Studies on the Paneth Cell of the Mouse Ileum after Administration of Cyclophosphamide

Moon, Yong Suk; Kang, Jeong Soo; Kim, Bong Seon; Baek, Sun Yong; Huh, Eun Young; Hong, Hae Nam; Kim, Jae Bong

Abstract

This study was carried out on ICR mice, male, weighing about 26~35 g in order to investigate the effects of cyclophosphamide (CP) on the Paneth cells and their glycoconjugates. They were given intraperitoneally CP (Sigma, USA) 150 mg/kg body weight. Control mice were given as same amount of distilled water. The mice were sacrificed after 12 hours and on day 1, 2, 4, 6, 9 and 14 after CP injection. Sections were prepared from the region upper 1~2 cm from the end of the ilea. The material for histological examination was fixed in 10% buffered formalin. Some of the preparation 4 mm thick from the paraffin blocks stained with hematoxylin and eosin. The average numbers of the cells (Paneth cell index : PCI) were counted in the longitudinally sectioned 20 intestinal glands and semiquantitive granulation indices (Paneth cell granulation index : PGI) were obtained arithmatically weighted method to 3 cell types classified according to the degree of granularity of the cells. The other sections were incubated with 8 species of lectins (GS I B4, PSA, SBA, sWGA, UEA I, ECL, PNA and LFA). In order to increase the specificity of the reactions, the sections were applicated with ABC system. And then the sections were incubated DAB and were counterstained with hematoxylin. The results observed by light microcope were as follows. 1. The Paneth cell index (PCI) was 155.5 in control mice, while the PCI from the mice after 12 hours CP injection was 88.3. The Paneth cell granulation index (PGI) decreased from 316.0 in control mice to 152.3 in 12 hours after the CP administration. 2. The PCI increased to 141 and the PGI was 354 on day 2 after CP administration, which was higher in number than those of the control mice. It was characterized that the Paneth cells packed with numerous eosinophilic granules in the apical region increased in great numbers on day 2. 3. The PCI and PGI decreased on day 4 and day 5, and began to increase on day 9, which recovered to the similar level of the control mice. 4. Apoptotic-like cells increased suddenly in great numbers 12 hours after the CPA injection and began to decrease on day 1. Most of the dying cells seem to come from stem cells of the crypts and a small numbers of them from Paneth cells. 5. Paneth cells exhibited an extensive binding pattern for SBA, sWGA, and showed a restricted binding pattern for GS I B4 and UEA I. PSA, PNA, LFA. ECL showed negative reaction with Paneth cells. 6. It seems that Paneth cells can be classified according to the composition of the glycoconjugate in the granule and the stages of the cell maturation. The glycoconjugates in the halo is thought different from that in the core of the secretory granules. 7. The Paneth cell granules generally showed stronger reaction with the lectins in 12 hours after the CP admini-stration. 8. It is thought that the core of the granules decomposed earlier than the halo of the granules, and the granules of the cells reacted negatively with the lectins secreted earlier than those of the cells showed strong reaction with the lectins after the CP injection.