Small cell lung cancer (SCLC) demonstrates a strong etiological association with smoking. Although cigarette smoke is a mixture of about 4,000 compounds, nicotine is the addictive component of cigarette smoke. Several convergent studies have shown that nicotine promotes angiogenesis in lung cancers via the α7-nicotinic acetylcholine receptor (α7-nAChR) on endothelial cells. Therefore, we conjectured that α7-nAChR antagonists may attenuate nicotine-induced angiogenesis and be useful for the treatment of human SCLC. For the first time, our study explores the anti-angiogenic activity of MG624, a small-molecule α7-nAChR antagonist, in several experimental models of angiogenesis. We observed that MG624 potently suppressed the proliferation of primary human microvascular endothelial cells of the lung (HMEC-Ls). Furthermore, MG624 displayed robust anti-angiogenic activity in the Matrigel, rat aortic ring and rat retinal explant assays. The anti-angiogenic activity of MG624 was assessed by two in vivo models, namely the chicken chorioallantoic membrane model and the nude mice model. In both of these experimental models, MG624 inhibited angiogenesis of human SCLC tumors. Most importantly, the administration of MG624 was not associated with any toxic side effects, lethargy or discomfort in the mice. The anti-angiogenic activity of MG624 was mediated via the suppression of nicotine-induced FGF2 levels in HMEC-Ls. MG624 decreased nicotine-induced early growth response gene 1 (Egr-1) levels in HMEC-Ls, and reduced the levels of Egr-1 on the FGF2 promoter. Consequently, this process decreased FGF2 levels and angiogenesis. Our findings suggest that the anti-angiogenic effects of MG624 could be useful in anti-angiogenic therapy of human SCLCs.

SUMMARY Lerels and distribution of an angiotenslD-fonning enzyme, active at the physiological pH (isorenin), were determined in the central nervous system of 24 rats treated with 25 mg/kg of deoxycorticosterone (DOC) subcutaneously, twice a week, plus saline to drink during 30 days, and in 14 control animals. Different areas of the brain were excised and homogenized. Renin activity and concentration were determined by incubation of the supernatant of each homogenate at pH 7.2 alone and in the presence of an excess of renin substrate. The angiotensin generated was measured by radioimmunoassay. Concentration of the renin-like enzyme was significantly higher in the posterior hypophysis and in the brain stem of the experimental group; isorenin activity was higher in the hypothalamus, cerebral cortex, cerebellum, and brain stem of the DOC-salttreated rats than in the control rats. Changes in the angiotensin-forming enzyme in the central nervous system of experimental animals, active at physiological pH, suggest that this isoreoin system may play a role in the physiological response to DOC-salt in the rat. The significance of the brain isorenin system in the regulation of blood pressure requires further analysis.

The density of catecholamine-containing nerve fibers was studied in the cerebral and mesenteric arteries from normotensive Wistar-Kyoto rats (WKY), spontaneously hypertensive rats (SHR), and stroke-prone SHR (SHRSP) in the growing (SHR, WKY) and adult (SHR, SHRSP, WKY) animals. Cerebral arteries from SHR showed an increased adrenergic innervation from day 1. The nerve plexuses reached an adult pattern earlier in SHR than in WKY. The arteries from adult SHR and SHRSP (22 weeks old) showed a markedly higher nerve density than WKY. There was a positive linear correlation between blood pressure and nerve density for four cerebral arteries. The mesenteric arteries were not innervated at birth. However, hyperinnervation of these arteries in the SHR was already present at 10 days of age as compared with WKY. Sympathectomy with anti-nerve growth factor and guanethidine caused a complete disappearance of fluorescent fibers in the mesenteric arteries from SHR and WKY, and in the cerebral arteries of WKY. The same procedure caused only partial denervation of the cerebral arteries from hypertensive animals. We postulate that the increase in nerve density in the cerebral arteries from the hypertensive rats may contribute to the development of arterial hypertrophy in chronic hypertension through the trophic effect of the sympathetic innervation on vascular structure.

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SUMMARY Angiotensinogen is the most important component of the renin-angiotensin system present in the cerebrospinal fluid (CSF) of the rat. Its physiological significance as well as its origin have not been clearly elucidated. In this experiment we have examined plasma renin activity (PRA) and plasma and CSF angiotensinogen concentration under the following experimental conditions in male rats of the Wistar strain: 1) adrenalectomy (Adx) 4 days prior to sample collection; controls were sham Adx animals; 2) nephrectomy (Nx) 48 hours before blood and CSF collection; controls were sham Nx rats; 3) DOC-salt treatment (Cortexon depot, 50 mg/kg.s.c. twice a week) plus saline to drink was given during 4 weeks; controls were intact rats; 4) DOC-salt plus captopril: captopril (100 mg/kg/ day) in the drinking fluid was added to the treatment of experimental and control animals of Group 3; 5) two-kidney, two clip hypertension: silver clips placed in both renal arteries 8 weeks before samples collection; control: sham-operated rats; 6) water deprivation: rats deprived of water for 5 days; controls: intact rats; 7) peripheral sympathectomy: 6-hydroxydopamine (6-HODA) injected s.c. from birth until 16 weeks of age, adrenodemedullectomy and adrenal denervation performed at 8 weeks; controls were vehicle-injected animals.Determination of angiotensinogen concentration in plasma and CSF was accomplished by incubation of the samples with excess hog renin. The angiotensin I released as well as PRA were evaluated using an specific radioimmunoassay technique. PRA was significantly increased by Adx, captopril treatment, and water deprivation, and was almost suppressed by Nx, DOC-salt, and DOC-salt plus captopril treatment. Plasma angiotensinogen concentration was significantly elevated by Nx and significantly diminished by Adx, DOC-salt, and DOC-salt plus captopril, and captopril treatment. T HE presence of angiotensinogen in the brain and cerebrospinal fluid (CFS) of rabbits, dogs, rats, and humans has been clearly demonstrated. '^ Biochemical analysis has shown a similar molecular weight and electrophoretic mobility for plasma and CSF renin substrate; moreover, the final product released by both prohormones was identical to synthetic angiotensin I (AI) with respect to isoelectric point or pressor response.3 Clear differences between CSF and plasma angiotensinogen were also observed. In this regard, central renin substrate was heterogeneous with respect to isoionic points; the plasma protein presented only one component. In addition, im- munological differences between the central and peripheral renin substrate were also established.5 Although not definitively demonstrated, most of the available information supports a central origin for the angiotensinogen present in the brain and CSF of mammals and humans. Some experimental conditions determine a similar change in peripheral and central prohormone. 6 However, in a previous report 7 we found a clear dissociation between plasma and brain angiotensinogen in DOC-salt-treated hypertensi...