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Morphological Study of Gastric Lesions Developing in the Rat Under Several Damaging Conditions: Modifications Induced by Pretreatment with Zinc Acexamate

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Morphological Study of Gastric Lesions Developing in the Rat Under Several Damaging Conditions: Modifications Induced by Pretreatment with Zinc Acexamate Scanning Microscopy Volume 6 Number 3 Article 20 9-9-1992 Morphological Study of Gastric Lesions Developing in the Rat Under Several Damaging Conditions: Modifications Induced by Pretreatment with Zinc Acexamate M. Luisa Bravo Laboratorios Viñas, Barcelona Ginés Escolar Hospital Clínico, Barcelona Carmen Navarro Laboratorios Viñas, Barcelona Ramón Fontarnau Universidad de Barcelona Oriol Bulbena Laboratorios Viñas, Barcelona Follow this and additional works at: https://digitalcommons.usu.edu/microscopy Part of the Biology Commons Recommended Citation Bravo, M. Luisa; Escolar, Ginés; Navarro, Carmen; Fontarnau, Ramón; and Bulbena, Oriol (1992) "Morphological Study of Gastric Lesions Developing in the Rat Under Several Damaging Conditions: Modifications Induced yb Pretreatment with Zinc Acexamate," Scanning Microscopy: Vol. 6 : No. 3 , Article 20. Available at: https://digitalcommons.usu.edu/microscopy/vol6/iss3/20 This Article is brought to you for free and open access by the Western Dairy Center at DigitalCommons@USU. It has been accepted for inclusion in Scanning Microscopy by an authorized administrator of DigitalCommons@USU. For more information, please contact [email protected]. Scanning Microscopy, Vol. 6, No. 3, 1992 (Pages 855-864) 0891- 7035/92$5. 00 +. 00 Scanning Microscopy International, Chicago (AMF O'Hare), IL 60666 USA MORPHOLOGICAL STUDY OF GASTRIC LESIONS DEVELOPING IN THE RAT UNDER SEVERAL DAMAGING CONDITIONS: MODIFICATIONS INDUCED BY PRETREATMENT WITH ZINC ACEXAMATE M. Luisa Bravo, Gines Escolara, Carmen Navarro, Ram6n Fontarnaub, Oriol Bulbena • Departamento de Farmacologfa, Laboratorios Vifias, S.A., Barcelona, SPAIN aPresent address: Servicio de Hemoterapia y Hemostasia, Hospital Clfnico, Barcelona, SPAIN bPresent address: Servicio de Microscopfa Electr6nica, Universidad de Barcelona, Barcelona, SPAIN (Received for publication April 15, 1992, and in revised form September 9, 1992) Abstract Introduction Lesions developing in the gastric mucosa of the Development of gastric mucosa! damage induced rat after exposure to different gastric damaging agents by a variety of ulcerogenic agents has been studied in (100 mg/kg aspirin, and 70% or 100% ethanol) were as­ different experimental models [20]. The macroscopical sessed by scanning electron microscopy. The severity of quantification and morphological assessment of these the lesions was quantified according to morphological gastric injuries have contributed to the understanding of criteria. Modifications in the severity of these lesions the mechanisms involved in experimentally induced le­ induced by pretreatment with zinc acexamate were also sions which, in turn, has facilitated the development of analyzed. The scanning electron microscope revealed pharmacologic therapies for the human pathological that with the exception of absolute ethanol, which caused condition. distinctive morphological features, lesions found under To date, macroscopical criteria used to quantify the different experimental agents shared a common pat­ experimental lesions include measurement of the ulcer tern of progression. Ultrastructural lesions on surface length by morphometry [8] and the use of arbitrary epithelial cells preceded further alterations of parietal scales [l, II]. The light microscope allows the deter­ cells. After the integrity of the epithelial cells was lost, mination of the depth of the injury and morphological detachment of the parietal cells occurred, probably, modifications in the gastric mucosa. Unfortunately, ob­ through peptic digestion of the connections between cells servations in the light microscope are often restricted to and their extracellular matrices. Pretreatment of animals a limited portion of the stomach and fine ultrastructural with zinc acexamate increased the presence of mucus on changes taking place on the gastric surface might be the gastric surface and significantly prevented the pro­ overlooked. The scanning electron microscope (SEM), gression of lesions towards the severest stages. Ultra­ initially introduced for studies on gastric topology [21, structural damage of surface epithelial cells was not in­ 22], permits investigation of gastric lesions at ultrastruc­ fluenced by this treatment, but detachment of damaged tural level [4, 15, 17, 19, 3S]. The possibility of ac­ cells was clearly diminished. These data confirm the cessing different areas of the same sample, the great protective effect of zinc acexamate against gastric ag­ depth of field, and the three-dimensional viewing of the gressions. Moreover, our studies confirm the notion surfaces provide detailed structural information that is of that mucus secretion and maintenance of continuity on great help in reconstructing the natural progression of the gastric lumen by surface epithelial cells is of critical the lesions. Morphometric evaluation by SEM is cer­ importance in preventing the gastric damage induced in tainly limited [25], though some authors have developed these experimental models. morphological criteria that can be used to stage the severity of gastric lesions [34]. Key Words: Scanning electron microscopy, light Zinc acexamate (ZAC) has been demonstrated to microscopy, gastrotoxic agents, rat gastric mucosa, prevent the development of experimental gastric lesions gastric glands, surface epithelial cells, parietal cells, [9, 10, 12]. The protective effect of ZAC seems to re­ aspirin, ethanol. sult from its combined action on aggressive and defen­ sive mechanisms at the gastric mucosa! level. ZAC im­ • Address for Correspondence: proves gastric mucosa! defense preventing the disruption Oriol Bulbena, of the mucosa! barrier [ 11, 12], stabilizing biological Departamento de Farmacologfa membranes [24], and improving gastric microcirculation Laboratorios Vifias, S.A. [6] . This zinc compound has shown additional inhibitory C/Torrente Vidalet, 29 effects on pepsin and acid secretion [7]. A recent E-08012 Barcelona , SPAIN. investigation on ultrastructural gastric damage induced Phone No .: 34-3-2134700 by aspirin revealed that ZAC was capable of preventing 855 M.L. Bravo, G. Escolar , C. Navarro, et al. formation of deep erosions, but did not completely pre ­ Figure 1 (on the facing page). Scanning electron mi­ vent ultrastructural damage of surface epithelial cells crographs of rat gastric surfaces : (A) Normal appear­ [5]. ance of the undamaged gastric mucosa! surface in a con­ In the present study, SEM was used to investigate trol animal showing the smoothly undulated epithelial morphological changes taking place in the gastric mu­ surface layer delimiting the openings of gastric glands cosa of the rat subjected to different damaging agents and presence of the mucus layer (arrow). Treatment which are known to result in the development of severe with aspirin caused the disappearance of the mucus layer gastric ulcerations. Severity of the experimental lesions and induced a wide range of lesions often in the same was quantified according to morphological criteria. animal. (B) Focal erosions of the mucosa! surface Modifications in the severity of these lesions induced by progressing towards the mucous neck cells. (C) Inter­ pretreatment with ZAC were also analyzed. cellular junctions are preserved (arrow). (D) Destruc­ tion of the interfoveolar surface epithelial cells with Methods exposure of the architectural organization of the gastric glands (arrow). (E) Shedding of mucous and parietal General cells from the basal lamina. Membranes of these cells Female Wistar rats of similar weight (200 ± 10 appear well preserved. (F) Complex image of severely grams) and age (8-9 weeks old) were used [13]. Ani­ damaged mucosa with marked exfoliation, presence of mals were distributed in 4 groups consisting of 20 ani­ blood cell elements and fibrous material. Bar = 20 µm. mals each. One of these groups was used as a control and the remaining three groups were subjected to experi­ mental gastric damage according to the methods de­ scribed below. Animals were deprived of food but al­ the critical point method, and coated with gold up to a lowed free access to water during the 24 hours that pre­ thickness of 400 A in a sputter coating unit (Polaron ceded the induction of gastric damage. Half of the ani­ E5000). Specimens were coded so that observers were mals of each experimental group received a pretreatment unaware of the treatments. Observations were per­ with 100 mg/kg ZAC (p.o.) 60 minutes before exposure formed in a Hitachi S2300 SEM operated between 15 to the correspondent gastric aggressive agent. and 20 kV . Experimental models of gastric lesion Semi-quantitative morphometric analysis Acetylsalicylic acid (ASA): Gastric damage was The fundic mucosa of each stomach was systemat­ induced by oral administration of 100 mg/kg ASA . ically examined at low and high magnification in the Animals were sacrificed l hour later [5]. SEM . A progressive score of gastric damage from O (no Ethanol: To induce gastric lesions, 1 ml of lesion) to 5 (severest damage) was established . This 100% or 1 ml of 70% ethanol was administered p.o . scoring system is similar to that used by Winters et al. Animals were sacrificed 1 hour later [26]. [34] with minor modifications : Sacrifice and sampling 0. Gastric surface coated with mucus and nor- Animals were killed by cervical dislocation. The mal morphology of the epithelial cells in those areas stomachs were reached through medial laparotomy and where they are visible . an incision was made in the duodenum. A polytene can­ 1. Ultrastructural damage
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