Overexpression of cyclin E in Mongolian gerbil with Helicobacter pylori-induced gastric precancerosis

AIM: To explore dysregulation of cyclin E in malignancies,and to further investigate the role of cyclin E in Helicobacterpylori ( H. pylori)-induced gastric precancerosis.METHODS: Four-week-old specific pathogen-free maleMongolian gerbils were employed in the study. 0.5 mL 1 ×108 cfu@ L- 1 suspension of H. pylori NTCC11637 in Brucellabroth was inoculated orally into each of 20 Mongolian gerbils, and a further 20 gerbils were inoculated with Brucella brothas controls. 10 of the infected gerbils and 10 of the non-infected control gerbils were sacrificed at 25, 45 wk afterinfection. The expression of cyclin E was analyzed by RT-PCR and immunohistochemical studies with monoclonalantibody to cyclin E in Mongolian gerbil of H. pylori-induced gastric precancerosis.RESULTS: H. pylori was constantly detected in all infectedanimals throughout the study. At 25 wk after infection of H.pylori, ulcers were observed in the antral and body ofstomach ( n = 6). Histological examination showed that allanimals developed severe inflammation and multifocallymphoid follicles appeared in the lamina propria andsubmucosa of gastric antrum. At 45 wk after infection of H.pylori, severe atrophic gastritis (n = 10), intestinalmetaplasia (n = 8) and dysplasia (n = 6) could beobserved. Cyclin E mRNA levels were significantly more at25 wk after infection of H. pylori (1.27±0.26), and at45 wkafter infection of H. pylori (1.82 ± 0.39 ) than control-animals (0.59 ± 0.20, P＜ 0.01) ; cyclin E mRNA levels wereevaluated by 2.2-fold at 25 wk ( P ＜ 0.01 ) and 3. 1-fold at 45wk ( P ＜ 0.01 ) precancarosis induced by H. pylori, whencompared with control gastric epithelium of Mongoliangerbil. Immunohistochemical staining revealed exclusivenuclear staining of cyclin E. Furthermore, there wes asequential increase in cyclin E positive cells from normalepithelium to precancerosis.CONCLUSION: Overexpression of cyclin E occurs relativelyearly in gastric tumorigenesis in this model.

Overexpression of c-fos in Helicobacter pylori-induced gastric precancerosis of Mongolian gerbil

AIM: To explore dysregulation of c-fos in several human malignancies, and to further investigate the role of c-fos in Helicobacter pylori ( H. pylori)-induced gastric precancerosis.METHODS: Four-week-old male Mongolian gerbils were H. pyloriNCTC 11 637 in Brucella broth were inoculated orally into 20 Mongolian gerbils. Another 20 gerbils were inoculated with Brucella broth as controls. 10 of the infected gerbils and 10 of the non- infected control gerbils were sacrificed at 25 and 45 weeks after infection. The stomach of each gerbil was removed and opened for macroscopic observation. The expression of c-fos was analyzed by RTPCR and immunohistochemical studies in H. pylori-induced gastric precancerosis of Mongolian gerbil. Half of each gastric antrum mucosa was dissected for RNA isolation and RTPCR. β-actin was used as the housekeeping gene and amplified with c-fos as contrast. PCR products of c-fos were analyzed by gel image system and the level of c-fos was reflected with the ratio of c-fos/β-actin. The immunostaining for c-foswas conducted using monoclonal antibody of c-fosand the StreptAvidin-Biotin-enzyme Complex kit.RESULTS: H. pyloriwas constantly found in all infected animals in this study. After infection of H. Pylorifor 25 weeks,ulcers were observed in the antral and the body of stomach of 60 % infected animals (6/10). Histological examination showed that all animals developed severe inflammation, especially in the area close to ulcers, and multifocal lymphoid follicles appeared in the lamina propria and submucosa. After infection of H. Pylorifor 45 weeks, severe atrophic gastritis in all infected animals, intestinal metaplasia in 80 % infected animals (8/10) and dysplasia in 60 % infected animals (6/10) could be observed. C-fos mRNA levels were significantlyhigher after infection of H. pylorifor 25 weeks (1.84±0.79),and for 45 weeks (1.59±0.37) than those in control-animals (0.74±0.22, P＜0.01). C-fos mRNA levels were increased 2.5-fold by 25th week (P＜0.01) and 2.1-fold by 45th week (P＜0.01) in precancerosis induced by H. pylori, when compared with normal gastric epithelium of Mongolian gerbil. Immunohistochemical staining revealed exclusive nuclear staining of c-fos. Furthermore, there was a sequential increase in c-fos positive cells from normal epithelium to precancerosis.CONCLUSION: The study suggested that overexpression of c-fos occurs relatively early in gastric tumorigenesis in this precancerosis model induced by H, pylori.

Mechanisms of Apigenin-7-glucoside As a Hepatoprotective Agent

Ixeris chinesis (Thunb.) Ankai has been used as a Chinese folk medicine, but only scanty information is available on the physiological and biochemical functions of the compounds extracted from I. chinesis. In the present study the effects of apigenin -7-glucoside (APIG) isolated from I. chinesis against liver injury caused by carbon tetrachloride (CCl4) were investigated. Methods The contents of malondialdehyde (MDA), glutamic pyruvic transaminase (GPT), glutamic oxaloacetic transaminase (GOT), and reduced glutathione (GSH) were evaluated by spectrophotography. The content of 8-Hydroxydeoxyguanosine (8-OHdG) was measured with high-performance liquid chromatography (HPLC) equipped with electrochemical and UV detection methods. The antioxidant activity of APIG was evaluated using chemiluminescence single photon counting technology. Results CCl4 significantly increased the enzyme activities of GPT and GOT in blood serum, as well as the level of MDA and 8-OHdG in liver tissue, and decreased the levels of GSH. Pretreatment with APIG was able not only to suppress the elevation of GPT, GOT, MDA and 8-OHdG, and inhibit the reduction of GSH in a dose-dependent manner in vivo, but also to reduce the damage of hepatocytes in vitro. On the other hand, we also found that APIG had strong antioxidant activity against reactive oxygen species (ROS) in vitro in a concentration-dependent manner. Conclusion The hepatoprotective activity of APIG is possibly due to its antioxidant properties, acting as scavengers of ROS. These results obtained in vivo and in vitro suggest that APIG has protective effects against hepatic oxidative injury induced by chemicals. Further studies on the pharmaceutical functions and immunological responses of APIG may help its clinical application.