Duodenogastric Reflux and Helicobacter pylori Infection Synergistically Increase Gastric Mucosal Cell Proliferative Activity in Mongolian Gerbils

Background: Helicobacter pylori and duodenogastric reflux (DGR) are both recognized as aetiological factors in chronic gastritis and gastric carcinogenesis. In this study, a Mongolian gerbil (MG) model was used to investigate the histopathological changes in the gastric mucosa resulting from DGR and/or H. pylori infection. Methods: One-hundred-and-eleven 7-week-old, specific-pathogen-free, male MGs were divided into four groups: normal controls, gerbils with surgically induced DGR, and H. pylori-infected gerbils with and without DGR. Gerbils were killed 4, 12 and 26 weeks after DGR surgery, their stomachs removed and sections prepared. Sections were fixed immediately in 20% phosphate-buffered formalin and subjected to haematoxylin and eosin staining, Alcian blue at pH 2.5/periodic acid-Schiff staining, and immunostaining for smooth muscle cells, H. pylori and 5′-bromo-2′-deoxyuridine (BrdU). Results: The gastric mucosa of H. pylori-infected gerbils showed chronic active gastritis irrespective of DGR throughout the experimental period. The gastric mucosa of H. pylori-infected gerbils with DGR demonstrated higher BrdU labelling than in the other groups. Conclusions: In MGs, DGR and H. pylori infection synergistically increased gastric mucosal cell proliferative activity. DGR and H. pylori infection may be involved synergistically in gastric carcinogenesis by increasing cell proliferative activity.     

黄苔胃病患者胃粘膜凋亡基因及相关蛋白的表达

目的:探讨胃病患者黄苔与胃粘膜组织细胞凋亡之间的关系.方法:选取黄苔胃病患者62例,采用免疫组化法与63例白苔胃病患者进行对照,采用免疫组化法检测胃粘膜增殖细胞核抗原(PCNA)、p53、bcl-2、fas凋亡基因相关蛋白.结果:黄苔患者p53、fas表达均明显高于白苔患者(均P<0.05),而PCNA、bc1-2表达则与白苔患者差异均无统计学意义(均P>0.05).结论:黄苔胃病患者伴有促凋亡基因相关蛋白的过度表达,黄苔是胃内有活动性炎症的较灵敏的指标.     

三九胃泰颗粒对大鼠急性胃粘膜损伤的修复作用

目的 :观察三九胃泰颗粒对胃粘膜损伤修复和细胞增殖活性调控的影响。方法 :采用无水乙醇 1m l胃饲诱发急性胃粘膜损伤大鼠模型并用三九胃泰颗粒治疗 ,免疫组织化学技术检测胃上皮细胞的增殖改变 ,早期应答基因 c- jun和 c- met的表达。结果 :三九胃泰颗粒、胃舒颗粒、养胃冲剂及丽珠得乐治疗组大鼠胃粘膜损伤指数均显著低于无水乙醇模型组及自然恢复组 ,尤以三九胃泰颗粒治疗后为甚 (P <0 .0 5 ) ;免疫组织化学显示三九胃泰颗粒、胃舒颗粒、养胃冲剂及丽珠得乐治疗组大鼠胃粘膜增殖细胞核抗原标记的阳性细胞数量、c- jun和 c- met的阳性表达均有高于无水乙醇模型组及自然恢复组的趋势 ,尤以三九胃泰颗粒治疗后为甚 (P <0 .0 5 )。结论 :三九胃泰颗粒能促进急性胃粘膜损伤的修复     

胃溃灵对胃粘膜损伤大鼠胃粘膜分泌的影响

目的 :研究胃溃灵对乙酸引起胃粘膜损伤大鼠胃粘膜分泌的影响。方法 :将 5 0只 SD大鼠制成乙酸胃粘膜损伤模型 ,并随机将大鼠等分为 5组 ,次日起给每组大鼠分别灌服等量生理盐水、大、中、小剂量 ( 12 g/ kg、6g/kg、3 g/ kg)胃溃灵、雷尼替丁 ,10 d后处死大鼠 ,观察大鼠胃粘膜损伤程度。采用阿尔新蓝与胃液中糖蛋白结合的方法 ,分别测定大鼠胃内游离粘液量、胃壁粘液量。结果 :胃溃灵能明显提高大鼠胃内游离粘液、胃壁粘液的分泌量 ,能明显抑制乙酸对大鼠胃粘膜的损伤。结论 :胃粘液分泌量增加可加强对粘膜的屏障作用 ,这是胃溃灵保护胃粘膜损伤的机制之一。     

Gastric Mucosa Epithelial Cell Kinetics Are Differentiated by Anatomic Site and Helicobacter Pylori Infection

Changes in epithelial cell turnover related to Helicobacter pylori infection may contribute to gastric cancer development. The response of different anatomic sites of the gastric mucosa to H. pylori is not known. We studied apoptosis and cell proliferation at the grater and lesser curvature of the antrum and corpus, the fundus, and the cardia from 9 H. pylori gastritis patients and 11 H. pylori-negative controls with normal histology. Proliferation was highest at the major curve of the antrum and lowest at the fundus, and apoptosis was highest at the cardia and lowest at the major curve of the antrum in both H. pylori gastritis and normal mucosa. Proliferation was significantly higher at all anatomic sites, while apoptosis was significantly lower only at the major and lesser curve of the corpus in H. pylori gastritis compared with normal controls. Our data suggest that gastric mucosa epithelial cell kinetics is differentiated by the anatomic site and H. pylori infection.     

康胃方对沙土鼠幽门螺杆菌的根除及胃粘膜保护作用

目的：观察康胃方对沙土鼠幽门螺杆菌（Hp)根除及胃粘膜细胞保护作用。方法：蒙古沙土鼠42只,随机分为病理模型组、三联治疗组及康胃方高、低剂量组,每日灌胃给予Hp菌液,共3次。用脲呼吸试验（UBT）确认Hp感染情况并接近相应的治疗药物。另12只动物未予感染。治疗后第4周处死动物,测定胃粘膜活性氧（RO）、一氧化氮（NO）、诱生型一氧化氮合成酶（iNOS)、MnSOD及NPSH。结果：Hp感染各组动物,UBT与胃膜粘所含RO、NO、iNOS和MnSOD明显升高,但NPSH显著下降（P＜0．01）。经抗感染治疗后UBT值降低,其他指标明显改善（P＜0．01）。结论：康胃方对蒙古沙土鼠感染Hp有明显的根除作用,并通过改善感染动物胃粘膜某些生化成分的含量或活性,发挥其细胞保护作用。     

Effect of Cimetidine on Gastric Mucosal Cell Proliferation in Man

Seven duodenal ulcer patients were treated for 3 months with cimetidine. Before and after treatment endoscopic biopsy specimens were taken for autoradiographic estimation of cell proliferation in the gastric mucosa in the antral and fundic part of the stomach and from the duodenum. In all three areas the estimated labeling index was increased during medication with cimetidine. The increase in epithelial cell renewal may participate in the ulcer healing effect of cimetidine.     

丹参抗大鼠乙醇性胃粘膜损伤的作用及机制

目的 :探讨丹参抗大鼠乙醇性胃粘膜损伤的作用及机制。方法 :采用 10 0 %乙醇复制大鼠乙醇性胃粘膜损伤模型 ,用放射免疫法测定胃粘膜内前列腺素 E2 (PGE2 )、前列腺素 I2 (PGI2 )的含量 ,用浓盐法测定胃粘膜内DNA的含量。结果 :丹参组的胃粘膜损伤指数低于对照组 (P <0 .0 1) ;胃粘膜内 PGE2 、PGI2 含量高于对照组 (均 P<0 .0 1) ;DNA含量高于对照组 (P <0 .0 1)。结论 :丹参促进胃粘膜细胞 DNA的合成和胃粘膜细胞的增殖 ,使PGE2 、PGI2 含量增加 ,进而促进了胃粘液的分泌。     

Paradoxical Role of Helicobacter pylori Infection: Protective Effect Against Ethanol-Induced Gastric Mucosal Injury in Mongolian Gerbils

We investigated the effect of ethanol (a representative necrotizing agent) on gastritis induced by Helicobacter pylori infection in Mongolian gerbils. Seventy-eight gerbils were used. Four and 12 weeks after H. pylori inoculation, 30% ethanol was administered into the stomach. The stomachs were removed after 30 min, the intramucosal prostaglandin (PG) E₂ concentration was measured, and histopathology was recorded. H. pylori infection caused chronic active gastritis, gastric erosion, hypersecretion of mucin from gland mucus cells, and a rise in the activity of intramucosal PGE₂. After ethanol administration, gastric erosion was significantly less in animals infected with H. pylori than in uninfected animals. In conclusion, in the early stage of H. pylori infection, accentuation of intramucosal PGE₂ and hypersecretion of mucin from gland mucus cells have a protective effect against gastric mucosal injury induced by necrotizing agents.     

Effects of Celecoxib on Acid-Challenged Gastric Mucosa of Rats: Comparison with Metamizol and Piroxicam

Selective COX-2 inhibitors have been shown to produce fewer gastrointestinal adverse reactions than classical NSAIDs. Nevertheless, these new agents may worsen and delay the healing of experimentally induced gastric ulcers in animals. In this study, we compared the effects of a selective COX-2 inhibitor (celecoxib), a preferential COX-1 inhibitor (piroxicam), and a nonnarcotic analgesic (metamizol) on normal gastric mucosa of rats and, on the other hand, in a setting of preexisting acute gastric lesions induced by 0.6 N hydrochloric acid. Under normal conditions, only piroxicam produced appreciable gastric lesions. However, after acid challenge the three assayed drugs induced significant macroscopic and microscopic damage. Myeloperoxidase activity as an index of neutrophil infiltration was elevated with celecoxib and piroxicam on normal gastric mucosa. On inflamed mucosa, celecoxib augmented enzymatic activity at the lower dose, which was parallelled by an increase in the interleukin 1beta level. Acid instillaton produced a significant rise in PGE2 content at 7 hr. Drug treatment after acid challenge decreased prostaglandin values in all cases, although to a lesser extent than after single drug dose administration. COX-2 mRNA expression was visible 1 hr after acid application, whereas COX-2 protein could only be detected at 7 hr. Piroxicam increased both expression levels. All NSAIDs enhanced transforming growth factor alpha and epidermal growth factor receptor immunoreactivity around the acid-induced lesions. It is concluded that selective COX-2 inhibitors, like conventional NSAIDs, impair the healing of gastric damage, and therefore special attention should be paid in patients with gastric pathologies.     

Protein Kinase C Involvement in Deoxycholate-Induced Apoptosis in Human Gastric Cells

Bile acids, such as deoxycholic acid (DC), are known to mediate some of their actions by differentially activating various protein kinase C (PKC) isoforms. This study confirms that DC induces apoptosis in gastric epithelial cells through PARP and caspase cascade activation, and examined the role of PKC in DC-induced apoptosis. We found increased activation of PKC in membrane fractions in response to DC that was concentration and time related. The PKC (β_I) isoform expression increased with translocation into the cell membrane fraction after DC (300 μ M) stimulation. In contrast, PKCε expression markedly decreased in response to DC treatment in a time- and concentration-dependent manner. In addition, this process was regulated by caspases, since the pan-caspase inhibitor z-VAD-fmk and caspase-3-, -6-, and -9–specific inhibitors prevented PKC (β_I) and (ε ɛ processing induced by DC. Treatment with the caspase-8–specific inhibitor, however, did not affect expression of either PKC isoform. No significant differences in the apoptotic response were observed when PKC (ɛ) overexpressed cells were exposed to DC in the presence of calcium-dependent conventional PKC inhibitors (Gö 6850 or Gö 6976). Our findings demonstrate that PKC is activated in gastric epithelial cells treated with DC with the PKC (β_I) and PKC (ɛ) isoforms being particularly involved in this process. The processing of PKC (β_I and ε) was shown to be closely regulated by caspases; however, modulations in PKC isoform concentrations by themselves have no effect on the apoptotic death of gastric mucosal cells induced by DC.     

Gastric Epithelial Cell Proliferation in Patients with Liver Cirrhosis

An increased risk for gastric cancer in patients with liver cirrhosis has recently been reported. This study was performed in order to determine gastric epithelial cell proliferation in cirrhotic patients and to evaluate the role of congestive gastropathy (CG) and Helicobacter pylori infection in this process. Thirty-six cirrhotic patients and 18 controls were enrolled in the study. All patients underwent endoscopy and three biopsies were performed in the antrum and three in the gastric body. The presence of H. pylori infection was assessed by a rapid urease test and histology. The antral biopsies were used for gastric cell proliferation assessment by an immunohistochemical analysis (Ki-67). There was no significant difference in epithelial cell proliferation between cirrhotics and controls. Gastric proliferation values were higher in patients with H. pylori infection compared with uninfected patients, both in cirrhotic (P = 0.003) and in control groups (P = 0.06). Among the cirrhotic group, we found a progressive increase in gastric cell proliferation values related to the degree of CG, the highest values being observed in cirrhotic patients with severe CG. Moreover, cirrhotics with both severe CG and H. pylori infection had the highest proliferation values when compared with all other subgroups. In conclusion, this study found that: (1) CG significantly affects epithelial cell proliferation in gastric mucosa in cirrhotic patients, (2) H. pylori infection plays a similar role in gastric cell proliferation in both cirrhotic and non-cirrhotic patients, and (3) CG and H. pylori could act synergistically in this process.     

Stereologic Investigations of Human Gastric Mucosa: II. Oxyntic Mucosa from Patients with Zollinger-Ellison Syndrome

Biopsy specimens from the oxyntic mucosa were obtained on 210 occasions from 76 patients with the Zollinger-Ellison syndrome (ZES) before and during omeprazole treatment. One-micrometer sections were examined by light microscopy, and in 5% linear hyperplasia of endocrine cells was observed. Morphometry was carried out in 91 of the specimens and showed a significant increase of the mean endocrine cell density in comparison with both young, healthy subjects and patients suffering from active peptic ulcer disease (PUD). No metaplasia, dysplasia, or neoplasia was detected in patients with ZES, and the mean mucosal thickness and parietal cell density remained normal. The parietal cells often displayed endosome-like structures, and occasionally there were Ungulate cytoplasmic projections into the gland lumen. Electron microscopic morphometry was carried out in specimens from nine patients with ZES and did not show any significant differences in the parietal cells in comparison with healthy subjects.     

幽门螺杆菌培养上清液诱发小鼠胃粘膜组织损伤的研究

通过动物实验证实幽门螺杆菌(Hp)的细胞毒素对小鼠胃粘膜具有损害作用。用不同剂量的产毒 Hp 菌株(NCTC11637)的培养上清液灌服 BALB/C 小鼠,观察胃粘膜普通病理及超微结构的改变,并与用非产毒 Hp 菌株(来自于临床分离株)的培养上清液及生理盐水灌服过的鼠胃粘膜进行比较。结果表明:Hp 的细胞毒素可以对小鼠胃粘膜产生明显的损害,但并不能引起明显的炎症反应,特别是多形核细胞的浸润。而对照组(用生理盐水)以及无毒素组对小鼠胃粘膜则无明显损害。提示细胞毒素在导致胃部疾病方面起重要作用。     

Pathological Changes in the Formation of Helicobacter pylori-Induced Gastric Lesions in Mongolian Gerbils

We examined pathological changes in theformation of Helicobacter pylori-induced gastric lesionsin Mongorian gerbils. H. pylori (NCTC11637) was orallyadministered once to the animals and was detected in the gastric mucosa of all gerbils given thebacteria. The number of viable H. pylori increasedduring the initial two weeks and thereafter reached aplateau level. The initial pathological changes were found at one week, ie, edema/congestion and awhite viscous substance only in the antrum. At twoweeks, superficial damage appeared in the antrum,although inflammatory cell infiltration had notoccurred. Gastritis with lymphoid follicles was observedin the antrum and fundus from three weeks. At fourweeks, mucosal lesions were detected as a fewhemorrhagic spots in the fundus adjacent to the antrum.In the control animals, however, no pathologicalchanges were observed even at four weeks. In the gastricmucosa infected with H. pylori , myeloperoxidaseactivity was negligible at two weeks, but was extremely elevated at four weeks. Similarly, neutrophilchemotactic activity was only slightly increased at twoweeks, but was markedly elevated at four weeks. Theseresults indicate that H. pylori infection induces initial pathological changes only in theantrum, but mucosal lesions occur in the fundus adjacentto the antrum. Furthermore, it is demonstrated that theinitial superficial damage is generated by factors other than chemokines and neutrophil-associatedfactors, although mucosal inflammation may contribute tothe subsequent formation of lesions andulcers.     

Analysis of Cell Proliferation in Endoscopically Biopsied Specimens of Stomal Mucosa of the Gastric Remnant

Abstract: Carcinoma of the gastric remnant is thought to occur more commonly in the stomal than in the other areas, particularly following Billroth II surgery (BII). An attempt was made to determine the status of cell kinetics in the gastric remnant using endoscopically biopsied specimens, with assay of ornithine decarboxylase (ODC) activity and flow cytometric analysis of the cell cycle parameters. Fifty-two partially gastrectomized patients were studied. Significantly higher ODC activity was recognized in the stomal mucosa than in the greater curvature; this difference was especially pronounced for the group of BII patients. The S-phase cell percentage was also significantly higher in the stomal area than in the greater curvature; this was particularly so in the patients who had undergone BII 5 or more years prior to this study. Our results suggest that activation of cell proliferation is present in the stomal area of the gastric remnant following BII surgery. This activation increased with time following surgery. Activation of cell proliferation occurred in association with histological changes in the stomal glands, and may play a role in carcinogenesis within the gastric remnant.     

胃腺癌者Hp感染胃粘膜的电镜观察

目的：观察Ｈｐ感染胃腺癌者胃粘膜的超微结构形态特点。方法：应用透射电镜技术，对１１例经病理证实为胃腺癌者Ｈｐ感染的粘膜进行观察。结果：可见到Ｈｐ集聚于粘膜上皮绒毛陷窝，位于或接近上皮细胞连接处；主要位于粘液层下，以纤维丝样结构与上皮相连；胃粘膜表面的微绒毛减少或消失，细胞膜肿胀，不规则地形成凹陷或突起；粘液细胞发生粘液排泌、死亡，并观察细胞增殖分裂象。结论：说明Ｈｐ感染的长期存在对细胞的恶性增生有一定影响     

Apoptosis, proliferation and p53 gene expression of H. pylori associated gastric epithelial lesions

AIM:To study the relationship between Helicobacter pylori(H.pylori)and gaatric carcinoma and its possiblepathogenesis by H.pylori.METHODS:DNEL technique and immunohistochemicaltechnique were used to study the state of apoptosis,proliferation and p53 gone expression.A total of 100 gastricmucosal biopsy specimens,including 20 normal mucosa,30H.pylori-negative and 30 H.pylori-positive gastricprecancerous lesions along with 20 gastric carcinomas werestudied.RESULTS:There were several apoptotic cells in thesuperficial epithelium and a few proliferative cells within theneck of gestric glands,and no p53 protein expression innormal mucosa.In gestric carcinoma,there ware fewapoptotic cells,while there were a large number ofproliferative cells,and expression of p53 proteinsignificantly was increased.In the phase of metaplasia,theapoptotic index(Al,4.36%±1.95%),proliferative index(Pl,19.11%±6.79%)and positivity of p53 expression(46.7%)in H.pylori-positive group ware higher than thosein normal mucosa(P<0.01).Al in H.pylori-positive groupwas higher than that in H.pylori-negative group(3.81%±1.76%),Pl in H.pylori-positive group was higher than thatin H.pylori-negative group(12.23%±5.63%,P<0.01).Inthe phase of dysplasia,Al(2.31%±1.10%) in H.pylori-positive group was lower(3.05%±1.29%)than that in H.pylori-negative group,but Pl(33.89%±11.65%)wassignificantly higher(22.09±8018%,P<0.01).In phases ofmetaplasia,dysplasia and gastric cancer in the H.pylori-positive group,Als had an evidently greduall decreasingtrend(P<0.01),while Pls had an evidently gradualincreasing trend(P<0.05 or P<0.01),and there was alsoa trend of gradual increase in the expression of p53 gone.CONCLUSION:In the course of the formation of gastriccarcinoma,proliferation of gastric mucosa can be greatlyIncreased by H.pylori,and H.pylori can induce apoptosisin the phase of metaplasia,but in the phase of dysplesia H.pylorl can inhibit cellular apptosis.And H.pylori infectioncan strengthen the expression of mutated p53 gene.     

Epithelial Apoptosis and Loss in Airways of Children with Asthma

Objective. To examine loss and apoptosis of bronchial epithelial cells in children with asthma. Methods. We examined endobronchial biopsies from 13 asthmatic children and 11 non-asthmatic control subjects with other respiratory diseases. Postmortem samples were obtained from six children who died from non-respiratory diseases. We examined bronchial epithelial shedding by morphology; expression of caspase-3 and terminal deoxynucleotidyl-mediated dUTP nick end labeling (TUNEL) were used to study bronchial epithelial apoptosis. Results. We found epithelial loss to be increased in asthmatic children compared with non-asthmatic control subjects (p = .001) and postmortem children (p = .001). Caspase-3⁺ epithelial cells were significantly greater in children with asthma compared with both non-asthmatic control subjects (p = .001) and the postmortem group (p = .002); TUNEL⁺ epithelial cells were also increased in columnar cells in the asthmatic children compared with the non-asthmatic control subjects (p = .002) and the postmortem group (p = .001). Eosinophilia was absent in 11 of 13 asthmatic children, although they tended to have submucosal lymphocyte infiltration. Smooth muscle and mucus gland hyperplasia were seen in some asthmatic children whose biopsy specimens included these structures. Basement membranes of childhood asthmatics were thicker than in non-asthmatic controls (p = .002) and postmortem subjects (p = .001). Conclusion. Generally, apoptosis and loss of bronchial epithelial cells were increased in childhood asthma; increased apoptosis might be related to epithelial loss.     

Healing of Helicobacter pylori -Induced Gastric Ulcers in Mongolian Gerbils (Combined Treatment with Omeprazole and Clarithromycin)

Helicobacter pylori can colonize the stomachs ofMongolian gerbils and subsequently induce penetratingulcers five months later. Using this gerbil model, theeffects of both combined treatment with omeprazole and clarithromycin, as well as treatment witheach drug separately, on the healing of H.pylori-induced gastric ulcers, and the effects of thecessation of the drug treatment on healed ulcers wereexamined. Beginning five months after H. pylori(NCTC11637) inoculation, omeprazole (four weeks),clarithromycin (two weeks), their combination, or thevehicle was orally administered once daily. These drugs,in combination or separately, markedly enhancedulcer healing and lowered the increased myeloperoxidase(MPO) activity. While omeprazole had no effect on viableH. pylori, clarithromycin and the drug combination significantly reduced viable H. pylori. Thedegree of bacterial eradication was much higher in thecase of the drug combination compared to clarithromycinalone. Four months after cessation of the treatment, visible ulcers, hypertrophic gastritis andincreased MPO activity were found in the control animals(all H. pylori-positive). Nonetheless, only one of theeight gerbils subjected to the drug combination developed a small ulcer, although nohypertrophic gastritis was exhibited. It is concludedthat: (1) the gerbil model of H. pylori infection isuseful for the study of ulcer healing; (2) combinedtreatment with omeprazole and clarithromycin enhances theulcer healing in infected gerbils; and (3) healed ulcersdo not relapse, despite cessation of the drugtreatment.