分泌胃酸的质子泵克隆化

日本大阪大学二井将光教授和前田正知副教授等研究组克隆了分泌胃酸质子泵、(H~++K~+)ATP酶的人染色体基因,在世界上首次在分子水平上弄清了胃酸质子泵的状态。这项研究将解开主要因胃酸分泌过多引起的胃溃疡之谜。目前,这种酶是正在日本进行临床开发的抗溃疡剂即质子泵抑制剂的目标酶。在上述研究成果的基础上正在研究其作用机制。因为是膜蛋白,故结晶困难,但今后立体结构研究清楚后,也许就能从理论上设计出质子泵抑制剂。     

纳米TiO2、ZnO、SiO2对剑尾鱼肝组织中Na+/K+-ATP酶活性的影响

采用浸浴法研究了氧化纳米颗粒TiO2、ZnO、SiO2对剑尾鱼(Xiphophorus helleri)肝中Na+/K+-ATP酶活性的影响。结果表明:纳米TiO2处理组,高浓度(5 mg/L、10 mg/L)组表现为抑制作用,其中5 mg/L处理组Na+/K+-ATP酶的活性与对照组无显著差异(p>0.05),10 mg/L处理组中Na+/K+-ATP酶的活性显著低于对照组中酶的活性(p<0.05)。低浓度组(0.1 mg/L、1 mg/L)则表现为先诱导后抑制,除0.1 mg/L组在暴露1 d后与对照组有显著差异外(p<0.05),其余组与对照组均无显著差异(p>0.05)。纳米ZnO、SiO2处理组(0.1 mg/L、10 mg/L)在暴露1 d后,肝中Na+/K+-ATP酶的活性均比对照组高,随着暴露时间增加至20 d,Na+/K+-ATP酶活性下降,且显著低于对照组(p<0.05)。3种纳米颗粒的浓度为0.1 mg/L时,对暴露后1 d剑尾鱼肝中的Na+/K+-ATP酶活性的影响均为诱导作用,诱导大小顺序为ZnO>TiO2>SiO2;随着暴露时间的增加至10 d,纳米TiO2、ZnO、SiO2处理组对Na+/K+-ATP酶活性均表现出抑制作用。     

盐度对日本鳗鲡(Anguilla japonica)渗透压调节的影响

以日本鳗鲡(505.1±35.7)g为实验对象,分别在淡水(盐度0)、盐度10和盐度33条件下处理14 d,在0、1、4、12、24、96 h和14 d时测定其血清渗透压、离子(Na+、K+、Cl-)浓度和鳃Na+/K+-ATP酶活力指标。结果表明:日本鳗鲡血清等渗点为329.1 m Osm·kg-1,其对应盐度为10.48;随着处理时间延长,盐度处理组血清渗透压、Na+和Cl-浓度均呈现先上升后下降的趋势;血清K+浓度受盐度影响较小(P0.05);盐度10处理组鳃Na+/K+-ATP酶活力于12 h达到最小值(5.40±0.72)μmol·mg-1·h-1,至96 h时恢复至淡水组水平(P0.05);而盐度33处理组鳃Na+/K+-ATP酶活力则表现为先快速下降,后快速上升,并于24 h达到最大值(13.05±0.62)μmol·mg-1·h-1,约为淡水组的1.5倍(P0.05)。日本鳗鲡的渗透压调节可初步分为3个阶段:(1)快速升高期,血清渗透压、Na+和Cl-浓度异常升高,鳃Na+/K+-ATP酶活力受到抑制;(2)缓慢升高期,鱼体补偿失水以缓解渗透压升高,血清渗透压、Na+和Cl-浓度表现为缓慢升高,鳃Na+/K+-ATP酶被激活;(3)适应期,鳃Na+/K+-ATP酶活力处于较高水平,血清渗透压、离子浓度基本恢复。     

磷饥饿时番茄幼苗根系质膜H+-ATP酶活性

磷饥饿提高了番茄幼苗质膜H+-ATP酶活性并促进了番茄幼苗根部的H+分泌。动力学分析表明,磷饥饿使番茄幼苗根部质膜H+-ATP酶的Km值明显降低,亦即提高了该酶对其底物的亲和力,但对该酶的Vmax影响不大。另外,磷饥饿并不改变ATP酶的最适pH值(最适pH值为6.5)。钒酸盐显著抑制番茄幼苗根部质膜ATP酶的活性以及H+分泌,也显著抑制番茄幼苗的Pi吸收。与对照相比,上述抑制作用在饥饿处理的植物中表现得更强。以上结果表明,磷饥饿时高亲和性Pi传递系统的诱导很可能包含质膜H+-ATP酶的参与。     

大豆液泡膜H~+-ATPase的纯化和重组

通过不连续蔗糖密度梯度离心得到的液泡膜微囊 ,先由胆酸钠和 OG分步破膜抽提、经阴离子交换柱 ( Q- Sepharose)层析分离 .纯化后的酶含 V型 H+ - ATPase的主要亚基 ,与大豆磷脂重组 ,获得了有较高泵活性的脂酶体 .脂酶体的质子泵活性受 Valinomycin激活 ,说明它是致电性的 ,受NO-3 ,DCCD以及特异性的 V型 ATPase抑制剂 Bafilomycin的抑制 .脂酶体的泵活性不受 F型和P型 ATPase抑制剂抑制 ,表明质子转运是由 V型 H+ - ATPase引起的 .     

探讨奥美拉唑在临床上的应用

奥美拉唑作为一种新型的胃酸分泌抑制剂,是一种新型的抗消化性溃疡药,对大多数现代抗消化性溃疡药相比,具有更强的抗酸效果。他主要的药理是在胃酸的环境中,能够与壁细胞膜的质子泵相结合抑制其活性,从而减少胃酸的分泌。     

钙对盐胁迫下棉苗离子吸收分配的影响

研究了钙对NaCl胁迫下棉花幼苗体内离子分布的影响及其与根系质膜H+-ATP酶、液泡膜H+-ATP酶和H+-PP酶活性的关系。不同器官离子含量和根系横切面X-射线微区分析结果表明,NaCl胁迫下外源钙明显减少棉花幼苗对Na+的吸收及其向茎杆、叶片的运输,增加对K+和Ca2+的吸收及其向茎杆、叶片的运输,增强棉苗体内的盐分区域化分配,提高根冠比和干物质积累,根系电解质渗漏率下降。钙明显提高盐胁迫下幼根细胞质膜H+-ATP酶、液泡膜H+-ATP酶和H+-PP酶的活性,与钙调节棉花对离子的吸收、分配相一致,说明这些酶可以为根细胞中的Na+在液泡中积累以及K+、Ca2+的选择性吸收和运输提供动力。     

甘草黄酮对长期大强度运动小鼠心肌损伤的保护作用

为了探讨甘草黄酮对长期大强度运动小鼠心肌损伤的保护作用机制。本研究选用50只昆明雄性小鼠为研究对象,以每组10只分组,分为安静对照组(A组)、运动训练组(B组)、运动低剂量给药组(C组)、运动中剂量给药组(D组)、以及运动高剂量给药组(E组)。运动低剂量给药组(C组)、运动中剂量给药组(D组)、运动高剂量给药组(E组)训练前分别灌服剂量为5 g/kg/d、10 g/kg/d、15 g/kg/d黄酮溶液;除安静对照组外,运动组小鼠进行为期6周的大强度游泳训练。测定小鼠各组小鼠血清CK-MB、c Tn I,心肌组织中GSH-PX、SOD、CAT、MDA、NOS、NO、Na+/K+-ATP、Ca2+/Mg2+-ATP。结果表明:与安静对照组比较,运动训练组CK-MB、c Tn I、MDA、NO、NOS升高,GSH-PX、SOD、CAT、Na+/K+-ATP、Ca2+/Mg2+-ATP降低。与运动训练组比较,运动低剂量给药组、运动中剂量给药组、运动高剂量给药组CK-MB、c Tn I、MDA、NO、NOS降低,GSH-PX、SOD、CAT、Na+/K+-ATP、Ca2+/Mg2+-ATP升高;与运动低剂量给药组比较,运动高剂量给药组CK-MB、c Tn I、GSH-Px、SOD、Na+/K+-ATP、Ca2+/Mg2+-ATP升高,MDA降低。本研究提示,长期大强度运动导致机体心肌损伤,甘草黄酮可提高机体心肌组织中抗氧酶、ATPase酶的活性,抑制自由基生成、NOS活性,减少NO的生成,维持细胞膜内外Na+、K+、Ca2+、Mg2+的分布平衡,对大强度运动造成的心肌损伤具有保护作用,效果以高剂量黄酮最佳。     

脑出血并上消化道大出血20例诊治体会

目的:探讨脑出血并上消化道出血的治疗。方法:控制血压、血糖、降颅压及预防并发症,另应用止血剂、输血及补充血容量、静脉用H2受体阻滞剂。用生长抑素止血及用质子泵抑制剂制止胃酸分泌。     

燕麦幼苗对盐胁迫的响应及过氧化氢对响应的调节

为了探讨‘定莜6号’燕麦对盐胁迫的生理生化响应及H2O2的调节作用,采用水培方法,研究外源H2O2对盐胁迫下燕麦活性氧代谢、渗透溶质积累和Na+、K+平衡的影响。结果表明:小于100 mmol·L-1Na Cl未对‘定莜6号’幼苗的生长造成明显影响,150 mmol·L-1及以上浓度Na Cl使幼苗干重和叶片K+/Na+显著降低,O2-·产生速率、H2O2、丙二醛(MDA)、可溶性蛋白质和脯氨酸含量及过氧化氢酶(CAT)、质膜H+-ATP酶活性明显提高,但抗坏血酸(ASA)和谷胱甘肽(GSH)含量变化不大;外施5μmol·L-1H2O2可显著缓解150 mmol·L-1Na Cl胁迫对燕麦幼苗生长的抑制作用,使燕麦叶片超氧化物歧化酶(SOD)、抗坏血酸过氧化物酶(APX)和CAT活性及H2O2、GSH含量明显提高,O2-·产生速率和MDA含量显著降低;5μmol·L-1H2O2还提高了150 mmol·L-1Na Cl胁迫下燕麦叶片可溶性蛋白质、可溶性糖、有机酸和脯氨酸含量及质膜H+-ATP酶活性和K+/Na+,降低了游离氨基酸含量;表明外源H2O2可调控燕麦幼苗活性氧代谢和渗透溶质积累,维持K+、Na+平衡,从而增强耐盐性。     

Discovery, synthesis, and biological evaluation of novel pyrrole derivatives as highly selective potassium-competitive acid blockers

To discover a gastric antisecretory agent more potent than existing proton pump inhibitors, novel pyrrole derivatives were synthesized, and their H(+),K(+)-ATPase inhibitory activities and inhibitory action on histamine-stimulated gastric acid secretion in rats were evaluated. Among the compounds synthesized, compound 17a exhibited selective and potent H(+),K(+)-ATPase inhibitory activity through reversible and K(+)-competitive ionic binding; furthermore, compound 17c exhibited potent inhibitory action on histamine-stimulated gastric acid secretion in rats and Heidenhain pouch dogs.     

pH, healing rate, and symptom relief in patients with GERD

Gastroesophageal reflux symptoms are common and occur in all of us from time to time. In others, reflux may be associated with ulcerative esophagitis. The symptoms may be aggravated by large meals, coffee, smoking and position. Physiological and pathological reflux can be separated by the frequency and duration of the exposure of the lower esophagus to acid. Pathological reflux results in symptoms and also esophagitis and ulceration in some patients. Although gastroesophageal reflux disease (GERD) is considered to result from a disorder of motility in the esophagus, gastric acid and peptic activity are deemed pivotal to the initiation and continuation of the esophageal damage and the development of symptoms. Acid exposure in the esophagus is normally less than 4 percent of the 24 hours with a pH below 4. An increase over 4 percent of the time with a pH less than 4 is considered pathological. Hence, antisecretory drugs have become the principle approach to the treatment of reflux symptoms and esophagitis since they reduce the acidity, of gastric juice and the activity of pepsin. Importantly, they also reduce the volume of gastric juice available for reflux into the esophagus. There is a clear relationship between the degree and duration of acid suppression and the relief of heartburn and healing of esophagitis. Pharmacodynamic studies with different dose regimens of the H2-receptor antagonists and the proton pump inhibitors show a difference in the degree and duration of the antisecretory effect, and this correlates closely with the results of clinical trials with respect to the healing of esophagitis and the relief of symptoms. Proton pump inhibitors achieve healing rates by week four, which are not achieved by H2-receptor antagonists even after 12 weeks of treatment. The advantage of proton pump inhibitors over H2-receptor antagonists is due to the greater degree, longer duration of effect and more complete inhibition of acid secretion that maintains intragastric pH above 4 for a maximal duration. Although there is no significant difference between proton pump inhibitors with respect to healing of esophagitis, symptom relief occurs earlier with lansoprazole than omeprazole, and this is probably due to the greater oral bioavailability and faster onset of action of lansoprazole when compared to omeprazole.     

Omeprazole-like compounds on histamine-stimulated acid and peptic secretions in conscious dog and cat.

A newly synthetized aniline derivative series has been tested on histamine-stimulated gastric acid secretion in conscious cats and dogs and compared to the proton pump inhibitors omeprazole and NC 1300. The compounds markedly and long-lastingly inhibited acid output with a potency lower than omeprazole but very close to the reference compound NC 1300. The results show that the introduction of a nitrogen atom, irrespective of the position in the benzimidazole ring does not seem to influence the gastric antisecretory activity.     

Pharmacotherapy for acid/peptic disorders.

In the 1970s, the identification of the histamine H2-receptor by Black and the subsequent development of histamine H2-receptor antagonists revolutionized our understanding and treatment of acid/peptic disorders. More recently, the identification of hydrogen-potassium-stimulated adenosine triphosphatase (H+/K(+)-ATPase) as the proton pump of the parietal cell and the recognition of the prominent role of Helicobacter pylori in the pathogenesis of duodenal and gastric ulceration have heralded a new revolution in our understanding and treatment of these disorders. Substituted benzimidazole compounds (omeprazole, lansoprazole and pantoprazole) that covalently bind to and inactivate the proton pump allow complete and prolonged inhibition of acid secretion. Not only can peptic ulcers now be healed more rapidly with proton pump inhibitors, but refractory ulcers have all but disappeared. Eradication of H. pylori with antibiotics offers, for the first time, a permanent cure for most duodenal and many gastric ulcers.     

pH,healing rate and symptom relief in acid-related diseases.

Suppression of gastric acid secretion is widely used and logical for the treatment of acid-related diseases. Healing of duodenal ulcer, gastric ulcer and gastroesophageal reflux disease is correlated significantly with the degree and the duration of suppression of intragastric acidity over 24 hours and with the length of the treatment. To date, proton pump inhibitors are the most effective agents among the currently available antisecretory drugs in offering the highest healing rate and fastest resolution of symptoms. Combinations of an antisecretory drug with one or more antimicrobial agents accelerate healing of peptic ulcers.     

Characteristics of the K+-competitive H+,K+-ATPase inhibitor AZD0865 in isolated rat gastric glands

The gastric H+,K+-ATPase of the parietal cell is responsible for acid secretion in the stomach and is the main target in the pharmacological treatment of acid-related diseases. Omeprazole and other benzimidazole drugs, although having delayed efficacy if taken orally, have high success rates in the treatment of peptic ulcer disease. Potassium competitive acid blockers (P-CAB) compete with K+ for binding to the H+,K+-ATPase and thereby they inhibit acid secretion. In this study, the in vitro properties of AZD0865, a reversible H+,K+-ATPase inhibitor of gastric acid secretion, are described. We used a digital-imaging system and the pH sensitive dye BCECF to observe proton efflux from hand-dissected rat gastric glands. Glands were stimulated with histamine (100 microM) and exposed to a bicarbonate- and Na+-free perfusate to induce an acid load. H+,K+-ATPase inhibition was determined by calculating pHi recovery (dpH/dT) in the presence of omeprazole (10-200 microM) or AZD0865 (0.01-100 microM). The efficacies of both drugs were compared. Our data show that acid secretion is inhibited by both the proton pump inhibitor omeprazole and the P-CAB AZD0865. Complete inhibition of acid secretion by AZD0865 had a rapid onset of activation, was reversible, and occurred at a 100-fold lower dose than omeprazole (1 microM AZD0865 vs. 100 microM omeprazole). This study demonstrates that AZD0865 is a potent, fast-acting inhibitor of gastric acid secretion, effective at lower concentrations than drugs of the benzimidazole class. Therefore, these data strongly suggest that AZD0865 has great potential as a fast-acting, low-dose inhibitor of acid secretion.     

Recent advances in proton pump inhibitors and management of acid-peptic disorders

Acid-peptic ulcers and diseases have been increasingly on rise in today's era of globalization, which is characterized by hurry, worry, and curry. This review summarizes various disorders associated with increased gastric acid secretion and various therapeutic strategies to control them. The emphasis has been laid, in particular, on the role of proton pump inhibitors (PPIs) widely used nowadays for the treatment of gastric acid diseases. The medicinal chemistry aspects and mechanism of action of irreversible PPIs and APAs have been discussed at molecular levels. The ongoing research status in this field has also been covered. Further, biological evaluation methods that can be used for screening of PPIs are also discussed in short.     

The biochemical and pharmacological properties of a newly synthesized H+-K+ ATPase inhibitor, 2-dimethylamino-4,5-dihydrothiazolo[4,5:3,4]pyridol-[1,2-a]benzimidazol e

This study was designed to determine the effect of a newly synthesized proton pump inhibitor, 2-dimethylamino-4,5-dihydrothiazolo[4,5:3,4]pyridol[1,2-a]be nzimidazole (YJA20379-2), on gastric H(+)-K(+) ATPase activity, acid secretion, and experimental gastroduodenal lesions or ulcers in rats. YJA20379-2 inhibited in a concentration-dependent manner the proton pump (H(+)-K(+) ATPase) activity in isolated hog gastric mucosal microsomes, therefore, confirming its classification as a proton pump inhibitor. The inhibitory efficacy of YJA20379-2 on the proton pump was about eight times higher than that of omeprazole at pH 7.4. The activity of the inhibited enzyme was not restored by dilution and washout method, so this implied that the inhibition of YJA20379-2 is not reversible. YJA20379-2, given intraduodenally or orally, potently suppressed acid secretion in pylorus-ligated rats, with ED50 values of 3.6 and 7.7 mg.kg(-1), respectively. Pretreatment with YJA20379-2 dose dependently protected the gastric mucosa from damage induced by absolute ethanol, water-immersion stress, indomethacin, and the duodenal mucosa from damage induced by mepirizole in rats, with ED50 values of 11.0, 21.0, 0.5, and 18.7 mg.kg(-1), respectively. Repeated administration of YJA20379-2 also dose dependently accelerated spontaneous healing of acetic acid induced gastric ulcers. These results suggest that YJA20379-2 has potent antisecretory and antiulcer effects, which are exerted by suppression of H(+)-K(+) ATPase activity in gastric parietal cells, such that YJA20379-2 may be useful for the clinical treatment of peptic ulcer diseases.     

Helicobacter pylori modulation of gastric acid

Helicobacter pylori plays major causative roles in peptic ulcer disease and gastric cancer. Elevated acid secretion in patients with duodenal ulcers (DUs) contributes to duodenal injury, and diminished acid secretion in patients with gastric cancer allows carcinogen-producing bacteria to colonize the stomach. Eradication of H. pylori normalizes acid secretion both in hyper-secreting DU patients and hypo-secreting relatives of gastric cancer patients. Therefore, we and others have asked how H. pylori causes these disparate changes in acid secretion. H. pylori gastritis more or less restricted to the gastric antrum in DU patients is associated with increased acid secretion. This is probably because gastritis increases release of the antral acid-stimulating hormone gastrin and diminished mucosal expression of the inhibitory peptide somatostatin. Bacterial products and inflammatory cytokines including TNFalpha may cause these changes in endocrine function. Gastritis involving the gastric corpus tends to diminish acid secretion, probably because bacterial products and cytokines including IL-1 inhibit parietal cells. Pharmacological inhibition of acid secretion increases corpus gastritis in H. pylori-infected subjects, so it is envisaged that gastric hypo-secretion of any cause might become self-perpetuating. H. pylori-associated mucosal atrophy will also contribute to acid hypo-secretion and is more likely in when the diet is high in salt or lacking in antioxidant vitamins. Data on gastric acid secretion in patients with esophagitis are limited but suggest that acid secretion is normal or slightly diminished. Nevertheless, H. pylori infection may be relevant to the management of esophagitis because: (i) H. pylori infection increases the pH-elevating effect of acid inhibiting drugs; (ii) proton pump inhibitors may increase the tendency of H. pylori to cause atrophic gastritis; and (iii) successful eradication of H. pylori is reported to increase the likelihood of esophagitis developing in patients who had DU disease. Points (ii) and (iii) remain controversial and more work is clearly required to elucidate the relationship between H. pylori, acid secretion, gastric mucosa atrophy and esophagitis.     

Current status of acid pump antagonists (reversible PPIs).

The introduction of H2-receptor antagonists in the mid-1970s provided, for the first time, acceptable medical therapy for acid-related diseases. Their short duration of action and single receptor targeting, however, limited satisfactory treatment of patients. Today the control of gastric acid secretion can be effectively achieved by direct inhibition of the H+, K(+)-ATPase. Inhibition of the proton pump suppresses acid secretion independent of the route of stimulation. Two classes of drugs are able to inhibit the proton pump. First, the substituted benzimidazoles (proton pump inhibitors [PPIs]), which, due to their pKa of about 4, accumulate in the acidic secretory canaliculus of the stimulated parietal cell. Following conversion to a cationic sulfenamide, they react with cysteines on the extracytoplasmatic face of the H+, K(+)-ATPase subunit. Second, acid pump antagonists (APAs) acting by K(+)-competitive and reversible binding to the gastric proton pump, which is the final step for activation of acid secretion in the parietal cell. One possible class of APAs are imidazopyridines. BY841 was selected from this class and is chemically a (8-(2-methoxycarbonylamino-6-methyl-phenylmethylamino )-2,3-dimethyl-imidazo [1,2-a]-pyridine). In pharmacological experiments such as pH-metry in the conscious, pentagastrin-stimulated fistula dog, BY841 proved to be superior to both ranitidine and omeprazole by rapidly elevating intragastric pH up to a value of 6. The duration of this pH elevation in the dog was dose-dependent. As was predicted by the above-mentioned dog model, available clinical phase I data confirm dose-dependent pharmacodynamics of BY841 in man. Using both acid output and continuous 24-hr pH measurements, a pronounced antisecretory effect of BY841 has been found. Actually, a single 50 mg oral dose of BY841 immediately elevated intragastric pH to about 6. Higher doses caused a dose-dependent increase in duration of the pH-elevation, without any further increase in maximum pH values. Twice daily administration was more effective than once a day administration of the same daily dose. With both regimens, the duration of the pH-elevating effect of BY841 further increased upon repeated daily administration. This demonstrates lack of tolerance development, the latter being a well-known disadvantage of H2-receptor antagonists. In comparison with the standard dose of omeprazole, BY841 administered at a dose of 50 mg or 100 mg twice daily is markedly more effective on Day one of treatment, and both doses are at least as potent as omeprazole following repeated daily administration.