Production of lysophosphatidic acid by lysophospholipase D in incubated plasma of spontaneously hypertensive rats and Wistar Kyoto rats.

Lysophosphatidic acid has been identified as a vasopressor principle in incubated mammalian plasma and sera, and shown to be generated extracellulary by lysophospholipase D-like activity. In this study, we monitored the time course of changes in the major phospholipid fractions during incubation of plasma, and found that polyunsaturated lysophosphatidic acids accumulate more rapidly than saturated lysophosphatidic acids at expense of the corresponding lysophosphatidylcholines. We compared the phospholipase activities for producing bioactive LPA in age-matched spontaneously hypertensive rats and Wistar Kyoto rats. The lysophospholipase D activity in rat plasma was found to be independent of strain and age. We suggest that lysophospholipase D functions in rat for persistent production of bioactive LPA in the circulation throughout life. However, our finding that production of LPA in spontaneously hypertensive rats was not greater than that in Wistar Kyoto rats does not seem to support the idea that increased production of LPA is involved in the pathogenesis of hypertension.     

Spontaneously hypertensive and Wistar Kyoto rats are genetically disparate.

Spontaneously hypertensive rats (SHR) are one of the most common animal models used to study essential hypertension in humans. Because SHR and normotensive Wistar Kyoto (WKY) rats were both established from the same parental, normotensive Wistar stock, WKY animals have been used almost exclusively as control animals in studies of SHR. Recently, the suitability of WKY rats as normotensive controls for SHR has been challenged. To establish whether or not SHR and WKY rats share the same immunologic backgrounds, we initially performed a series of skin grafting experiments on these animals. In all cases, grafts of SHR donor skin to WKY recipients and of WKY donor skin to SHR recipients resulted in complete rejection within 7 to 10 days. In addition, grafts of WKY donor skin to other WKY recipients resulted in graft rejection. By contrast, skin grafts between SHRs were always accepted. To further characterize the genetic distinctions between SHR and WKY rats, allelic profiles based on a series of immunologic and biochemical markers were established for each strain. These findings clearly establish that SHR and WKY rats differ at the major histocompatibility complex, in specific blood group antigens, and in a panel of isozymic markers. Moreover, whereas SHRs have the same genetic profiles irrespective of source, some colonies of WKY rats are outbred, as judged by their variant allelic profiles.     

Protein kinase C activity in platelets from spontaneously hypertensive rats (SHR) and normotensive Wistar Kyoto rats (WKY)

Calcium-activated phospholipid dependent protein kinase (protein kinase C) activity in platelets was measured in 4, 12, and 20-week-old SHR and WKY. At age 4-weeks, there was no significant difference in protein kinase C activity and systolic blood pressure between SHR and WKY. In 12 and 20-week-old SHR, both protein kinase C activity and systolic blood pressure were significantly higher than in the age-matched WKY. These results suggest that protein kinase C may be involved in the control of blood pressure in SHR and WKY.     

Kidney adaptation in nitric oxide-deficient Wistar and spontaneously hypertensive rats

We investigated the renal structural and functional consequences of nitric oxide (NO) deficiency co-treated with angiotensin-converting enzyme inhibitor (ACEi) in 20 adult male Wistar rats and 20 spontaneously hypertensive rats (SHR). The animals were separated into eight groups (n = 5) and treated for 30 days: Control, L-NAME (NO deficient group), Enalapril, L-NAME + Enalapril. The elevated blood pressure in NO deficient rats was partially reduced by enalapril. Serum creatinine was elevated in L-NAME-SHRs and effectively treated with enalapril. The proteinuria was significantly higher only in L-NAME-SHRs, and this was reduced by treatment with ACEi. The glomerular volume density (Vv(gl)) in L-NAME rats, both Wistar and SHR, was greater than in matched control rats, and enalapril treatment effectively prevented this Vv(gl) increase. No significant differences were observed in tubular volume density, Vv(tub), or tubular surface density, Sv(tub), in all Wistar groups. The Vv(tub) was smaller in L-NAME-SHRs than in control SHRs, and this tubular alteration was not prevented by enalapril. The Sv(tub) was not different among the SHR groups. In Wistar rats no changes were seen in vascular surface density, but a greatly increased cortical vascular volume density was seen in the enalapril treated rats. The vascular length density was greatly diminished in NO deficient rats that was effectively prevented with enalapril treatment. The vascular cortical renal stereological indices are normally reduced in SHRs. Administration of enalapril, but not L-NAME, changed this tendency. However, enalapril was not totally effective in preventing vascular damage in SHR NO deficient animals.     

Effects of nitric oxide synthase inhibitor on tyrosine hydroxylase mRNA in the adrenal medulla of spontaneously hypertensive and Wistar Kyoto rats.

We studied the effects of N(G)-nitro-l-arginine methyl ester (L-NAME) on catecholamine levels, tyrosine hydroxylase (TH) activity, and TH mRNA levels in the adrenal medulla of spontaneously hypertensive rats (SHR) and Wistar Kyoto rats (WKY). L-NAME (100 mg/L in drinking water) and atropine (10 mg/L in drinking water) were administered for 2 weeks. Epinephrine and norepinephrine levels, TH activity, and TH mRNA levels in the adrenal medulla of L-NAME-treated WKY were significantly decreased. These parameters were not significantly altered in the adrenal medulla of L-NAME-treated SHR. Nitrite/nitrate levels in the adrenal medulla of L-NAME-treated WKY were significantly decreased; however, no significant change in L-NAME-treated SHR was observed. Ca(2+)-dependent nitric oxide synthase (NOS) activity in the adrenal medulla of SHR was significantly decreased compared to that of WKY. TH mRNA levels in L-NAME + atropine-treated and L-NAME-treated WKY were significantly lower than TH mRNA levels in control WKY. These results suggest that nitric oxide in the adrenal medulla may enhance the catecholamine biosynthetic pathway via increased TH mRNA expression. Our results also suggest that this effect is suppressed in SHR due to decreased NOS activity in the adrenal medulla.     

Protein kinase C activity and reactivity to phorbol ester in vascular smooth muscle from spontaneously hypertensive rats (SHR) and normotensive Wistar Kyoto rats (WKY)

Protein kinase C (PKC) activity in aortic and renal arterial smooth muscle from SHR (20-23 wk male; mean arterial pressure = 178 mm Hg) and WKY (age/sex matched; mean arterial pressure = 126 mm Hg) was quantitated. Activity was greatest in the particulate fractions relative to the soluble fractions in all sources. The only difference between SHR and WKY was in the soluble fraction from SHR renal arteries, which had 2 fold more activity (255 pmol/mg/min) when compared with WKY (136 pmol/mg/min). This difference was not apparently related to force modulation, since the magnitude of isometric force development in renal arteries in response to phorbol 12,13-dibutyrate was not different between SHR and WKY. The magnitude of force developed in response to phorbol 12,13-dibutyrate and PKC activity in the particulate fraction was greatest in aorta vs. renal arteries in both WKY and SHR. These results suggest that regional vascular differences in the amount of PKC activity may exist which are not apparently related to a disease state (i.e., hypertension). These differences may be related to differential sensitivity to phorbol ester-mediated contractions in isolated smooth muscle.     

Fatty acid composition of brain hemispheres of spontaneously hypertensive rats suckled by female Wistar rats

Total lipid fatty acid composition was investigated in brain hemispheres of male Spontaneously Hypertensive Rats (SHR), compared with normotensive Wistar Kyoto rats (WKY) used as controls. Both strains were suckled by adoptive Wistar mothers, and then fed a standard diet after weaning. No difference was observed between the two hemispheres of WKY killed either at 10 or 30 days. In SHR killed at 10 days, the two hemispheres showed differences, SHR left hemispheres exhibiting greater fatty acid composition changes than those of WKY, phenomenon that toned down at 30 days. Hence, SHR pups showed a different total lipid fatty acid composition of their brain hemispheres when compared with their WKY controls, though the two strains received the same diet. Genetically programmed hypertension might be, directly or not, involved in these changes.     

Effects of endothelin 3 on diastolic blood pressure of pithed Sprague-Dawley, Wistar Kyoto, and spontaneously hypertensive rats before and after pretreatment with nifedipine

Pressor actions of endothelin 3 (ET3) were examined in pithed Sprague-Dawley (SD), Wistar-Kyoto (WKY), and spontaneously hypertensive (SH) rats before and after the administration of the calcium channel antagonist, nifedipine. Systolic and diastolic blood pressures were recorded via an intra-arterial catheter from sodium pentobarbital anaesthized rats prior to pithing. The systolic and diastolic blood pressures recorded from SH rats were significantly greater than those of SD and WKY rats; however, after pithing there were no significant differences between the diastolic blood pressures among the various strains. Administration of nifedipine significantly reduced the diastolic blood pressure of pithed rats to an equal extent in all three strains. The infusion of ET3 produced a dose-dependent increase in diastolic blood pressure of SD, WKY, and SH rats, but neither vascular sensitivity nor reactivity to ET3 was altered in SH rats. Nifedipine was more effective at inhibiting the vasoactive actions of ET3 in SD and WKY than in SH rats. It was therefore concluded that the pressor actions of ET3 in SH rats may be less dependent on the influx of calcium through a dihydropyridine-sensitive calcium channel as compared with WKY and SD rats.     

Stress ulcer and open-field behavior of spontaneously hypertensive, normotensive, and Wistar rats

Spontaneously hypertensive rats (SHRs) and the normotensive Wistar/Kyoto (WKY) rat were observed, along with Wistar rats (which represent the parent strain), on various open-field behaviors. All three strains were subsequently exposed to the activity-stress (A-S) ulcerogenic procedure. SHR and Wistar rats were very active in most open-field measures as compared with WKY rats, but only SHRs were active during the A-S treatment. WKY rats were very ulcer prone and had significantly more ulcers than SHRs, which in turn had more ulcers than Wistar rats. It was anticipated that Wistar rats would resemble the WKY rats, but in most measures the Wistars resembled the SHRs. The study suggests that although WKY rats function as an appropriate control for hypertension studies, these rats may be inappropriate as controls for other physiological and behavioral studies.     

Dietary fat source and cholesterol interactions alter plasma lipids and tissue susceptibility to oxidation in spontaneously hypertensive (SHR) and normotensive Wistar Kyoto (WKY) rats

Due to the potential for dietary fat source to alter plasma lipids and tissue antioxidant status, we hypothesized that blends of saturated, n-6 and n-3 fats with cholesterol would affect LDL and tissue susceptibility to in vitro oxidation. The effects of dietary fat blends of butter (B), beef tallow (T), soybean oil (SBO) or menhaden oil (MO) and cholesterol on systolic blood pressure (SBP), plasma lipoproteins and tissue susceptibility to glutathione (GSH) depletion and lipid peroxidation (TBARS) were examined in spontaneously hypertensive (SHR) and Wistar Kyoto (WKY) rats. SBP in SHRs was higher (p < 0.001) than in WKYs at 13-weeks of age but was not altered by dietary fat or cholesterol. LDL- and HDL-cholesterol were greater (p < 0.001) in WKY than SHR. LDL-cholesterol and (VLDL7- + LDL-cholesterol)/HDL-cholesterol ratios were reduced in MO vs. B, T and SBO groups. HDL-cholesterol levels tended to be lower and greater in B and MO groups, respectively vs. T and SBO groups. Initial LDL fluorescence was greater (p < 0.001) in high- vs. low-cholesterol groups. The change in LDL fluorescence was reduced (p < 0.001) in high-cholesterol groups, and MO vs. B, T and SBO rats. MO fed rats had reduced (p < 0.001) RBC, heart and liver GSH depletion and reduced (p < 0.01) tissue TBARS and RBC MDA production. In summary, a moderate level of dietary MO did not increase tissue and LDL in vitro oxidizability in SHR and WKY rats. High dietary cholesterol exhibited a protective effect against in vitro oxidation of LDL and selected tissues.     

Differential size distribution of atrial dense granules in spontaneously hypertensive, Wistar-Kyoto and Wistar rats

The differential size distribution of atrial dense granules (ADGs) in spontaneously hypertensive rats (SHR) and two normotensive controls, Wistar and Wistar-Kyoto (WKY) rats, was investigated. The ADGs in SHR were smaller than those in Wistar rats. The ADGs of WKY rats were of intermediate size. It is possible that ADGs are more rapidly secreted in SHR compared with WKY and Wistar rats or that the smaller-diameter granules may contain more atriopeptinogen than the larger granules. The intermediate size of the ADGs in WKY suggests that the WKY variant is morphologically intermediate between the SHR and Wistar strains.     

自发性高血压大鼠和Wistar大鼠脑动脉平滑肌细胞膜电流的比较

目的:探讨自发性高血压大鼠(SHR)和Wistar大鼠脑动脉(BA)平滑肌细胞膜电流的异同。方法:应用全细胞膜片钳技术研究SHR和Wistar大鼠BA平滑肌细胞在电流密度、电流组成以及自发性瞬时外向K+电流(STOCs)特性的异同。结果:①当指令电压为0、+20、+40和+60mV时,SHR与Wistar大鼠BA平滑肌细胞间电流密度存在统计学差异(P<0.01)。②SHR与Wistar大鼠BA平滑肌细胞膜电流都对1 mmol/L电压依赖的K+通道(Kv)阻断剂4AP和1 mmol/L大电导Ca2+激活K+通道(BKCa)阻断剂TEA敏感。③SHR的STOCs发放频率和电流幅度都远大于Wistar大鼠。1 mmol/LTEA基本完全阻断STOCs通道电流,而4-AP对STOCs没有影响。结论:SHR和Wistar大鼠脑动脉平滑肌细胞的电流密度存在差异,两种平滑肌细胞外向电流都由BKCa和Kv通道组成。SHR大鼠平滑肌细胞更易诱发由BKCa通道介导的STOCs。     

Exposure to stress differential vascular adaptive response in spontaneously hypertensive and Wistar rats: Role of nitric oxide, and prehypertensive and hypertensive states

Stress-induced vascular adaptive response in SHR was investigated, focusing on the endothelium. Noradrenaline responses were studied in intact and denuded aortas from 6-week-old (prehypertensive) and 14-week-old (hypertensive) SHR and age-matched Wistar rats submitted or not to acute stress (20-min swimming and 1-h immobilization 25 min apart), preceded or not by chronic stress (2 sessions 2 days apart of 1-h day immobilization for 5-consecutive days). Stress did not alter the reactivity of denuded aorta. Moreover, no alteration in the EC50 values was observed after stress exposure. In intact aortas, acute stress-induced hyporeactivity to noradrenaline similar between strains at both age. Chronic stress potentiated this adaptive response in 6- and 14-week-old Wistar but not in 6-week-old SHR, and did not alter the reactivity of 14-week-old SHR. Maximum response (g) in intact aortas [6-week-old: Wistar 3.25+/-0.12, Wistar/acute 1.95+/-0.12*, Wistar/chronic 1.36+/-0.21*(+), SHR 1.75+/-0.11, SHR/acute 0.88+/-0.08*, SHR/chronic 0.85+/-0.05*; 14-week-old: Wistar 3.83+/-0.13, Wistar/acute 2.72+/-0.13*, Wistar/chronic 1.91+/-0.19*(+), SHR 4.03+/-0.17, SHR/acute 2.26+/-0.12*, SHR/chronic 4.10+/-0.23; inside the same strain: *P < 0.05 relate to non-stressed rat, +P < 0.05 related to acute stressed rat; n = 6-18]. Independent of age and strain, L-NAME and endothelium removal abolished the stress-induced aorta hyporeactivity. CONCLUSION: The vascular adaptive response to stress is impaired in SHR, independently of the hypertensive state. Moreover, this vascular adaptive response is characterized by endothelial nitric oxide-system hyperactivity in both strains.     

Pressor responsiveness to vasopressin in spontaneously hypertensive rats

The present study was designed to find out whether pressor responsiveness to vasopressin (AVP) is altered in spontaneously hypertensive rats (SHR) in comparison with their normotensive controls (WKY). Blood pressure and heart rate changes after injection of graded doses of 2.5, 5.0 and 10.0 ng of AVP (Calbiochem) i.v. were compared in 9 conscious, unrestrained spontaneously hypertensive (SHR) and 11 normotensive Wistar Kyoto (WKY) rats, chronically instrumented with venous and arterial catheters. The threshold dose necessary to elicit a significant increase in blood pressure and reduction of heart rate was lower in WKY than in SHR. At each dose level the blood pressure elevation persisted for a longer period in WKY than in SHR. Bradycardia was greater in WKY than in SHR both in absolute terms and in relation to the blood pressure increase. Thus, the results reveal diminished pressor responsiveness to moderate doses of AVP in SHR in spite of suppressed reflex bradycardia. It is suggested that the peripheral action of AVP on the vascular system is attenuated in SHR.     

Breathing pattern of spontaneously hypertensive rats

The trachea of rats anaesthetized with sodium pentobarbitone was cannulated and the air flow velocity and the pressure of the oesophagus were measured. In the spontaneously hypertensive rats the breathing frequency was higher, the tidal volume and the effective lung resistance were smaller than that of the normotensive Wistar rats. It seems that the neurohumoral control of respiration in SHR animals differs from that of normotensive rats.