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R M Lee

Publications and source records attributed to R M Lee.

At least 19 recordsLinked to original sources
PubMed1992

Evidence that extracellular signal-regulated kinases are the insulin-activated Raf-1 kinase kinases.

The Raf-1 proto-oncogene protein kinase can be phosphorylated and activated after stimulation of cells with insulin and a variety of other growth factors and mitogens. We recently presented evidence that insulin and certain other growth factors activated one or more Raf-1 kinase kinase activities (Lee, R.M., Rapp, U. R., and Blackshear, P.J. (1991) J. Biol. Chem. 266, 10351-10357). In the present study, four peaks of Raf-1 kinase kinase activity were identified after anion-exchange chromatography of cell lysates, and two of these were activated by insulin. Further chromatographic characterization of these two peaks of insulin-activated kinase activity indicated that they contained three apparently distinct kinase activities. Two of these activities comigrated with immunoreactive extracellular signal-regulated kinases (ERK) 1 and 2 (mitogen-activated protein kinase) through three different chromatographic separations. Both ERK1 and ERK2 phosphorylated Raf-1 with reasonably high affinity (Km for ERK1 = 90 nM; Km for ERK2 = 120 nM), and produced similar, complex phosphopeptide maps; both kinases also phosphorylated myelin basic protein. The third kinase activity also phosphorylated Raf-1 and myelin basic protein but did not comigrate exactly with either immunoreactive ERK1 or ERK2. We conclude that two and possibly three insulin-activated Raf-1 kinase kinases are members of the ERK family.

3T3 Cells↗
PubMed1992

Comparative effects of pravastatin and lovastatin on nighttime sleep and daytime performance.

Pravastatin and lovastatin, two HMG-CoA reductase inhibitors with similar cholesterol-lowering effects, differ in their lipid solubility. The hydrophilic characteristics of pravastatin may explain why the drug has not been detected in cerebrospinal fluid. On the other hand, lovastatin, a lipophilic compound, has been detected in the cerebrospinal fluid. Previous reports have suggested that lovastatin administration may be associated with insomnia, which reflects an action in the central nervous system. The effects of the two drugs on nocturnal sleep and day-time performance in young, healthy men have been assessed in randomized, double-blind, placebo-controlled studies. Computer-based performance tests were administered on two consecutive days before drug administration and at the end of a 3-week active drug or placebo treatment period. Results from both sites were combined for analysis. Neither pravastatin nor lovastatin significantly affected nocturnal sleep or daytime sleepiness in this study population, but lovastatin significantly affected daytime performance. In subjects treated with lovastatin, the results showed that two measures of performance, divided attention (p less than 0.05) and vigilance (p less than 0.01), worsened significantly from baseline as did global performance (p less than 0.01). Performance was not affected in the pravastatin and placebo groups. These results provide preliminary evidence of an adverse effect of lovastatin on daytime performance.

Adolescent↗
PubMed1992

Morphometric study of cerebral arteries from spontaneously hypertensive and stroke-prone spontaneously hypertensive rats.

OBJECTIVE: The importance of sympathetic innervation for the development of structural changes in the cerebral arteries of hypertensive animals was studied. DESIGN: Sympathetic denervation was induced with combined treatment from birth of antibody against nerve growth factor and guanethidine. Previous studies from our laboratory showed that this procedure not only caused a permanent denervation of the mesenteric arteries, but also prevented the development of hypertension in spontaneously hypertensive rats (SHR). METHODS: Morphometric measurement of the structural changes was carried out in the basilar, superior cerebellar, posterior cerebral and middle cerebral arteries from 28-week-old SHR, stroke-prone SHR, and normotensive Wistar-Kyoto rats. The results were compared with those obtained from cerebral arteries of sympathectomized rats. RESULTS: Total vascular wall cross-sectional area was significantly larger in the basilar and superior cerebellar arteries from hypertensive rats compared with normotensives. The change was characterized by an increase in the number of smooth muscle cell layers. There were also differences between the two hypertensive groups in some arteries. Sympathetic denervation attenuated the development of hypertension and vascular changes in some arteries. There was a positive linear correlation between blood pressure and medial cross-sectional area, and between blood pressure and the number of smooth muscle cell layers for the four arteries analysed. CONCLUSION: Sympathetic nerves have a trophic influence upon the remodelling of some cerebral arteries during the development of genetic hypertension.

Animals↗
PubMed1992

The effect of neonatal treatment of rats with nerve growth factor on the blood pressure and structure of the mesenteric arteries.

Newborn male Wistar rats were treated with nerve growth factor daily by subcutaneous injection for 2 weeks, and control rats were treated with either cytochrome c or buffered saline. Average body weight of the treated animals was lower than that of the controls during the 2 weeks of treatment, but became similar to that of the controls thereafter. Tissue levels of norepinephrine were elevated in the brain, adrenal glands, mesenteric arteries, and vas deferens of the treated animals immediately after the treatment, but became similar in the three groups 2 weeks after the termination of the treatment. Blood pressure and heart rate were measured beginning at 4 weeks of age until 28 weeks, when the rats were sacrificed and the mesenteric arteries sampled for morphometric measurements of vessel wall dimensions. Pretreatment with nerve growth factor did not affect blood pressure, nor heart rate. Structural alteration of the three types of mesenteric arteries was also absent in the treated animals. We conclude that even though neonatal treatment of normal Wistar rats with nerve growth factor for 2 weeks induced an elevation of the norepinephrine levels in several tissues at the end of the treatment period, it was not sufficient to produce hypertension and structural alterations in the blood vessels.

Aging↗
PubMed1992

Influence of chronic nadolol treatment on blood pressure and vascular changes in spontaneously hypertensive rats.

Chronic treatment of spontaneously hypertensive rats (SHR) and Kyoto-Wistar normotensive rats (WKY) with nadolol was carried out from gestation until 28 weeks of age. Nadolol treatment caused some lowering of blood pressure but did not prevent the development of hypertension or cardiac hypertrophy in the SHR, in spite of significant beta-blockade. The lumen of large mesenteric arteries from control SHR was smaller than from WKY, and nadolol treatment increased the lumen size in the SHR. An increased number of smooth muscle cell layers present in the control SHR as compared with WKY was reduced slightly by nadolol treatment. However, the changes produced by nadolol did not reach the levels of control and treated WKY. In the aorta, the incidence of polyploid smooth muscle cells was higher in the SHR than the WKY in the control group. Nadolol treatment reduced the percentage of polyploid cells in both SHR and WKY, so that the difference between these two groups of animals was eliminated in the treated groups. The tissue level of norepinephrine in the plasma, heart, mesenteric arteries, and adrenal glands in the SHR and WKY was not affected by the treatment. We suggest that the ineffectiveness of nadolol in preventing hypertension development may be due to its lack of effect in preventing primary changes in the resistance arteries, and that the development of polyploidy of smooth muscle cells may be mediated by beta-receptors.

Adrenal Glands↗
PubMed1992

Incidence of multinucleated and polyploid aortic smooth muscle cells cultured from different age groups of spontaneously hypertensive rats.

Cell size and incidence of multinucleated, polyploid cells in cultured aortic smooth muscle cells from different age groups of spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto rats (WKY) were compared. Smooth muscle cells from SHR were generally larger than those from WKY, and the percentage of multinucleated smooth muscle cells was always higher in SHR than WKY in the three age groups of rats studied (3-4, 10-12, and 28-30 weeks). In smooth muscle cells from the 3- to 4-week group, there was a positive correlation between cell diameter and the percentage of multinucleated smooth muscle cells. Microdensitometric measurements also showed that the incidence of polyploid smooth muscle cells was always higher in SHR than WKY in the three age groups. There was a positive correlation between DNA density and nuclear area measurements in all the age groups of SHR and WKY. We conclude that cultured aortic smooth muscle cells from different age groups of SHR and WKY contained heterogeneous populations of cells and that, under our culture conditions, the polyploidy of the smooth muscle cells found in vivo was maintained in the SHR and WKY.

Aging↗
PubMed1992

Antihypertensive effect of gamma-linolenic acid in spontaneously hypertensive rats.

The effects of chronic treatments of adult (aged 16-17 weeks) spontaneously hypertensive rats (SHRs) with different doses of gamma-linolenic acid (GLA) on blood pressure, heart rate, and body weight were studied. Twice-daily injection of SHRs with GLA lowered systolic blood pressure from 175 +/- 4 to 145 +/- 4 mm Hg within 1 week; systolic blood pressure in all three treated groups became stabilized in the normotensive range after 2 weeks of treatment. Control SHRs injected with olive oil showed only a transient decrease in systolic blood pressure on the third day. Heart rate and body weight were not affected by GLA treatment. Withdrawal of GLA treatment resulted in a rapid rise in systolic blood pressure within 1 day from 140 +/- 3 to 165 +/- 3 mm Hg, and it stabilized after 1 week at 191 +/- 5 mm Hg in the three experimental groups. A rapid increase in systolic blood pressure from 175 +/- 5 to 203 +/- 5 mm Hg was also observed in the control group treated with olive oil 1 day after the withdrawal of the treatment. Addition of aspirin (3 mg/kg) with the GLA treatment in olive oil abolished the antihypertensive effect of GLA. In contrast, once-daily treatment with GLA also lowered systolic blood pressure of the SHR, but blood pressure was still in the hypertensive range (170 +/- 6 mm Hg). Systolic blood pressure of control SHRs treated with olive oil was not affected. Plasma from untreated SHRs contained a small amount of GLA. One hour after the injection, the plasma level of GLA increased. We conclude that GLA when given twice daily is an effective antihypertensive agent in the SHR.

Animals↗
PubMed1992

Adenosine metabolism in small coronary arteries of pig.

Adenosine preferentially relaxes small coronary arteries over large ones, and small bovine coronary arteries are also known to have a higher density of adenosine receptors. Here we report a possible role of adenosine metabolism in this process. Subcellular fractions, from right coronary artery (lumen diameter of 2-3 mm) of pig designated as large coronary artery and its subsequent branches (lumen diameter of 0.5-1 mm) as small coronary arteries, were prepared and characterized. In comparison to the various large artery subcellular fractions, the corresponding small artery fractions were richer in 5'-nucleotidase but poorer in adenosine deaminase. Thus a cascade of events may promote adenosine relaxation in small coronary arteries: higher activity of 5'-nucleotidase leads to production of more adenosine, larger number of receptors allows greater reactivity to adenosine, and lower adenosine deaminase level promotes prolonged action of adenosine.

5'-Nucleotidase↗
PubMed1991

Evidence for one or more Raf-1 kinase kinase(s) activated by insulin and polypeptide growth factors.

The protein product of the Raf-1 proto-oncogene is a protein serine/threonine kinase that is activated after stimulation of cells with insulin and other mitogens. To investigate the mechanism of this activation, we used purified Raf-1 expressed in E. coli as a substrate for a putative Raf-1 protein kinase kinase. In three different insulin-sensitive cell types, insulin activated Raf-1 kinase kinase activity in crude cytosolic cellular fractions. The insulin stimulation of this activity was evident as early as 2 min after exposure to insulin, maximal at 5-8 min, and inapparent at 15 min. Phosphoamino acid analysis of phosphorylated Raf-1 revealed that serine was the primary phosphate acceptor for the insulin-activated kinase or kinases; small amounts of phosphothreonine were also detected. The insulin effect occurred in cells depleted of protein kinase C, and in extracts depleted of endogenous Raf-1 kinase by immunodepletion; these data argue against protein kinase C or Raf-1 kinase itself being the insulin-stimulated activity. The insulin-activated kinase or kinases phosphorylated the Raf-1 protein on multiple sites in vitro, as evidenced by tryptic mapping; at least some of these appeared to overlap with sites phosphorylated in response to serum in intact cells. Several other mitogens and growth factors stimulated Raf-1 kinase kinase activity, including epidermal growth factor, platelet-derived growth factor, fibroblast growth factor, serum, and phorbol 12-myristate 13-acetate. This insulin- and mitogen-stimulated Raf-1 kinase kinase activity may play a role in mediating the phosphorylation and possibly the activation of the Raf-1 kinase by insulin and other growth factors.

Animals↗
PubMed1991

Interaction between sympathetic nervous system and adrenal medulla in the control of cardiovascular changes in hypertension.

The role of the sympathetic nervous system and adrenal medulla in the development of cardiovascular changes and hypertension was studied in spontaneously hypertensive rats (SHRs), and the results compared with age-matched normotensive Wistar-Kyoto (WKY) rats. Sympathectomy was initiated in newborn rats through daily injection with antiserum to nerve growth factor for 1 week, followed by daily injection with guanethidine for 3 weeks. Removal of the adrenal medulla was carried out in 4-week-old rats after the last guanethidine injection. Such a combination treatment was effective in permanently preventing the development of hypertension in the SHRs, and the blood pressure was maintained at the level of WKY rats. The heart rate of the SHRs and WKY rats was not affected by such treatment. Hypertrophy of the heart and of the vessel wall in the mesenteric arteries was also prevented by such treatment. We conclude that in the SHR, the sympathetic nervous system and the adrenal medulla are essential for the development of cardiovascular changes and hypertension.

Adrenal Medulla↗
PubMed1991

Opposite effects of arterial vasodilators on cardiac vs. arterial hypertrophy and sympathetic activity in spontaneously hypertensive rats.

To investigate whether cardiac and arterial structure and sympathetic activity changes in a similar fashion during chronic arterial vasodilation, we evaluated the morphology and sympathetic activity of the mesenteric arterial bed and the left (LV) and right (RV) ventricles of 16-week-old Wistar-Kyoto (WKY) and spontaneously hypertensive rats (SHRs) after 35 and 70 days of treatment with the arterial vasodilator minoxidil. The minoxidil-induced antihypertensive response was associated with an increase in ventricular sympathetic activity, potentiation of RV hypertrophy (RVH), and the development of eccentric LV hypertrophy (LVH). In the mesenteric arterial bed, minoxidil decreased the sympathetic activity, increased the lumen of the superior mesenteric artery, and decreased the medial area of the large and small mesenteric arteries. We conclude that the contrasting effects of minoxidil on cardiac vs. arterial structure may--in part--relate to selective effects on regional sympathetic activity.

Animals↗
PubMed1991

Arterial vasodilation and cardiovascular structural changes in normotensive rats.

In normotensive rats, the arterial vasodilator minoxidil causes right ventricular hypertrophy (RVH) and eccentric left ventricular hypertrophy (LVH). To assess whether this trophic effect of minoxidil extends to the vasculature and to examine possible mechanisms involved, alterations in cardiac and arterial (superior, large and small mesenteric arteries, carotid and basilar arteries) structure were evaluated in relation to changes in indexes of cardiac volume load and cardiac and arterial sympathetic activity during long-term (35 and 70 days) treatment of normotensive rats with minoxidil alone or in combination with the diuretic hydrochlorothiazide (HCTZ). Minoxidil alone increased LV and RV weights, LV internal diameter, and medial area of the superior mesenteric artery but did not affect any of the other arteries evaluated. When combined with HCTZ, long-term minoxidil caused concentric LVH rather than eccentric LVH and no longer increased the medial area of the superior mesenteric artery. Neither treatment had any persistent effect on blood pressure, heart rate, or plasma catecholamines. However, minoxidil significantly increased cardiac and arterial (superior and large mesenteric artery) norepinephrine turnover rates, cardiac filling pressures, and plasma and blood volumes. When combined with HCTZ, short-term (1 wk) minoxidil still increased cardiac filling pressures. However, intravascular volume expansion during chronic treatment was significantly attenuated. These results suggest that chronic cardiac volume load appears to determine the type of cardiac hypertrophy induced by a nonhemodynamic mechanism (possibly cardiac sympathetic activity) activated by minoxidil. Intravascular volume expansion or increased arterial flow appears to be responsible for medial hypertrophy of the superior mesenteric artery, but absence of a trophic response in other arteries suggests that another, local mechanism contributes.

Animals↗
PubMed1991

Prevention of hypertension and vascular changes by captopril treatment.

Treatment of female spontaneously hypertensive rats (SHR) and control Wistar-Kyoto (WKY) rats with captopril was carried out by the addition of the drug in the drinking water throughout pregnancy and lactation and after weaning. At 28 weeks of age, average systolic blood pressure of treated SHR was 113 +/- 3 mm Hg, which was below that of control SHR (188 +/- 3 mm Hg) and WKY rats (124 +/- 3 mm Hg). Body weight and heart rate of the SHR were not affected by the treatment. Tissue level of catecholamines was increased by captopril treatment in the superior cervical ganglia but remained unchanged in the plasma, heart, mesenteric arteries, and the adrenal glands of both SHR and WKY rats. Left ventricular weight, wall thickness, and internal diameter of the left ventricle in the SHR were reduced by the treatment. Morphometric measurements of the mesenteric arteries showed that vascular alterations present in the control SHR were prevented by the treatment. In the superior mesenteric artery and large mesenteric artery, smaller lumen size at maximal relaxation found in the control SHR was normalized to the level of the WKY rats. Hypertrophy of the medial wall in the superior mesenteric, large and small mesenteric arteries, and an increase in the number of smooth muscle cell layers in the large mesenteric artery of the SHR were prevented by the treatment. Perfusion study of the mesenteric vascular bed showed that reactivity of these vessels to norepinephrine was reduced, and sensitivity to norepinephrine (as determined by the effective dose that causes 50% of maximal response) was increased in the SHR by captopril treatment. Sensitivity of the tail artery in response to norepinephrine was not altered by the treatment. We conclude that long-term treatment with captopril of SHR before and after birth prevented the development of hypertension, structural and functional alterations of the mesenteric arteries, and cardiac hypertrophy.

Animals↗
PubMed1991

Effect of hydralazine on the mesenteric vasculature of hypertensive rats.

To test whether structural alterations observed in the mesenteric vasculature of Wistar-Kyoto spontaneously hypertensive rats (SHR) were dependent on the presence of hypertension, male SHR and Wistar-Kyoto normotensive (WKY) rats were treated in utero and postnatally with hydralazine up to 28 weeks of age. Treated SHR, WKY, and untreated WKY rats had comparable blood pressures that were less than those of untreated SHR. Treatment altered the dimensions of the superior mesenteric, intermediate-sized, and small arteries of the mesenteric vasculature. In the case of the superior mesenteric artery and intermediate vessels, hydralazine treatment increased the lumen and medial cross-sectional areas of the arteries in WKY rats and slightly decreased both parameters in SHR. Within the small arteries, treatment significantly increased the lumen size in SHR but not WKY rats and had no significant effect on the media of the vessels. Despite the above alterations, the media-to-lumen cross-sectional area ratios remained significantly elevated in SHR over WKY rats in both the treated and control groups of animals within all classes of arteries. The results indicate that there is an inherent increase in the quantity of media surrounding the arteries of SHR when compared with WKY rats that cannot be abolished by normalizing the blood pressure in utero and postnatally with hydralazine treatment. In SHR, such changes persist not only in arteries that exhibit an increase in the media-to-lumen ratio before hypertension but also in the superior mesenteric artery in which an increase in the ratio occurs after hypertension development.

Animals↗
PubMed1991

Combined effect of neonatal sympathectomy and adrenal demedullation on blood pressure and vascular changes in spontaneously hypertensive rats.

Neonatal sympathectomy using a combined treatment with antiserum to nerve growth factor and guanethidine during the first 4 weeks after birth was carried out in spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto (WKY) rats. Bilateral adrenal demedullation was performed in 4-week-old sympathectomized SHR and WKY rats. The development of hypertension in SHR was prevented by sympathectomy, but the blood pressure (BP) was still higher than in age-matched WKY rats. Demedullation reduced the BP of sympathectomized SHR to the same level as that of WKY rats. Heart rates of SHR and WKY rats were not affected by the treatments. Morphometric measurements of the mesenteric arteries showed that sympathectomy significantly reduced the medial mass in the mesenteric arteries of SHR, mainly through a reduction in the number of smooth muscle cell layers. In sympathectomized SHR, demedullation increased the lumen size of muscular arteries under maximally relaxed conditions, which might explain the further reduction in BP in these animals. Demedullation in sympathectomized SHR and WKY rats caused a decrease in smooth muscle cell layers in the superior mesenteric artery, but the same treatment resulted in a slight increase in the number of smooth muscle cell layers in the large and small mesenteric arteries of SHR and WKY rats. Adventitial area was increased in some mesenteric arteries of SHR and WKY rats by sympathectomy, and demedullation caused a further increase in the size of adventitia in WKY rats. Heart weight in SHR was normalized to the level found in WKY rats by sympathectomy and demedullation. We conclude that in sympathectomized SHR, the elevated BP was maintained by the adrenal medulla.

Adrenal Medulla↗
PubMed1991

Sympatholytic interventions and vascular remodelling.

Most of the results on the effects of sympatholytic agents on vascular remodelling came from studies involving hypertensive animals. Therefore, it is often difficult to dissociate the antihypertensive property of the drugs from those caused by the direct action of the drugs on the structure of the blood vessels. However, some general conclusions can be drawn in some areas. Very little information is available on the effects of sympatholytic agents on the structure and function of the endothelial cells, and on the structure of the internal elastic lamina. Necrotic changes of the endothelial cells, and destruction of the internal elastic lamina present in hypertensive animals are prevented with sympatholytic agents, which may be related to the antihypertensive property of the drugs. The density of the fenestrations in the internal elastic lamina is decreased by sympathectomy. In the media, proliferation of vascular smooth muscle cells in growing animals is usually interfered with by sympatholytic drugs, so that the medial mass is normally smaller than that in the untreated control. In most animals, treatment of the animals with sympatholytic agents causes an increase in the biosynthesis, or accumulation of collagen in several types of arteries in the rats and rabbits. In addition to adrenergic nerves, the density of some peptidergic nerves is also reduced by these drugs. The effect of sympathectomy on lumen size is less clear, because lumen size was increased in some studies, and decreased in others. It is clear that remodelling of the vessel wall does occur in some instances, but in general, it is important to distinguish the growth retardation of the animals due to the treatment, from the effect which might be specific to the vessel wall.

Animals↗