Search

Hypertension is a serious public health problem worldwide. MT-1207, chemically named 3-(4-(4-(1H-benzotriazole-1-yl)butyl)piperazine-1-yl) benzisothiazole hydrochloride, is a new chemical entity that has entered into clinical trial as antihypertensive agent in China. In this paper we report the pharmacological profile of MT-1207 regarding its acute, subacute, and long-term effects on hypertensive animal models, and its actions on isolated organs in vitro as well as its molecular targets. Blood pressure (BP) was measured in conscious animals; amlodipine was taken as a positive control drug. We showed that both single dose of MT-1207 (1.25−20 mg/kg, ig) in spontaneously hypertensive rats (SHR) and MT-1207 (0.25−6 mg/kg, ig) in two-kidney one-clip (2K1C) dogs dose-dependently decreased BP. MT-1207 quickly decreased BP within 5 min after administration; the hypotensive effect lasted for 8 and 12 h, respectively, in SHR and 2K1C dogs without reflex increase in heart rate. Multiple doses of MT-1207 (5 mg · kg(−1) · d(−1) in SHR; 2 mg · kg(−1) · d(−1) in 2K1C dogs, for 7 days) significantly decreased BP, slightly reduced heart rate, and both of them recovered after withdrawal. Long-term administration of MT-1207 (10 mg · kg(−1) · d(−1) for 4 months or more time) produced a stable BP reduction, improved baroreflex sensitivity, reduced renal and cardiovascular damage in SHR, and delayed stroke occurrence and death in stroke-prone SHR. In isolated rat aortic rings precontracted by adrenaline, KCl, noradrenaline or 5-hydroxytryptamine (5-HT), MT-1207 (10(−9)–10(−4) M) caused concentration-dependent relaxation. In a panel of enzyme activity or radioligand binding assays of 87 molecular targets, MT-1207 potently inhibited adrenergic α(1A), α(1B), α(1D), and 5-HT(2A) receptors with K(i)

MicroRNAs play pivotal roles in regulation of both innate and adaptive immune responses. In the present study, we investigated the effects of microRNA-124 (miR-124) on production of the pro-inflammatory cytokine TNF-α in lipopolysaccharide (LPS)-treated mouse macrophages. Mouse macrophage cell line RAW264.7 was stimulated with LPS (100 ng/mL). The levels of miR-124 and TNF-α mRNA were evaluated using q-PCR. ELISA and Western blotting were used to detect TNF-α protein level in cell supernatants and cells, respectively. 3′-UTR luciferase reporter assays were used to analyze the targets of miR-124. For in vivo experiments, mice were injected with LPS (30 mg/kg, ip). LPS stimulation significantly increased the mRNA level of miR-124 in RAW264.7 macrophages in vitro and mice in vivo. In RAW264.7 macrophages, knockdown of miR-124 with miR-124 inhibitor dose-dependently increased LPS-stimulated production of TNF-α protein and prolonged the half-life of TNF-α protein, but did not change TNF-α mRNA levels, whereas overexpression of miR-124 with miR-124 mimic produced the opposite effects. Furthermore, miR-124 was found to directly target two components of deubiquitinating enzymes: ubiquitin-specific proteases (USP) 2 and 14. Knockdown of USP2 or USP14 accelerated protein degradation of TNF-α, and abolished the effect of miR-124 on TNF-α protein stability. miR-124, targeting USP2 and USP14, negatively regulates LPS-induced TNF-α production in mouse macrophages, suggesting miR-124 as a new therapeutic target in inflammation-related diseases.

To evaluate the anti-effects of anisodamine and neostigmine in animal models of endotoxic and hemorrhagic shock. Kunming mice were injected with lipopolysaccharide (LPS 30 mg/kg, ip) to induce endotoxic shock. Anisodamine (12.5, 25, and 50 mg/kg, ip) and neostigmine (12.5, 25, and 50 μg/kg, ip) were administered immediately after LPS injection. Survival rate was monitored, and the serum levels of TNF-α and IL-1β were analyzed using ELISA assays. The effects of anisodamine and neostigmine were also examined in α7 nicotinic acetylcholine receptor (α7 nAChR) knockout mice with endotoxic shock and in Beagle dogs with hemorrhagic shock. In mice with experimental endotoxemia, combined administration of anisodamine and neostigmine significantly increased the survival rate and decreased the serum levels of inflammatory cytokines, as compared to those produced by either drug alone. The anti-shock effect of combined anisodamine and neostigmine was abolished in α7 nAChR knockout mice. On the other hand, intravenous injection of the combined anisodamine and neostigmine, or the selective α7 nAChR agonist PNU282987 exerted similar anti-shock effects in dogs with hemorrhagic shock. The results demonstrate that combined administration of anisodamine and neostigmine produces significant anti-shock effects, which involves activation of α7 nAChRs.

To investigate the synergism of low-doses of amlodipine and irbesartan on reduction of blood pressure variability (BPV), amelioration of baroreflex sensitivity (BRS) and organ protection in spontaneously hypertensive rats (SHR).The rats were administered amlodipine (1 mg·kg(-1)·d(-1)) alone, irbesartan (10 mg·kg(-1)·d(-1)) alone, or the combination of the two drugs for 4 months. The drugs were mixed into the rat chow. Blood pressure (BP) was continuously monitored in conscious animals. After the determination of BRS, the rats were killed for morphological evaluation of organ damages.The combination of low-dose irbesartan and amlodipine had statistically significant synergism on reduction of BP and BPV, amelioration of BRS and organ protection in SHR. Multiple regression analysis showed that the decrease in left ventricular hypertrophy was associated with the decrease in systolic BPV (r=0.665, P0.01); the decrease in aortic hypertrophy was associated with the increase in BRS (r=0.656, P0.01); and the amelioration in renal lesion was associated with the increase in BRS (r=0.763, P0.01) and the decrease in systolic BPV (r=0.706, P0.01).Long-term treatment with a combination of low-doses of amlodipine and irbesartan showed significant synergism on reduction of BP and BPV, restoration of BRS and organ protection in SHR. Besides BP reduction, the enhancement of BRS and reduction of BPV might contribute to the organ protection.

Stroke is the second most common cause of death and the most common cause of disability in developed countries. Stroke is a multi-factorial disease caused by a combination of environmental and genetic factors. Numerous epidemiologic studies have documented a significant genetic component in the occurrence of strokes. Genes encoding products involved in lipid metabolism, thrombosis, and inflammation are believed to be potential genetic factors for stroke. Although a large group of candidate genes have been studied, most of the epidemiological results are conflicting. Studies of stroke as a monogenic disease have made huge progress, and animal models serve as an indispensable tool to dissect the complex genetics of stroke. In the present review, we provide insight into the role of in vivo stroke models for the study of stroke genetics.

To test the protective effects of betaglucin, a novel beta-glucan, on models of myocardial infarction (MI) in rats and dogs.The left anterior descending (LAD) coronary artery occlusion model was used to induce an MI in rats and dogs. Three doses of betaglucin (10, 30 and 100 mg/kg), propranolol (positive control, 1 mg/kg) and vehicle alone (5% glucose solution) were administered before LAD occlusion, and characteristics of the resulting MI were subsequently assessed. In anesthetized dogs, blood pressure, heart rate, ventricular function, coronary artery blood flow and myocardial oxygen consumption were determined before and after the drug administration.The MI mass in both rats and dogs was significantly reduced by betaglucin (30 and 100 mg/kg, P0.01) and propranolol (P0.01). In anesthetized dogs, coronary artery blood flow was increased significantly by betaglucin (30 and 100 mg/kg, P0.01), but blood pressure, heart rate and ventricular function were not changed (P0.05). High-dose betaglucin (100 mg/kg) increased myocardial oxygen consumption, but not to a statistically significant level (P0.05). The hemodynamic indexes were significantly changed by propranolol.Betaglucin has protective effects on myocardial tissue during MI in rats and dogs and has no influence on hemodynamic parameters at a therapeutic dose. The increase in coronary artery blood flow induced by betaglucin might be beneficial in the treatment of patients with MI.

To investigate the effects of allisartan, a new angiotensin II type 1 (AT1) receptor antagonist, on blood pressure (BP) and end-organ damage (EOD) in hypertensive rats and dogs. First, a single dose of allisartan was given intragastrically to evaluate the BP reduction in spontaneously hypertensive rats (SHRs), two kidney-one clip (2K1C) renovascular hypertensive rats and dogs, and Beagle dogs with angiotensin II-induced hypertension. Second, allisartan was mixed in rat chow for long-term treatment. After 4 months of drug administration, rats were instrumented to determine BP and baroreflex sensitivity (BRS). Observation of morphologic changes was used to estimate EOD. Third, the acute toxicity of allisartan was compared with that of losartan in mice. BP was significantly decreased after intragastric administration of allisartan in SHRs, 2K1C rats, 2K1C dogs and Beagle dogs with angiotensin II-induced hypertension. Compared with the control, SHRs that received long-term treatment with allisartan exhibited an improved BRS and organ protective effects. Mice who were administered allisartan experienced less acute toxicity than those treated with losartan. Allisartan is highly effective for BP reduction and organ protection with low toxicity.

To investigate whether the circadian expression of central clock genes and angiotensin II type 1 (AT1) receptors was altered in sinoaortic-denervated (SAD) rats.Male Sprague-Dawley rats underwent sinoaortic denervation or a sham operation at the age of 12 weeks. Four weeks after the operation, blood pressure and heart period were measured in the conscious state in a group of sham-operated (n=10) and SAD rats (n=9). Rest SAD and sham-operated rats were divided into 6 groups (n=6 in each group). The suprachiasmatic nuclei (SCN) tissues were taken every 4 h throughout the day from each group for the determination of the mRNA expression of clock genes (Per2 and Bmal1) and the AT1 receptor by RT-PCR; the protein expression of Per2 and Bmal1 was determined by Western blotting.Blood pressure levels in the SAD rats were similar to those of the sham-operated rats. However, blood pressure variabilities significantly increased in the SAD rats compared with the sham-operated rats. The circadian variation of clock genes in the SCN of the sham-operated rats was characterized by a marked increase in the mRNA and protein expression during dark periods. Per2 and Bmal1 mRNA levels were significantly lower in the SAD rats, especially during dark periods. Western blot analysis confirmed an attenuation of the circadian rhythm of the 2 clock proteins in the SCN of the SAD rats. AT1 receptor mRNA expressions in the SCN were abnormally upregulated in the light phase, changed to a 12-h cycle in the SAD rats.The circadian variation of the 2 central clock genes was attenuated in the SAD rats. Arterial baroreflex dysfunction also induced a disturbance in the expression of AT1 receptors in the SCN.

Aim: To investigate the possible synergism of hydrochlorothiazide and nitrendipine on reducing both blood pressure (BP) and blood pressure variability (BPV) in spontaneously hypertensive rats (SHR). Methods: Seventy animals were randomly divided into seven groups. The doses were 5 and 10 mg/kg for nitrendipine, 10 and 20 mg/kg for hydrochlorothiazide and 10+5, 20+10 mg/kg, respectively, for the combination of these two drugs and 0.8% carboxymethylcellulose as control. The drugs were given via a catheter of gastric fistula. BP was then continuously recorded for 5 h from 1 h before drug administration to the end of 4th hour after drug administration, in conscious and freely moving rats. Results: The effects on both BP and BPV reduction of the combination of hydrochlorothiazide and nitrendipine were greater than the single drug in SHR. The two drugs possessed an obvious synergism on both systolic blood pressure (q=1.79 with small dose and q=1.23 with large dose) and systolic blood pressure variability reduction (q=1.79 with small dose and q=1.39 with large dose) in SHR. Conclusion: The present work clearly demonstrated that there was a synergistic effect between hydrochlorothiazide and nitrendipine in lowering and stabilizing BP in SHR.

Aim: The present work was designed to investigate the effects of nine commonly used antihypertensive drugs on blood pressure (BP) and blood pressure variability (BPV) in conscious sinoaortic-denervated (SAD) rats. Methods: Seventy-two SAD rats were randomly divided into nine groups. They were respectively given nifedipine 3 mg/kg, nitrendipine 5 mg/kg, amlodipine 1 mg/kg, clonidine 10 μg/kg, prazosin 0.5 mg/kg, atenolol 20 mg/kg, telmisartan 20 mg/kg, hydrochlorothiazide 40 mg/kg or captopril 50 mg/kg. The drugs were given via a catheter previously implanted into the stomach. BP was recorded for 5 h from 1 h before drug administration to 4 h after drug administration in conscious, freely moving rats. Results: It was found that all these nine drugs significantly decreased BP in SAD rats. Six of these drugs (nifedipine, nitrendipine, amlodipine, clonidine, prazosin and atenolol) significantly decreased BPV in SAD rats, but the remaining three drugs did not. Clonidine and atenolol increased the heart period and the others did not. No drugs affected the heart period variability. Conclusion: Among nine antihypertensive drugs from different classes, calcium antagonists and sympathetic inhibitors decreased BPV in SAD rats.

Synergistic effects of atenolol and amlodipine for lowering and stabilizing blood pressure in 2K1C renovascular hypertensive rats

Blood pressure, baroreflex sensitivity, and end organ damage in hybrid offspring of spontaneously hypertensive rats and Sprague-Dawley rats

Aim: To demonstrate and compare hemodynamic phenotypes of blood pressure (BP), blood pressure variability (BPV) and baroreflex sensitivity (BRS) in genetic hypertensive rats. Methods: BP was recorded continuously in conscious, freely moving rats using a computerized technique. BPV was expressed as the standard deviation of beat-to-beat BP values during a 1-h period. BRS was determined by measuring the heart period prolongation in response to the elevation in BP produced by an intravenous injection of phenylephrine. Results: Body weight and heart period were not different between spontaneously hypertensive rats (SHR) and stroke-prone spontaneously hypertensive rats (SHR-SP) at the age of 15 weeks. The BP level was markedly higher in SHR-SP than SHR, whereas there were no significant differences in BPV and BRS. Quantitatively, systolic, diastolic and mean BP were significantly elevated by 36.9%, 42.9% and 39.5%, respectively, in SHR-SP compared with SHR ( P P > 0.05). Conclusion: BPV and BRS were not changed in parallel with the BP alterations in SHR and SHR-SP.

Baroreflex dysfunction is associated with a higher rate of sudden death after myocardial infarction (MI). Ketanserin enhances baroreflex function in rats. The present work was designed to examine whether ketanserin improves the post-MI cardiac function and to explore the possible mechanism involved. Spontaneously hypertensive rats (SHR) were treated with ketanserin (0.3 mg·kg−1·d−1). Two weeks later, blood pressure and baroreflex function were measured, followed by a ligation of the left coronary artery. The expressions of vesicular acetylcholine transporter (VAChT) and α7 nicotinic acetylcholine receptor (α7-nAChR) in ischemic myocardium, angiogenesis, cardiac function, and left ventricular (LV) remodeling were evaluated subsequently. Ketanserin significantly improved baroreflex sensitivity (0.62±0.21 vs 0.34±0.12 ms/mmHg, P

To reinvestigate the characteristics of reserpine-induced gastric mucosal lesions (GMLs). The GML-inducing effect of reserpine and the time-course of recovery from reserpine-induced GMLs were examined in Sprague-Dawley (SD) rats. The GML-inducing and blood pressure-decreasing effects of Compound Hypotensive Tablets (CHTs) were investigated in spontaneously hypertensive rats (SHRs). Intracerebroventricular (icv) injection and vagotomy were performed to verify the central vagal mechanism in reserpine-induced GMLs. Single intraperitoneal (ip) injections of reserpine (0.25, 0.5, 1, 2, 4, and 6 mg/kg) dose-dependently induced GMLs in SD rats. Both single and repeated (2 weeks) oral administrations of reserpine led to slight GMLs at doses of 24 mg/kg and 10 mg/kg, respectively. Blood pressure was significantly decreased in SHRs after 2 months of CHT administration (0.01 and 0.03 mg/kg; doses were expressed as the amount of reserpine in the CHT). CHT doses of 0.3 mg/kg induced GMLs, but 0.1 mg/kg did not. Examining the time course of recovery from GMLs, severe GMLs occurred 18 h after ip reserpine (4 mg/kg), obviously lessened at 1 week and healed spontaneously at 3 weeks. Intracerebroventricular injections of reserpine caused GMLs at much lower doses (0.08 and 0.4 mg/kg), and reserpine-induced GMLs were greatly inhibited by vagotomy, suggesting the involvement of a central vagal mechanism. Reserpine-induced GMLs were dose-dependent, and the lesions healed spontaneously within 3 weeks. Long-term treatment with CHT at doses adequate to decrease blood pressure will not induce GMLs. A central vagal mechanism was involved in reserpine-induced GMLs.

Aim: To test the effects of atenolol and amlodipine, either alone or in combination, on blood pressure, blood pressure variability (BPV), baroreflex sensitivity (BRS), and the prevalence of stroke in stroke-prone spontaneously hypertensive rats (SHR-SP). Methods: In the first set of the study, 24 8-month-old, female SHR-SP rats were randomly divided into 3 groups. Blood pressure, heart period, and BRS were determined before and after the intragastric administration of atenolol (10 mg/kg) and amlodipine (1.0 mg/kg), either alone or in combination. In the second set of the study, 40 male and 40 female rats were randomly assigned to 1 of the following 4 groups: control, atenolol (10 mg·kg -1 ·d -1 ), amlodipine (1.0 mg·kg -1 ·d -1 ), and both (10 male and 10 female in each group). The stroke incident and survival time were recorded. Results: Atenolol and amlodipine, either alone or in combination, significantly decreased blood pressure, with the exception of the amlodipine-induced effect on diastolic blood pressure. Meanwhile, only the combination treatment significantly decreased the BPV levels for the same period. The q-values calculated by the probability sum analysis were 1.17 and 2.67 for systolic and diastolic blood pressure, respectively, and were 2.48 and 2.10 for systolic and diastolic BPV, respectively, following administration. Neither drug exhibited any significant effect on BRS. Atenolol and amlodipine, either alone or in combination, significantly increased the lifespan of SHR-SP, with the best effect elicited by the combination therapy. Conclusion: A significant synergism exists between atenolol and amlodipine in lowering and stabilizing blood pressure in SHR-SP. Combination therapy may be an optimal strategy for the prevention of stroke in hypertension.

Aim: To investigate the effect of clonidine, moxonidine, folic acid, and mecobalamin on arterial baroreflex (ABR) function in stroke-prone spontaneously hypertensive rats (SHR-SP) and the possible mechanisms involved. Methods: Eighty-one SHR-SP were divided into 7 groups. Four groups were designated for the intragastric (ig) administration of clonidine (1.0 and 10.0 mug/kg), moxonidine (0.1 and 1.0 mg/kg), folic acid (1.0 mg/kg), and mecobalamin (1.0 mg/kg). Three groups were for the intracerebroventricular (icv) injection of clonidine (4 mug/4 muL), moxonidine (5 mug/4 muL), and mecobalamin (20 mug/4 muL). Blood pressure (BP) was recorded in the conscious state for 30 min and baroreflex sensitivity (BRS) was determined respectively before and after drug administration. Results: Clonidine and moxonidine significantly decreased BP, prolonged the heart period (HP), and increased BRS when administered as either ig or icv injections. Both BP and HP were unchanged by ig folic acid or mecobalamin injection. However, BRS was significantly increased by both. Conclusion: Clonidine, moxonidine, folic acid, and mecobalamin improved impaired ABR function in SHR-SP. The central mechanism was involved in this effect of either clonidine or moxonidine. Mecobalamin improved ABR function through the peripheral mechanism.

To compare high K(+ )-induced contraction and optimal resting tension measured by two commonly used techniques of hypertonic and isotonic K(+ ) in aortas with and without adventitial fat from various age rats.Three age groups of rats (15, 25, and 62 weeks) were used to prepare thoracic aortic rings in which adventitial fat was either removed or left intact. High K(+ ) (30 mmol/L)-induced contractions were observed under increasing resting tensions of 1.0, 1.5, 2.0, 2.5, 3.0, and 3.5 g. Optimal resting tension was the resting tension at which the aorta showed a maximal contraction.The contractions induced by 2 kinds of high K(+ ) were significantly different. Hypertonic and isotonic K(+ ) induced a different style of contraction, and the pattern varied with different ages. At the age of 15 weeks, isotonic K(+ )-induced contractions were greater than hypertonic K+-induced contractions. However, at the age of 62 weeks, isotonic K(+ )-induced contractions were smaller than hypertonic K(+ )-induced contractions. Optimal resting tensions measured by 2 kinds of high K(+ ) were inconsistent. Optimal resting tensions in different kinds of aortic preparations from various age rats were almost a constant of 2 g, determined by isotonic K(+ ), but a variable, determined by hypertonic K(+ ). The adventitial fat could delay the development of high K(+ )-induced contractions at different resting tensions, but had little effect on the maximal contractions.Hypertonic and isotonic K(+ ) may produce different contractions resulting in differences in optimal resting tension in rat aorta.

Aim: The present work was designed to investigate the relationship between hemodynamic parameters and the susceptibility of ventricular arrhythmia to aconitine in conscious Lyon hypertensive rats (LH). Methods: Male LH and Lyon low blood pressure rats (LL) were used. After the determination of baroreflex sensitivity (BRS), ventricular arrhythmia was induced by aconitine infusion in conscious rats. Blood pressure (BP) was recorded during the period of infusion. Results: Compared with the LL rats, the LH rats possessed significantly higher BP, blood pressure variability and lower BRS. The threshold of aconitine required for ventricular fibrillation and cardiac arrest in the LH rats were significantly lower than those in the LL rats. It was found that all the hemodynamic parameters studied were not correlated with the threshold of aconitine required for arrhythmia, with the exception of BRS, which was positively related to the threshold of aconitine required for ventricular premature beat. Conclusion: The LH rats possessed greater susceptibility to aconitine-induced ventricular arrhythmias when compared to the LL rats. This greater susceptibility could not be attributed to any one of the hemodynamic parameters alone studied in the LH rats. It is proposed that various hypertension-associated abnormalities, including the abnormal hemodynamics, may co-contribute to this vulnerability to ventricular arrhythmias.

Aim: To investigate the relationship between arterial baroreflex (ABR) function and telomere length in kidney of rats. Methods: Stroke-prone spontaneously hypertensive rats (SHR-SP) and sinoaortic denervated rats (SAD) were used as models with depressed arterial baroreflex. In the first experiments, SHR-SP rats were examined at the age of 24 weeks for both sexes and 40 weeks for female rats. In the second experiments, SAD rats were studied 4 and 35 weeks after SAD operation. Blood pressure was continuously recorded for 4 h in a conscious state. After the determination of baroreflex sensitivity (BRS), the terminal restriction fragment (TRF) of rat kidney was analyzed using Southern blot. Results: The TRF length was found shorter in: a) male SHR-SP compared with age-matched female SHR-SP; b) female SHR-SP 40 weeks of age compared with 24 weeks of age; c) in rats 35 weeks after operation compared with rats 4 weeks post operation in both sham-operated and SAD rats. Conclusion: In SHR-SP, the TRF length did not correlate with BRS. In addition, SAD did not affect TRF length at either 4 or 35 weeks post-surgery. It may be concluded that baroreflex function does not influence the terminal restriction fragment (TRF) length in rats.

To test the synergistic effects of atenolol and amlodipine on lowering blood pressure (BP) and reducing blood pressure variability (BPV) in 2-kidney, one-clip (2K1C) renovascular hypertensive rats.Forty-eight 2K1C renovascular hypertensive rats were randomly divided into 6 groups. They were respectively given 0.8% carboxymethylcellulose sodium (control), atenolol (10.0 mg/kg), amlodipine (1.0 mg/kg), and combined atenolol and amlodipine (low dose: 5.0+0.5 mg/kg; intermediate dose: 10.0+1.0 mg/kg; high dose: 20.0+2.0 mg/kg). The drugs were given via a catheter in a gastric fistula. BP was recorded for 25 h from 1 h before drug administration to 24 h after administration.Compared with BP before medication, all 3 doses of combined atenolol and amlodipine significantly decreased the BP at 24 h after administration, except for the low dose on diastolic BP. Compared with the control group, all 3 doses of combined atenolol and amlodipine significantly reduced the average BP levels for the 24 h period after administration; furthermore, the high and intermediate doses also significantly decreased the BPV levels for the same period. The q values calculated by probability sum analysis for systolic and diastolic BP for the 24 h period after administration were 2.29 and 1.45, respectively, and for systolic and diastolic BPV for the same period they were 1.41 and 1.60, respectively.There is significant synergism between atenolol and amlodipine in lowering and stabilizing BP in 2K1C renovascular hypertensive rats.

To investigate the blood pressure (BP), baroreflex sensitivity (BRS), and organ damage in hybrids of spontaneously hypertensive rats and Sprague-Dawley rats.Spontaneously hypertensive rats and Sprague-Dawley rats were crossbred, and the F1 hybrids were inbred randomly to produce an F2 generation. At the age of 52 weeks, the F1 and F2 hybrids were tested to determine BP and BRS in a conscious state. Histopathological examinations were carried out after BP recording and BRS studies.BP and BRS were not different in F1 and F2 hybrids. BRS was inversely related to systolic BP (SBP) in male, female, or whole populations of hybrids. Quantitatively, BRS values were one-third determined by SBP level (the determinant coefficient was 0.326). The indexes for left ventricular hypertrophy, aortic hypertrophy, and renal damage were all positively related to BP, and negatively related to BRS. In multiple-regression analysis, left ventricular and aortic hypertrophy and glomerulosclerosis score were all most significantly associated with lower BRS and higher systolic BP. The contribution of BRS to left ventricular and aortic hypertrophy and glomerulosclerosis was greater than that of SBP.The present work with hybrid rats demonstrated quantitatively that the BRS value was one-third determined by SBP level. Both BP level and BRS value contributed greatly to the hypertensive organ damage. However, the contribution of BRS to the hypertensive organ damage was greater than that of BP level in these rats.

To demonstrate and compare hemodynamic phenotypes of blood pressure (BP), blood pressure variability (BPV) and baroreflex sensitivity (BRS) in genetic hypertensive rats.BP was recorded continuously in conscious, freely moving rats using a computerized technique. BPV was expressed as the standard deviation of beat-to-beat BP values during a 1-h period. BRS was determined by measuring the heart period prolongation in response to the elevation in BP produced by an intravenous injection of phenylephrine.Body weight and heart period were not different between spontaneously hypertensive rats (SHR) and stroke-prone spontaneously hypertensive rats (SHR-SP) at the age of 15 weeks. The BP level was markedly higher in SHR-SP than SHR, whereas there were no significant differences in BPV and BRS. Quantitatively, systolic, diastolic and mean BP were significantly elevated by 36.9%, 42.9% and 39.5%, respectively, in SHR-SP compared with SHR (P0.01). However, their variabilities were elevated only by 14.0%, 0.4% and 10.1%, respectively, without statistical significance (P0.05).BPV and BRS were not changed in parallel with the BP alterations in SHR and SHR-SP.

In this issue of Acta Pharmacologica Sinica, we are proud to present to our readers and colleagues a series of review articles from several Chinese-American university laboratories in USA [1–6] . These reviews are a collection of the proceedings from the 8th Chinese Cardiovascular Pharmacology Conference held on July 23–26, 2004 in Urumchi, Xinjiang. As the first of its kind at any Chinese Pharmacological Society meeting, a half-day research symposium in English was held on the first day of this conference (an official meeting of the society held every three years for Chinese cardiovascular pharmacologists worldwide). The symposium was co-sponsored by the Cardiovascular Pharmacology Division of the Chinese Pharmacological Society and the Academy of Cardiovascular Research Excellence (ACRE), a non-profit academic organization in USA that includes many of prominent American faculties from mainland China in the field of cardiovascular research.

To examine the effects of urotensin II, a recently discovered endogenous peptide, on coronary flow (CF), cardiac function, and endothelial nitric oxide synthase (eNOS) expression in isolated rat hearts.Heart was isolated and perfused retrogradely via the aorta in Langendorff mode. Rat urotensin II was administered in the perfusion solution. The eNOS content in myocardium was determined by Western blot.Rat urotensin II had no effect on the heart rate, left ventricular systolic pressure, left ventricular end-diastolic pressure, or +/-dp/dt(max). While rat urotensin II dose-dependently increased CF. CF was increased by 11.43 %, 6.67 %, 6.62 %, 6.56 %, 6.36 %, and 5.86 % respectively in a time-dependent manner at 5, 10, 15, 20, 25, and 30 min after injection of rat urotensin II 6.66 x 10(-2) microg. The maximal effect on CF was found at 5 min following urotensin II administration. N(G)-nitro-L-arginine methyl ester (L-NAME) did not prevent the increased CF in response to urotensin II. Rat urotensin II dose-dependently increased the cardiac eNOS protein expression and this effect was not inhibited by L-NAME.Rat urotensin II did not alter cardiac function but increased CF and the amount of myocardial eNOS protein in the isolated rat heart. The increased CF was independent of the involvement of eNOS.

The present study was designed to test a hypothesis that nonfatal ventricular arrhythmia such as ventricular premature beats (VPB) is a contributing factor in the elevation of blood pressure variability (BPV).Blood pressure (BP) and electrocardiogram were continuously recorded. The relation between VPB and BPV was observed under conscious state in chronic myocardial infarction (MI) rats one month after ligation of the left coronary artery, and further verified under anesthetized state in rat model of ventricular arrhythmia produced by acute intravenous infusion of aconitine.MI rats exhibited a big difference in the count and pattern of VPB, and were divided into no VPB, occasional VPB, and frequent VPB groups. Among the three groups, there were no differences in BP, heart period (HP), and MI size. However, BPV was markedly higher in frequent not occasional VPB rats, and HP variability (HPV) was larger in both frequent and occasional VPB rats, when compared with no VPB rats. In the whole population of MI rats, BPV was positively correlated with VPB and HPV, not with BP, HP and MI size. Infusion of aconitine had no effect on BP, HP, BPV, and HPV during the period without VPB. Frequent VPB after several minutes of aconitine infusion induced significant increase in BPV and HPV with no change in BP and HP. BPV was also positively correlated with VPB and HPV, not with BP and HP. Hemodynamics in aconitine-evoked ventricular tachycardia was characterized as lower BP, higher BPV, and higher HPV.High BPV can be caused by frequent not occasional VPB in rats.

Computerized analysis of blood pressure in conscious freely moving rats is a sound technique for physiological and pharmacological studies. The present work, based on this technique, was designed to introduce two useful methods for the evaluation of antihypertensive drugs in conscious spontaneously hypertensive rat (SHR). They were the directly intragastric administration of drugs and modified probability sum test for evaluating the synergism of the combination of two drugs.(1) Directly intragastric administration was used in conscious rats. A catheter was inserted into stomach immediately after arterial catheter insertion. Three days after operation, blood pressure was recorded and drug might be given intragastrically via the gastric catheter. (2) Modified probability sum test was used to evaluate the synergism of two drugs. The formula was: q=P(A+B)/(PA+PB-PAxB). With this method, it was obtained: q=1.32 for the effects of the combination of atenolol and nitrendipine (20 mg/kg+10 mg/kg) on systolic blood pressure; q=1.41 for the effects of the combination of atenolol and amlodipine (10 mg/kg+1 mg/kg) on systolic blood pressure.The two methods introduced by the present work will be important and useful for antihypertensive drug evaluation in conscious freely moving rats.

Angiotensin II and AT1 receptor are the major effector components of renin-angiotensin system (RAS), and also the main growth-stimulating factors in cardiovascular system. The present study was to observe these two factors in the hypertrophied ventricles and aortas of sinoaortic-denervated rats.Rats were examined at 2, 10, and 16 weeks after sinoaortic denervation (SAD). The hypertrophy was evaluated by the ratio of organ weight to body weight. Angiotensin II concentration and AT1 receptor mRNA expression were measured by radioimmunoassay and RT-PCR respectively, using a positive control of candesartan treatment.Aortic hypertrophy existed in 2-, 10-, and 16-week SAD rats, left ventricular hypertrophy in 10- and 16-week SAD rats, and right ventricular hypertrophy in 16-week SAD rats. In all three kinds of examined SAD rats, plasma angiotensin II levels remained unchanged, indicating circulating RAS is at normal level in the chronic phase of SAD. However, cardiovascular tissue RAS was activated, as evidenced by increase of aortic angiotensin II concentrations at 10 and 16 weeks after SAD, and up-regulation of aortic and left ventricular AT1 receptor mRNA expressions at 16 weeks after SAD.The activated tissue RAS is secondary to the hypertrophy, and probably involved in the maintenance of cardiovascular hypertrophy following SAD.

To construct an AAV based vector carrying human endothelial nitric-oxide synthase (eNOS) cDNA and study its expression in vitro for future gene therapy.eNOS cDNA was inserted into the EcoR I site of pSNAV-1 containing the cytomegalovirus (CMV) promoter and inverted terminal repeat sequences of adeno-associated virus. The constructed vector was transfected into BHK and C2C12 cells. eNOS cDNA and mRNA were detected by polymerase chain reaction (PCR) and reverse transcription-PCR (RT-PCR), respectively.By restriction enzyme digestion analysis, it was proved that eNOS cDNA was inserted into pSNAV-1 in a proper direction. PCR detection demonstrated that pSNAV-eNOS was transferred into both BHK and C2C12 cells. RT-PCR analysis showed that these pSNAV-eNOS transfected cells could express eNOS mRNA.pSNAV-eNOS was successfully constructed with the ability to express human eNOS mRNA in cultured mammalian cells.

To study the effect of candesartan cilexetil (candesartan), a new AT1 receptor antagonist, on sinoaortic denervation (SAD)-induced cardiovascular hypertrophy and its potential mechanisms in rats.For long-term treatment, candesartan (6 mg/kg/d) was given in rat food for 16 weeks after SAD surgery, and for acute treatment, a single dose of candesartan (3 mg/kg) was administrated intragastrically at 30 d after SAD.The indexes of left ventricular and aortic hypertrophy in candesartan-treated SAD rats were decreased when compared with untreated SAD rats, and similar to or less than those in normal rats. SAD-induced cardiomyocyte hypertrophy, myocardial fibrosis, wall thickening of intramyocardial arterioles and aortae, and destruction of vascular internal elastin membrane were almost inhibited by candesartan. The plasma angiotensin II levels were markedly increased in treated SAD rats and negatively correlated with the indexes of hypertrophy. Both blood pressure and its variability were reduced by a single dose of candesartan during 3 h of observation period.Candesartan can efficiently inhibit SAD-induced cardiovascular hypertrophy. In addition to known mechanisms, upregulation of circulating angiotensin II and stabilization of blood pressure may be involved in this cardiovascular protection of candesartan.

Arterial baroreflex (ABR) is a very important mechanism in the regulation of cardiovascular activities. As ABR function is largely inhibited by anesthesia, its measurement in conscious animal becomes important. The present review summarizes the works concerning ABR function in conscious rats completed in our department in the last 10 years. Firstly, a new method was established to measure arterial baroreflex-blood pressure control (ABR-BP). ABR-BP and baroreflex sensitivity measured with classic method are two different parts of the ABR function. Secondly, it was proposed that ABR function predicted the end-organ damage in hypertension. Thirdly, interruption of ABR induced severe end-organ damages. Increased blood pressure variability (BPV) and activation of renin angiotensin system were involved in the mechanisms underlying organ damages in sinoaortic denervation (SAD) rats. Fourthly, we propose that amelioration of ABR function may serve as a new strategy for improving the prognosis of cardiovascular diseases. Ketanserin improved the impaired ABR function in SHR. Finally, the possibility to develop a strain of rats with spontaneous deficiency on ABR function is mentioned.

To study the determination of arterial baroreflex-blood pressure control (ABR-BP) in conscious rats.Blood pressure was continuously recorded with a computerized system in conscious freely moving rats. The principle of ABR-BP measurement is to compare the pressor response to a vasoactive drug (angiotensin II) before and after the interruption of this reflex.(1) ABR-BP values revealed by angiotensin II were closely correlated with those by phenylephrine. Doses of angiotensin II did not influence the results within certain range. (2) ABR-BP was well correlated with arterial baroreflex-heart period control (ABR-HP). (3) Anesthesia inhibited ABR-BP. There existed a circadian variation of ABR-BP in WKY rats. (4) Blood pressure variability was closely related to ABR-BP, but not to ABR-HP. (5) ABR-BP was impaired in hypertensive rats.ABR-BP is an important parameter to reflect the function of ABR. The present work makes it possible to determine ABR-BP in conscious rats. ABR-BP plays an important role in maintaining blood pressure stability and it is impaired in hypertension.