

Food intake increases urinary dopamine excretion. To investigate the role of renal nerve activity in this renal response to feeding, we studied urinary dopamine excretion after chronic bilateral renal denervation (DNX, n = 8) in male Sprague-Dawley rats. Controls were sham operated (CON, n = 6). In a paired crossover design, animals were studied both in fed and fasted state. Time course measurements of renal tissue norepinephrine (NE) content in a third group of DNX rats showed a decrease by 90% not earlier than 18 h post-DNX, remaining to be reduced to this level until day 11. Renal NE content in DNX animals was measured at the end of the study to confirm complete denervation. In 24-h urine, collected from fed and fasted conscious CON and DNX rats in metabolic cages, concentrations of dopamine and sodium were measured during two different sodium intake regimens, i.e., tap water and 1 g/dl NaCl as drinking water. No influence of renal innervation on urinary dopamine could be observed in fed and fasted animals. The marked changes in urinary volume and sodium output due to the different sodium regimens were not paralleled by urinary dopamine excretion. However, urinary dopamine was increased 2.1- to 2.4-fold (P < 0.01) because of feeding in all groups, independent of sodium balance and renal innervation. We conclude that in conscious rats both basal urinary dopamine excretion and its marked increase in response to feeding are independent of sodium balance and of renal nerve activity.

Renal nerves in renal adaptation to dietary sodium restriction.

To assess the physiologic importance of the renal nerves in the renal mechanisms for the maintenance of body sodium balance, renal adaptation to normal and low sodium diet was evaluated in conscious Sprague-Dawley male rats before and 8 days after recovery from bilateral surgical-pharmacological renal denervation. Renal denervation was confirmed in every rat at the end of the study by absence of renal vasoconstriction to splanchnic nerve stimulation and loss of renal tissue norepinephrine content. Daily sodium balance, defined as the difference between dietary sodium intake and urinary sodium excretion, was positive with the normal sodium diet before and after bilateral renal denervation. Prior to bilateral renal denervation, changing to the low sodium diet was associated with a diminishingly negative sodium balance for 3 days that became progressively positive thereafter. After bilateral renal denervation, changing to the low sodium diet was associated with a continuous and progressively negative sodium balance. We conclude that intact renal innervation is required for normal renal sodium conservation and maintenance of body sodium balance during dietary sodium restriction.

Tissue catecholamines in obstructive nephropathy and acute uremia in the rat.

Previous studies have shown that the 24-h unilateral ureteral obstructed kidney (UUO) responds to acute renal denervation after relief of obstruction with an increase in glomerular filtration rate (GFR), renal plasma flow (RPF), natriuresis, and diuresis, while the 24-h bilateral ureteral obstructed kidney (BUO), undergoing postobstructive diuresis after relief of obstruction, shows no response to acute denervation. In the present study, renal tissue norepinephrine (NE) content and concentration were found to be markedly decreased in the BUO kidney suggesting partial renal denervation, while renal NE content was normal in the UUO kidney. The decrease in renal NE content in the BUO kidney was organ specific since myocardial NE was not affected, and was not due to acute uremia, since rats made uremic by 24 h of total intravenous urine reinfusion showed no change in renal NE content. Renal dopamine content was markedly increased in the UUO kidney, perhaps as a result of renal decarboxylation of dopa and failure of excretion, while renal and myocardial epinephrine content were increased in the BUO and urine reinfused groups, both of which were subjected to the stress of acute uremia. The decrease in renal NE content in the kidney undergoing postobstructive diuresis after relief of 24-h BUO probably indicates damage to sympathetic nerve terminals by high intrarenal pressure during BUO and suggests that partial renal denervation could contribute to the functional changes observed in this experimental model.