Does Long-Term Use of Silver Nanoparticles Have Persistent Inhibitory Effect on H. pylori Based on Mongolian Gerbil’s Model?

It is urgent to find alternative agents due to increasing failure rate of Helicobacter pylori (H. pylori) eradication. The study surveyed the long-term effect of silver nanoparticles (AgNP) on H. pylori based on Mongolian gerbil's model

Effects of apigenin on Helicobacter pylori-induced gastritis and gastric cancer in Mongolian gerbils

世界人口中近50 % 人口感染胃幽門螺旋桿菌，在台灣，幽門螺旋桿菌的感染率約為54 %。幽門螺旋桿菌與胃腸道疾病有密切關係，如胃炎，消化道潰瘍及胃癌等疾病。1994年世界衛生組織 (World Health Organization, WHO) 將幽門螺旋桿菌歸類為第一級致癌因子。芹菜素為類黃酮化合物，富含於天然蔬果中，具有抗發炎及抗癌等生理活性。體外試驗中，芹菜素具有顯著地抑制胃腺癌細胞的生長，但卻無任何文獻關於芹菜素對幽門螺旋桿菌引起之胃炎及胃癌影響之體內研究。因此，本研究探討芹菜素對幽門螺旋桿菌誘發蒙古沙鼠胃炎及胃癌的影響。本研究將蒙古沙鼠接種高度致癌活性之幽門螺旋桿菌菌株 (H. pylori KMUH-G926) 進行第一部分芹菜素對於幽門螺旋桿菌誘發蒙古沙鼠胃炎之影響 (滿32週) 及第二部分芹菜素對幽門螺旋桿菌誘發蒙古沙鼠胃癌之影響 (滿52週)，分別研究體重變化、相對器官重量變化並將胃組織進行免疫組織染色以光學顯微鏡進行胃黏膜上皮細胞菌體密度、胃腺體嗜中性白血球浸潤、胃腺體單核白血球浸潤、胃腺體萎縮等項目進行組織病理變化及雪梨分類級分。管餵不同劑量芹菜素 (0-60 mg/kgbw/day) 於各組中，期望芹菜素對幽門螺旋桿菌誘發蒙古沙鼠胃炎及胃癌具有影響作用。結果顯示，第一部分芹菜素對於幽門螺旋桿菌誘發蒙古沙鼠胃炎之各組間體重變化及相對器官重量變化均無顯著性差異，組織病理變化及雪梨分類級分項目以芹菜素 60 mg/kgbw/day處理劑量具有顯著減緩作用分別為芹菜素 0 mg/kgbw/day處理劑量之胃黏膜上皮細胞菌體密度74%、胃腺體嗜中性白血球浸潤66%、胃腺體單核白血球浸潤62%及胃腺體萎縮40%。第二部分芹菜素對於幽門螺旋桿菌誘發蒙古沙鼠胃癌之各組間體重變化及相對器官重量變化均無顯著性差異，組織病理變化及雪梨分類級分項目以芹菜素 60 mg/kgbw/day處理劑量具有顯著減緩作用分別為芹菜素 0 mg/kgbw/day處理劑量之胃黏膜上皮細胞菌體密度65%、胃腺體嗜中性白血球浸潤61%、胃腺體單核白血球浸潤74% 及胃腺體萎縮58%。因此，芹菜素 60 mg/kgbw/day處理劑量對於幽門螺旋桿菌誘發蒙古沙鼠之胃炎及胃癌能有效減緩，其具有高度開發成為治療由幽門螺旋桿菌誘發之胃炎及胃癌藥物之潛力。Nearly 50% of the world''s population is infected by Helicobacter pylori (H. pylori).The overall prevalence rate is 54% in Taiwan. H. pylori infection is known to be associated with the development of gastritis, peptic ulcer and gastric cancer and other diseases. In 1994, H. pylori has been classified by the World Health Organization as a type I carcinogen. Apigenin is a naturally occurring plant flavones and abundantly present in common fruits and vegetables. It is recognized as a bioactive flavonoid and possesses anti-inflammatory and anti-cancer properties. In vitro, apigenin induces apoptosis in human cancer cells. However, In vivo, the effect of ingested apigenin on inflammation and gastric cancer induced by H. pylori remains unkown. Therefore, in this study, we investigated effect of apigenin on inflammation and gastric cancer in Mongolian gerbils model. Mongolian gerbils were orally inoculated with the highly carcinogenic KMUH-G926 strain of H. pylori in the stomach. H. pylori-induced inflammation and gastric cancer in Monoglian gerbils were observed after 32 and 52 weeks (Part I and Part II), respectively. We recorded changes of body weight and relative organ weight, H. pylori densities of gastric mucosa epithelial cells, neutrophil infiltration of gastric glands, monocyte infiltration of gastric glands and atrophy of gastric glands on histopathological changes and Sydney scores in Part I and Part II. Various doses (0-60 mg/kgbw/day) of apigenin were administered in each group. We expect that apigenin can significantly decrease H. pylori-induced inflammatory and gastric cancer incidence in Monoglian gerbils. In the results, there were no significant differences in the body weight and relative organ weight. The treatment dose of 60 mg/kgbw/day of apigenin significantly decreased the histopathological changes and Sydney scores recorded using the induced by H. pylori. After 60 mg/kgbw/day of apigenin treatment, H. pylori densities reduced to compared with the apigenin treatment (0 mg/kgbw/day) was 74% for 32 weeks (Part I)；neutrophil infiltration and moncyte infiltration of gastric glands was 66% and 62%, respectively and gastric glands atrophy was 40%. However, In Part II, there were no significant differences in the body weight and relative organ weight. The treatment dose of 60 mg/kgbw/day of apigenin significantly decreased the histopathological changes and Sydney recorded using the induced by H. pylori. After 60 mg/kgbw/day of apigenin treatment, H. pylori densities reduced to compared with the apigenin treatment (0 mg/kgbw/day) was 65% for 52 weeks (Part II)；neutrophil and monocyte infiltration was 61% and 74%, respectively and gastric glands was 58%. In conclusion, 60 mg/kgbw/day of apigenin treatment significantly decreased the histopathological changes and Sydney scores on the H. pylori-induced inflammation and gastric cancer in Mongolian gerbils. The results suggest that apigenin may has the high potential applications in the drug discovery on the H. pylori-induced inflammation and gastric cancer in Mongolian gerbils therapy.中文摘要………………………………………………………………i 英文摘要………………………………………………………………ii 附表次/表次…………………………………………………………viii 附圖次/圖………………………………………………………………ix 壹、前言………………………………………………………………1 貳、文獻整理…………………………………………………………2 一、幽門螺旋桿菌……………………………………………………2 (一) 幽門螺旋桿菌的發現……………………………………………2 (二) 幽門螺旋桿菌的感染途徑………………………………………2 1.人與人的感染………………………………………………………2 2.口與口的感染………………………………………………………2 3.糞與口的感染………………………………………………………2 4.水源的感染…………………………………………………………3 5.醫療性感染…………………………………………………………3 (三) 幽門螺旋桿菌的盛行率…………………………………………3 (四) 幽門螺旋桿菌的檢測……………………………………………3 1. 須做胃鏡檢查如下…………………………………………………3 (1) 組織學檢查………………………………………………………4 (2) 細菌培養…………………………………………………………4 (3) 聚合酶連鎖反應 (polymerase Chain Reaction) …………4 (4) 快速尿素酶試驗 (rapid urease test) ……………………4 2. 不需做胃鏡檢查如下……………………………………………4 (1) 碳13尿素呼氣法 (urea breath test﹐UBT) ………………4 (2) 血清學檢查 (serology test) ………………………………5 (3) 糞便抗原檢查……………………………………………………5 (五) 幽門螺旋桿菌的致病因子……………………………………5 1.鞭毛 (flagella) …………………………………………………5 2.尿素酶 (urease) …………………………………………………5 3.黏附素 (adhesin) …………………………………………………6 4.細胞毒素相關蛋白 [cytotoxin-associated gene A (cagA) 和 cag pathogenicity island (cag PAI)] …………………………6 5.空泡毒素 (vacuolating cytotoxin A﹐vac A) ………………7 6.嗜中性白血球活化蛋白 (Hp-NAP) ………………………………7 (六) 幽門螺旋桿菌與胃腸道相關疾病………………………………7 1.感染幽門螺旋桿菌時間長短與胃腸道疾病的關係…………………8 2.胃炎 (gastritis) …………………………………………………8 3.消化性潰瘍 (peptic ulcer) ……………………………………8 4.胃癌 (gastric cancer) …………………………………………9 5.環境危險因子與胃癌關係…………………………………………10 (七) 幽門螺旋桿菌的根除治療………………………………………12 1.第一線治療：標準三合療法………………………………………12 2.第二線治療…………………………………………………………12 3.第三線治療…………………………………………………………13 二、幽門螺旋桿菌動物試驗…………………………………………13 (一) 蒙古沙鼠感染幽門螺旋桿菌之胃炎模式……………………13 (二) 蒙古沙鼠感染幽門螺旋桿菌之胃癌模式……………………13 (三) 蒙古沙鼠感染幽門螺旋桿菌與細胞激素關係………………14 三、胃部病理診斷學………………………………………………15 (一) 胃部結構………………………………………………………15 (二) 雪梨分類法 (Sydney system) 之背景………………………15 (三) 雪梨分類法 (Sydney system) 診斷準則……………………16 1. 幽門螺旋桿菌感染密度…………………………………………16 2. 嗜中性白血球浸潤程度…………………………………………16 3. 單核白血球浸潤程度……………………………………………17 4. 萎縮性…………………………………………………………17 5. 腸化生……………………………………………………………17 (四) 病理診斷染色-蘇木紫-伊紅染色…………………………18 四、芹菜素…………………………………………………………19 (一) 天然類黃酮化合物來源及結構特性…………………………19 (二) 芹菜素化學結構及特性………………………………………20 (三) 芹菜素來源……………………………………………………21 (四) 生理活性及化學預防…………………………………………22 參、研究目的…………………………………………………………24 肆、研究架構…………………………………………………………25 伍、材料與方法………………………………………………………26 一、實驗材料…………………………………………………………26 (一) 蒙古沙鼠及飼養環境…………………………………………26 (二) 幽門螺旋桿菌 (Helicobacter pylori) ……………………26 二、實驗方法…………………………………………………………26 (一) 蒙古沙鼠試驗處理……………………………………………26 1. 芹菜素對幽門螺旋桿菌誘發蒙古沙鼠胃炎影響之實驗設計……26 2. 芹菜素對幽門螺旋桿菌誘發蒙古沙鼠胃癌影響之實驗設計……28 (二) 幽門螺旋桿菌培養………………………………………………29 1. 菌株活化……………………………………………………………29 2. 菌株保存…………………………………………………………29 (三) 蒙古沙鼠體重及器官重量變化………………………………30 (四) 幽門螺旋桿菌血清測定………………………………………30 (五) 組織處理及免疫染色…………………………………………30 1. 胃組織處理………………………………………………………30 2. 胃組織脫水………………………………………………………30 3. 胃組織包埋………………………………………………………31 4. 蠟塊組織切片……………………………………………………31 5. 蘇木紫伊紅染色 [hematoxylin & eosin (HE) staining] …31 (六) 組織病理變化觀察……………………………………………31 1. 幽門螺旋桿菌密度 (H. pylori density) ……………………32 2. 嗜中性白血球浸潤程度 (neutrophil infiltration) ………32 3. 單核白血球浸潤程度 (monocyte infiltration) ……………32 4. 胃萎縮之發生 (atrophy) ………………………………………32 5. 胃腺體腸化生 (intestinal metaplasia﹐IM) ………………32 (七) 組織病理胃癌判定(gastric cancer﹐Gca)…………………33 (八) 統計方法…………………………………………………………33 陸、結果與討論………………………………………………………34 第一部份 芹菜素對幽門螺旋桿菌誘發蒙古沙鼠胃炎之影響……34 (一) 幽門螺旋桿菌感染率…………………………………………34 (二) 胃腺體腸化生比率……………………………………………35 (三) 體重變化………………………………………………………35 (四) 相對器官重量變化……………………………………………35 (五) 菌體密度………………………………………………………36 (六) 嗜中性白血球浸潤程度………………………………………37 (七) 單核白血球浸潤程度…………………………………………37 (八) 胃腺體萎縮………………………………………………………………………37 第二部份 芹菜素對幽門螺旋桿菌誘發蒙古沙鼠胃癌之影響……38 (一) 幽門螺旋桿菌感染率…………………………………………38 (二) 胃腺體腸化生比率……………………………………………39 (三) 蒙古沙鼠胃組織模式…………………………………………39 (四) 體重變化………………………………………………………41 (五) 相對器官重量變化……………………………………………41 (六) 菌體密度………………………………………………………42 (七) 嗜中性白血球浸潤程度………………………………………43 (八) 單核白血球浸潤程度…………………………………………43 (九) 胃腺體萎縮……………………………………………………44 柒、討論………………………………………………………………66 捌、參考文獻…………………………………………………………6

Apigenin has anti-atrophic gastritis and anti-gastric cancer progression effects in Helicobacter pylori-infected Mongolian gerbils

Apigenin, one of the most common flavonoids, is abundant in celery, parsley, chamomile, passionflower, and other vegetables and fruits. Celery is recognized as a medicinal vegetable in Oriental countries to traditionally treat inflammation, swelling, blood pressure, serum lipid, and toothache. In this study, we investigated apigenin treatment effects on Helicobacter pylori-induced atrophic gastritis and gastric cancer progression in Mongolian gerbils

Clinical Study Comparison between Single-Dose Esomeprazole-and Pantoprazole-Based Triple Therapy on the Effectiveness for Helicobacter pylori Eradication in Taiwanese Population

Background and Study Aims. To compare the effectiveness of two regimens, single-dose esomeprazole-and pantoprazole-based triple therapy, for Helicobacter pylori (H. pylori) eradication. Patients and Methods. A total of 453 patients were enrolled for H. pylori eradication. They were randomly assigned to either EAC group (Esomeprazole 40 mg once daily, Amoxicillin 1 g twice daily, Clarithromycin 500 mg twice daily for 7 days) or PAC group (Pantoprazole 40 mg twice daily, Amoxicillin 1 g twice daily, Clarithromycin 500 mg twice daily for 7 days). Follow-up endoscopy or urea breath test was scheduled 12-16 weeks after the eradication to evaluate the therapeutic response. Results. Higher eradication rate in EAC group than PAC group was shown by intention-to-treat analysis (EAC 72% versus PAC 55%, P < 0.05) and per-protocol analysis (EAC 91% versus PAC 72%, P < 0.05). The incidence of adverse effects (EAC 19% versus PAC 17%, P = 0.712) and the compliance (EAC 87% versus PAC 91%, P = 0.083) were comparable between these 2 groups. Conclusions. Single-dose esomeprazole-based triple therapy is effective for H. pylori eradication

Comparison between Single-Dose Esomeprazole- and Pantoprazole-Based Triple Therapy on the Effectiveness for Helicobacter pylori Eradication in Taiwanese Population

Background and Study Aims. To compare the effectiveness of two regimens, single-dose esomeprazole- and pantoprazole-based triple therapy, for Helicobacter pylori (H. pylori) eradication. Patients and Methods. A total of 453 patients were enrolled for H. pylori eradication. They were randomly assigned to either EAC group (Esomeprazole 40 mg once daily, Amoxicillin 1 g twice daily, Clarithromycin 500 mg twice daily for 7 days) or PAC group (Pantoprazole 40 mg twice daily, Amoxicillin 1 g twice daily, Clarithromycin 500 mg twice daily for 7 days). Follow-up endoscopy or urea breath test was scheduled 12–16 weeks after the eradication to evaluate the therapeutic response. Results. Higher eradication rate in EAC group than PAC group was shown by intention-to-treat analysis (EAC 72% versus PAC 55%, P<0.05) and per-protocol analysis (EAC 91% versus PAC 72%, P<0.05). The incidence of adverse effects (EAC 19% versus PAC 17%, P=0.712) and the compliance (EAC 87% versus PAC 91%, P=0.083) were comparable between these 2 groups. Conclusions. Single-dose esomeprazole-based triple therapy is effective for H. pylori eradication

Long-Term Use of Probiotic-Containing Yogurts Is a Safe Way to Prevent Helicobacter pylori: Based on a Mongolian Gerbil's Model

Background. The suppression of Helicobacter pylori (H. pylori) decreases H. pylori-related diseases. The probiotics have an inhibitory effect on H. pylori. Aim. We investigated the effects of long-term use of yogurt on H. pylori based on Mongolian gerbils’ model. Materials and Methods. Yogurt (containing a supplement of Lactobacillus acidophilus, Bifidobacterium lactis, etc.) was used. Forty-six gerbils were divided into five groups. All groups were inoculated with H. pylori for 5 to 8 weeks. The yogurt was given as follows: Group (Gr.) A: from 1st to 4th week; Gr. B from 5th to 8th week; Gr. C: from 17th week to sacrifice; Gr. D: from 5th week to sacrifice. Gerbils were sacrificed on the 52nd week. Histology was evaluated according to the Sydney system. Results. The positive rates of H. pylori were 60% (Gr. A), 75% (Gr. B), 67% (Gr. C), 44% (Gr. D), and 100% (Gr. E). Gr. D showed lower inflammatory score. Only Gr. E (60%) had intestinal metaplasia. Gr. D showed higher IL-10 and lower TNF-α expression than Gr. E. Conclusion. Long-term intake of yogurt could decrease H. pylori infection. The long-term use of yogurt would be an alternative strategy to manage H. pylori infection