The aim of this study was to observe the change in angiotensin II receptor subtype 1 (AT(1)) autoantibody during the development of renovascular hypertension (RVH). The Goldblatt renovascular hypertension model was established by the two-kidney one-clip method, and a synthetic peptide corresponding to amino acid sequence 165-191 of the second extracellular loop of the AT(1)-receptor was used as the antigen. Sera AT(1)-receptor autoantibody was detected by SA-ELISA. It was shown that two weeks after operation both the frequency of occurrence and the titre of autoantibodies to AT(1)-receptor were significantly increased as compared with the pre-treatment control. The increase in autoantibodies lasted several weeks and then decreased gradually to the pre-clipping level at 12 weeks. It is suggested that autoimmune mechanisms are involved in the pathogenesis of renovascular hypertension and the AT(1) autoantibodies may be one of the mechanisms leading to cardiac hypertrophy.
Animals ; Autoantibodies ; blood ; Disease Models, Animal ; Hypertension, Renovascular ; blood ; immunology ; Kidney ; physiopathology ; Rats ; Receptor, Angiotensin, Type 1 ; immunology ; metabolism

Experiments were performed in male Sprague-Dawley rats (150-200 g). A silver clip (0.2 mm internal diameter) was placed on the left renal artery of rats. After operation the rats were divided into 4 groups sham group, 2K1C (two-kidney one clip) group, 2K1C+Arg (2K1C and L-Arg 150 mg/kg x d(-1) by drinking) group, and 2K1C+NAME (2K1C and L-NAME 10 mg/kg x d(-1) i.p.) group. The animals were studied at two time points (4 and 7 weeks after operation) corresponding to phases I and II of Goldblatt hypertension. The animals were deeply anesthetized with pentobarbital and perfused by the cardiac route with saline (100 ml) and freshly prepared 4% paraformaldehyde in phosphate buffer (300 ml). The caudal medulla was removed, then placed in 25% sucrose in PB for 12 h in a 4 degrees C fridge. The 40 microm coronal slices of brainstems were cut with cryostat, collected in PBS for nNOS study by immunohistochemistry. The results showed that (1) only a few neuronal nitric oxide synthase (nNOS) positive neurones were found in caudal medulla, including the depressor area of the ventral surface of medulla oblongata (VSMd) and the caudal pressor area (CPA) of the sham operated animals. The number of nNOS positive neurons in caudal medulla was significantly increased in 2K1C Goldblatt hypertension rats at 4 and 7 weeks. (2) The number of nNOS positive neurons in VSMd and CPA were 2K1C+Arg > 2K1C >2K1C +NAME > sham. (3) L-Arg enhanced the expression of nNOS whereas L-NAME inhibited the expression of nNOS in caudal medulla. (4) The character of nNOS expression was similar in Goldblatt hypertension rats at 4 weeks with that of the rats at 7 weeks.
Animals ; Hypertension, Renovascular ; enzymology ; physiopathology ; Male ; Medulla Oblongata ; enzymology ; metabolism ; Nitric Oxide Synthase ; biosynthesis ; Rats ; Rats, Sprague-Dawley

The present study was to investigate the mRNA, protein expression and the activity of calcineurin in the hypertrophic heart, and to determine the effect of calcineurin inhibitor--cyclosporine A (CsA) on the regression of cardiac hypertrophy in renovascular hypertensive rats. Renovascular hypertension was induced by two kidney-one clip methods. Two months after the operation, cardiac hypertrophy was determined by histological analysis performed in some rats (2K1C-2M), then the rats were subdivided into 2 groups: (1) 3-month old two kidney-one clip group (2K1C-3M) with rats receiving 0.9% NaCl per day for one month, and (2) CsA-treated group with rats treated with CsA for one month. Sham-operated rats were used as control. The ratio of the left ventricular weight to tibial length (LVW/TL), the area of cardiac myocyte, mRNA and protein expression and the activity of calcineurin were determined. Both the LVW/TL and the cardiomyocyte area were significantly larger in 2K1C-2M and 2K1C-3M rats than in age-matched sham-operated rats. Treatment with CsA significantly attenuated the increase in the LVW/TL as well as the cardiomyocyte area. The mRNA, protein expression and the activity of calcineurin were significantly higher in 2K1C-2M and 2K1C-3M rats than those in the age-matched sham-operated rats, while the elevation of mRNA, protein expression and activity of calcineurin were significantly suppressed in the CsA-treated rats. In conclusion, calcineurin plays a role in the progression of cardiac hypertrophy in renovascular hypertensive rats. The inhibition of calcineurin can reverse cardiac hypertrophy.
Animals ; Calcineurin ; biosynthesis ; genetics ; metabolism ; Cyclosporine ; pharmacology ; Hypertension, Renovascular ; complications ; metabolism ; physiopathology ; Hypertrophy, Left Ventricular ; etiology ; metabolism ; physiopathology ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Sprague-Dawley

The present study was aimed at examining the regulation of aquaporin (AQP)-2 water channels in the kidney in two-kidney, one clip (2K1C) hypertension. Rats were made 2K1C hypertensive for 6 weeks, and their expression of AQP2 channel proteins was determined in the clipped and contralateral kidneys. To examine the upstream affecting AQP2 channels, adenylyl cyclase activity was also determined. Along with the hypertension, in the clipped kidney, the abundance of AQP2 proteins was significantly decreased in the cortex, outer and inner medulla, while their trafficking remained unaltered. Concomitantly with the reversal of the blood pressure at 24 hours following removal of the clip, the AQP2 abundance also returned to the control level. The arginine vasopressin-evoked generation of cAMP was decreased in the clipped kidney, which again was reversed to the control level following removal of the clip. In contrast, the expression of AQP2 channels as well as the activity of adenylyl cyclase remained unaltered in the contralateral kidney. These results indicate an altered regulation of AQP2 water channels in the clipped kidney in 2K1C hypertension.
Adenylate Cyclase/metabolism ; Animal ; Aquaporins/*analysis ; Blood Pressure ; Cyclic AMP/biosynthesis ; Hypertension, Renovascular/*metabolism ; Kidney/*chemistry ; Male ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo explore the immuno-neurologic regulation of hypertension and its inherent law as well as the mechanisms of curing and systemic regulating effect of ZiShuiJiangHuoYin(ZSJHY).

METHODTo detect expression level of AT-1mRNA in two-kidney-one-clamp renal hypertension rat lymphocyte cell by means of RT-PCR.

RESULTThe level of AT-1 mRNA in lymphocyte was higher in group of 2K1C-RHR than that in normal group. ZSJHY (20 g/kg, 40 g/kg) had regulating effect on this.

CONCLUSIONDepressing excessive expression of lymphocyte AT-1 mRNA may be one of the mechanisms where ZSJHY exert immunoregulation action.

Animals ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Hypertension, Renovascular ; metabolism ; Lymphocytes ; metabolism ; Male ; Plants, Medicinal ; chemistry ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar ; Receptors, Angiotensin ; biosynthesis ; genetics

OBJECTIVETo study the changes in the mRNA expression of endothelial cellular adhesion molecules in the cerebral blood vessels in rats with prestroke condition caused by simulated cold wave.

METHODSTwo-kidney two-clip renovascular hypertension was induced in 48 male SD rats, which were subsequently randomly assigned into cold wave exposure and non-exposed group (n=24). Each group was further divided into 4 sub-groups according to their systolic blood pressure, namely the sham-operated group with blood pressure (BP)<140 mmHg, mild hypertensive group with BP of 160-199 mmHg, moderate hypertensive group with BP of 200-219 mmHg, and severe hypertensive group with BP no less than 220 mmHg. Cold wave exposure was simulated by housing the rats in an artificial climate chamber with 3 cycles of 12 h light at 22 degrees celsius; and 12 h dark at 4 degrees celsius;. The non-exposed group was kept at 22 degrees celsius; throughout the experiment. After the exposure, the rats were sacrificed and the tissues of the frontal lobe were slice into 2.0-mm-thick coronal sections for real-time RT-PCR detection of vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), and p-selectin mRNA expressions. The 5.0-microm-thick frozen sections from the bregma section underwent in situ hybridization of VCAM-1, ICAM-1, and p-selectin. The other sections were stained with HE to observe the infarct lesions, and the rats with cerebral infraction were excluded from the statistical analysis.

RESULTSIn rats with cold wave exposure-induced prestroke condition and BP <220 mmHg, VCAM-1, ICAM-1, and p-selectin mRNA expressions all increased compared with those in the non-exposed group. In rats with BP>or=220 mmHg and cold exposure, the expressions all decreased to some extent compared with those in the non-exposed treatment. In the non-exposed rats, a positive correlation of BP to VCAM-1, ICAM-1, and p-selectin mRNA expressions were noted, and this correlation was also found in cold-wave-exposed rats with BP <220 mmHg; VCAM-1, ICAM-1, and p-selectin mRNA expressions decreased dramatically in the exposed rats with BP >or=220 mmHg compared with those in rats with BP <220 mmHg.

CONCLUSIONPersistent and severe hypertension impairs the modulatory function of the cerebral vascular endothelia, which is a prerequisite for the stroke vulnerability. The modulatory function deteriorates as the BP further increases.

Animals ; Atmosphere Exposure Chambers ; Cerebral Arteries ; metabolism ; Cold Temperature ; Environment, Controlled ; Equipment Design ; Hypertension, Renovascular ; complications ; metabolism ; Intercellular Adhesion Molecule-1 ; genetics ; metabolism ; Male ; P-Selectin ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Stroke ; etiology ; metabolism ; Vascular Cell Adhesion Molecule-1 ; genetics ; metabolism

This experiment on rats was aimed to investigate the expression of intermedin (IMD) in hypertrophic cardiac myoctye of renal vascular hypertension induced by incomplete ligation of the left renal artery, and so to detect and compare the changes of the expression after administration of Valsartan, Amlodipine and Enalapril respectively. The criterion for standard modeling was systolic pressure > or = 140 mmHg. At 4 weeks after successful modeling, 60 SD male rats were randomly divided into 5 groups, namely the hypertrophy group, the 3 drug-treatment groups, and the sham-operation group as control. Blood pressure, left ventricular mass index (LVMI), and the left ventricular mean transverse diameter of myocardial cell (LVTDM) were investigated at the 10th week after model establishment. Gene expression of IMD mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR), and the optical density of the band was measured by use of the Gel Documentation System. The ratio of IMD mRNA to beta-actin mRNA was considered the relative amount of IMD. When compared with control, the blood pressure increased significantly in the hypertrophy group. There was no statistically significant difference between the treatment groups. No significant difference in heart rate was noted at 4 weeks after operation in all groups. LVMI and LVTDM levels were significantly higher in the hypertrophy group than in the other groups; LVMI and LVTDM levels showed no significant difference among the treatment groups but they were obviously higher than those of the Sham-operation group. The gene expression of IMD mRNA in the hypertrophy group was upregulated in the myocardium, when compared with that in the other groups. Meanwhile, although IMD mRNA in the treament groups was higher than that in the Sham-operation group, no statistically significant difference of myocardial IMD mRNA was found between the treament groups. These results suggested that, in this experiment, intracardiac IMD mRNA was upregulated and could participate in the regulation of cardiac remodeling in renal vascular hypertension-induced cardiac hypertrophy. This upregulation could improve the pathologic and physiologic process of cardiac hypertrophy, and could associate with the pressure loading or myocardia hypertrophy. However, the change did not display any difference that could be attributed to the variety of hypotensive drugs.
Adrenomedullin ; genetics ; metabolism ; Amlodipine ; therapeutic use ; Animals ; Antihypertensive Agents ; therapeutic use ; Cardiomegaly ; etiology ; metabolism ; Enalapril ; therapeutic use ; Hypertension, Renovascular ; complications ; drug therapy ; metabolism ; Male ; Myocardium ; metabolism ; Neuropeptides ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Tetrazoles ; therapeutic use ; Valine ; analogs & derivatives ; therapeutic use ; Valsartan

To investigate the effect of losartan on the axis of prolylcarboxypeptidase (PRCP)--kallikrein of the two-kidney, one-clipped (2K1C) hypertensives rats, and explore the novel protection mechanism of losartan on the kidney. Sprague-Dawley (SD) rats were used to develop the 2K1C hypertensive rats. Then, the rats were treated with prazosin (5 mg x kg(-1) x d(-1)) or losartan (5, 15 and 45 mg x kg(-1) x d(-1)) or vehicle, separately. At the same time, the blood pressures were observed. After treated for four weeks, the ratio of right kidney weight and body weight, the change of glomerular morphology, and K+, Na+, creatinine and blood urea nitrogen (BUN) of the serum were used for evaluation of kidney. The expressions of PRCP mRNA in the kidneys were determined by RT-PCR. The protein levels of PRCP, tissue kallikrein, plasma kallikrein, TGF-beta1 in kidney or plasma were measured by Western blotting. Results showed that the changes of body weight and kidney weight ratio, glomerular fibrosis degree and the biochemistrical index of serum induced by hypertension were relieved when the hypertensive rats treated with losartan for four weeks. Meanwhile, treatment of losartan also significantly decreased expression of TGF-beta1 and increased expressions of PRCP, plasma kallikrein and tissue kallikrein. The protective effects of losartan on the kidney of 2K1C hypertensive rats are activation of the axis of PRCP-kallikrein and reducing the expression of TGF-beta1.
Animals ; Antihypertensive Agents ; pharmacology ; Blood Pressure ; drug effects ; Carboxypeptidases ; genetics ; metabolism ; Hypertension, Renovascular ; metabolism ; pathology ; physiopathology ; Kallikreins ; blood ; metabolism ; Kidney ; metabolism ; pathology ; Kidney Glomerulus ; pathology ; Losartan ; pharmacology ; Male ; Organ Size ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta1 ; blood ; metabolism

AIMTo investigate the preventive and reversional effect of praeruptorum caumarin compound on left ventricular hypertrophy in renovascular hypertensive rats (RHR) and its mechanism.

METHODSThe two-kidney-one-clip (2K1C) RHR model was used. The blood pressure, wet weight of the left ventricle, surface area of myocardial cells, resting [Ca2+]i level and Na+, K(+)-ATPase, Ca2+, Mg(2+)-ATPase activity of myocardial membrane and mitochondria were measured.

RESULTSPraeruptorum caumarin 30 mg.kg-1.d-1 was given ig for 9 weeks from the 6th or 9th week after operation in the preventive or regressive group. The blood pressure, left ventricle wet weight and area of myocardial cells of the preventive and regressive group were significantly reduced than that of the LVH group. The resting [Ca2+]i of the both praeruptorum caumarin treated groups (121 +/- 13, 133 +/- 9 nmol.L-1) were lower than that of the LVH group (158 +/- 7 nmol.L-1). The KCl-induced [Ca2+]i elevation was decreased more significantly in preventive and regressive group than that of the hypertrophic myocytes. The activity of Na+, K(+)-ATPase and Ca2+, Mg(2+)-ATPase increased by 40% and 93% in the preventive group, 28.4% and 48.8% in regressive group than that of the LVH group.

CONCLUSIONPraeruptorum caumarin was shown to prevent and reverse hypertrophy of LVH by lowering [Ca2+]i and increasing the ATPase activity.

Animals ; Apiaceae ; chemistry ; Ca(2+) Mg(2+)-ATPase ; metabolism ; Calcium ; metabolism ; Cell Separation ; Coumarins ; isolation & purification ; pharmacology ; therapeutic use ; Disease Models, Animal ; Hypertension, Renovascular ; complications ; metabolism ; pathology ; prevention & control ; Hypertrophy, Left Ventricular ; etiology ; metabolism ; pathology ; Mitochondria ; enzymology ; Myocytes, Cardiac ; drug effects ; metabolism ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley ; Sodium-Potassium-Exchanging ATPase ; metabolism

The purpose of this study is to evaluate the effects and the underline mechanisms of berberine on the cardiac function and left ventricular remodeling in rats with renovascular hypertension. The renovascular hypertensive model was established by the two-kidney, two-clip (2K2C) method in Sprague-Dawley (SD) rats. Two weeks after surgery, all the operated SD rats were randomly assigned into four groups: (1) renovascular hypertensive model group; (2) berberine 5 mg x kg(-1) group; (3) berberine 10 mg x kg(-1) group; (4) captopril 45 mg x kg(-1) group; and the sham operated rats were used as control. Four weeks after the drugs were administered, the cardiac function was assessed. The ratios of heart weight to body weight (HW/BW), left ventricular weight to body weight (LVW/BW) and right ventricular weight to body weight (RVW/BW) were compared between groups. Coronal sections of the left ventricular tissue (LV) were prepared for paraffin sections, picrosirius red and HE staining was performed. The left ventricular wall thickness (LVWT), interventricular septal thickness (IVST), the parameters of myocardial fibrosis indicated by interstitial collagen volume fraction (ICVF) and perivascular collagen area (PVCA) were assessed. Nitric oxide (NO), adenosine cyclophosphate (cAMP) and guanosine cyclophosphate (cGMP) concentrations of left ventricular tissue were measured. Berberine 5 mg x kg(-1) and 10 mg x kg(-1) increased the left ventricular +/- dp/dt(max) and HR. Berberine 10 mg x kg(-1) decreased HW/BW and LVW/BW. The image analysis showed that both 5 and 10 mg x kg(-1) of berberine decreased LVWT, ICVF and PVCA, while increased the NO and cAMP contents in left ventricular tissue. Berberine could improve cardiac contractility of 2K2C model rats, and inhibit left ventricular remodeling especially myocardial fibrosis in renovascular hypertension rats. And such effects may partially associate with the increased NO and cAMP content in left ventricular tissue.
Animals ; Berberine ; pharmacology ; Collagen ; metabolism ; Cyclic AMP ; metabolism ; Cyclic GMP ; metabolism ; Hypertension, Renovascular ; metabolism ; physiopathology ; Male ; Myocardium ; metabolism ; pathology ; Nitric Oxide ; metabolism ; Organ Size ; drug effects ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Ventricular Function, Left ; drug effects ; Ventricular Remodeling ; drug effects