Establishment of Mongolian gerbil model of gastric cancer induced by Helicobacter pylori infection and its proteomics analysis / 中华病理学杂志

<p><b>OBJECTIVE</b>To establish an animal model of gastric cancer by long-term infection of Helicobacter pylori (H.pylori) and to elucidate the pathogenesis by proteomics analysis.</p><p><b>METHODS</b>Fifty male Mongolian gerbils (4-5 week-old and weighted 60-100 g) were infected with H.pylori and the gastric tissues were obtained after the infection at 3, 6, 12 and 24 months. Histological changes were evaluated by H-E staining of the gastric tissue sections. Detection of H.pylori was performed by in-vitro culture of fresh gastric tissue samples, PCR amplification of H.pylori 16s rRNA and localization by silver staining. In addition, proteins extracted from gastric tissue samples were subjected to two-dimensional electrophoresis (2-DE) at various infection time points. Protein spots with increased quantity over the course of H.pylori infection were selected and analyzed by LC-MS/MS. Finally, differentially expressed proteins between human gastric cancer tissue samples and lymph nodes were analyzed by real-time RT-PCR.</p><p><b>RESULTS</b>Colonization of H.pylori was observed in gastric tissue of gerbils as early as 3 months after H.pylori infection, and persisted till 24 months. Pathological examination of infected animals showed various histological changes including acute gastritis, atrophic gastritis, intestinal metaplasia and gastric carcinoma. Seventy-eight differentially expressed proteins were identified by proteomics analysis, among which 36 proteins were up-regulated and 42 were down-regulated. Analyzed by LC-MS/MS, ten proteins were identified, including lactate dehydrogenase, ATP synthase, fatty acid-binding protein, COX5B, peroxiredoxin-4, peroxide reductase, transgelin, succinyl-CoA ligase, keratin and protein disulfide-isomerase A2, among which transgelin, ATP synthase and lactate dehydrogenase were highly expressed in human gastric carcinoma and lymph nodes.</p><p><b>CONCLUSIONS</b>H.pylori infection induces the expression of transgelin, ATP synthase and lactate dehydrogenase, implying possible roles in the pathogenesis of gastric diseases including cancer.</p>

Altered Regulation of Renal Acid Base Transporters in Response to Ammonium Chloride Loading in Rats

The role of the kidney in combating metabolic acidosis has been a subject of considerable interest for many years. The present study was aimed to determine whether there is an altered regulation of renal acid base transporters in acute and chronic acid loading. Male Sprague-Dawley rats were used. Metabolic acidosis was induced by administration of NH4Cl for 2 days (acute) and for 7days (chronic). The serum and urinary pH and bicarbonate were measured. The protein expression of renal acid base transporters [type 3 Na+/H+ exchanger (NHE3), type 1 Na+/HCO3- cotransporter (NBC1), Na-K+ ATPase, H(+)-ATPase, anion exchanger-1 (AE-1)] was measured by semiquantitative immunoblotting. Serum bicarbonate and pH were decreased in acute acid loading rats compared with controls. Accordingly, urinary pH decreased. The protein expression of NHE3, H(+)-ATPase, AE-1 and NBC1 was not changed. In chronic acid loading rats, serum bicarbonate and pH were not changed, while urinary pH was decreased compared with controls. The protein expression of NHE3, H(+)-ATPase was increased in the renal cortex of chronic acid loading rats. These results suggest that unaltered expression of acid transporters combined with acute acid loading may contribute to the development of acidosis. The subsequent increased expression of NHE3, H(+)-ATPase in the kidney may play a role in promoting acid excretion in the later stage of acid loading, which counteract the development of metabolic acidosis.

Biopsy-Proven Type 1 Renal Tubular Acidosis in a Patient with Metabolic Acidosis

No abstract available.

Genetic studies in a family with distal renal tubular acidosis and sensorineural deafness.

Distal renal tubular acidosis (RTA) with sensorineural deafness is a rare entity, inherited in an autosomal recessive manner. It is caused by mutations in the ATP6V1B1 gene, leading to defective function of H+-ATPase pump in the distal nephron, cochlea and endolymphatic sac. We report two siblings with distal RTA and sensorineural deafness having mutation C>T in the first coding exon of the gene, resulting in a non functional protein. The parents were found to be carriers for the mutation.

Mutation analysis of ATP13A2 gene in Chinese patients with familial autosomal recessive early-onset parkinsonism / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the mutation characteristics of ATP13A2 gene in Chinese patients with familial autosomal recessive early-onset parkinsonism (AREP).</p><p><b>METHODS</b>Mutations of ATP13A2 gene were screened by polymerase chain reaction combined with DNA direct sequencing in patients with familial AREP.</p><p><b>RESULTS</b>No pathogenic mutations in ATP13A2 gene were detected in this group. Six reported polymorphisms were identified. They were IVS6+70A>G, IVS12+66A>G, m.1849C>T, IVS20-56 G>A, m2671C>T and m2824G>A.</p><p><b>CONCLUSION</b>ATP13A2 gene mutations may be rare in Chinese patients with familial autosomal recessive early-onset parkinsonism.</p>

Effects of extremely low frequency magnetic fields on hydrolysis of F0F1-ATPases and their relationship with turnover rates of F1 / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To study the effects of extremely low frequency sinusoidal magnetic fields on hydrolysis of F(0)F(1)-ATPase and its mechanism.</p><p><b>METHODS</b>The F(0)F(1)-ATPases which was localized on the outer surface of chromatophores were prepared from the cells of Rhodospirillum rubrum and were exposed to 0.1 approximately 0.5 mT, 4.7 approximately 96.0 Hz magnetic fields.</p><p><b>RESULTS</b>The hydrolysis activity of F(0)F(1)-ATPase was stimulated by 0.5 mT, 4.7, 12.0, 60.0, 72.0, 84.0 and 96.0 Hz magnetic fields respectively and inhibited by 0.5 mT, 24.0 Hz magnetic field (P < 0.05); 0.3 mT, 4.7, 24.0 and 60.0 Hz magnetic fields also distinctly affected F(0)F(1)-ATPases activity respectively (P < 0.05), whereas 0.1 mT exposure caused no significant changes on that activity. When the hydrolysis activity of the F(0)F(1)-ATPases was inactivated by its inhibitor DCCD, the 0.5 mT, 24.0 Hz magnetic field still inhibited the hydrolysis activity of the F(0)F(1)-ATPase and 0.5 mT, 60.0 Hz magnetic field also had stimulating effects (P < 0.05).</p><p><b>CONCLUSION</b>The effects of magnetic fields on the hydrolysis activity of the F(0)F(1)-ATPases depend on not only magnetic frequency but also magnetic intensity. The threshold of magnetic intensity is between 0.1 mT and 0.3 mT. F(0)F(1)-ATPases, especially F1-portion may be an end-point of magnetic fields.</p>

Cl- and regulation of pH by MDCK-C11 cells

The interaction between H+ extrusion via H+-ATPase and Cl- conductance was studied in the C11 clone of MDCK cells, akin to the intercalated cells of the collecting duct. Cell pH (pHi) was measured by fluorescence microscopy using the fluorescein-derived probe BCECF-AM. Control recovery rate measured after a 20 mM NH4Cl acid pulse was 0.136 ± 0.008 pH units/min (dpHi/dt) in Na+ Ringer and 0.032 ± 0.003 in the absence of Na+ (0 Na+). With 0 Na+ plus the Cl- channel inhibitor NPPB (10 æM), recovery was reduced to 0.014 ± 0.001 dpHi/dt. 8-Br-cAMP, known to activate CFTR Cl- channels, increased dpHi/dt in 0 Na+ to 0.061 ± 0.009 and also in the presence of 46 nM concanamycin and 50 æM Schering 28080. Since it is thought that the Cl- dependence of H+-ATPase might be due to its electrogenic nature and the establishment of a +PD (potential difference) across the cell membrane, the effect of 10 æM valinomycin at high (100 mM) K+ was tested in our cells. In Na+ Ringer, dpHi/dt was increased, but no effect was detected in 0 Na+ Ringer in the presence of NPPB, indicating that in intact C11 cells the effect of blocking Cl- channels on dpHi/dt was not due to an adverse electrical gradient. The effect of 100 æM ATP was studied in 0 Na+ Ringer solution; this treatment caused a significant inhibition of dpHi/dt, reversed by 50 æM Bapta. We have shown that H+-ATPase present in MDCK C11 cells depends on Cl- ions and their channels, being regulated by cAMP and ATP, but not by the electrical gradient established by electrogenic H+ transport.

Identification of Differentially Expressed Genes in Murine Hippocampus by Modulation of Nitric Oxide in Kainic Acid-induced Neurotoxic Animal Model

Kainic acid (KA) causes neurodegeneration, but no consensus has been reached concerning its mechanism. Nitric oxide may be a regulator of the mechanism. We identified differentially expressed genes in the hippocampus of mice treated with kainic acid, together with or without L-NAME, a nonselective nitric oxide synthase inhibitor, using a new differential display PCR method based on annealing control primers. Eight genes were identified, including clathrin light polypeptide, TATA element modulatory factor 1, neurexin III, ND4, ATPase, H+ transporting, V1 subunit E isoform 1, and N-myc downstream regulated gene 2. Although the functions of these genes and their products remain to be determined, their identification provides insight into the molecular mechanism(s) involved in KA-induced neuronal cell death in the hippocampal CA3 area.

Effects of valsartan and captopril on expressions and activities of tissue factor and tissue factor pathway inhibitor / 中华心血管病杂志

<p><b>OBJECTIVE</b>To investigate the effects of ox-LDL, ACEI and ARB on expressions and activities of TF and TFPI in VSMC.</p><p><b>METHODS</b>(1) Rabbit VSMC was cultured by explant-attached method in vitro. (2) The effects of ox-LDL and valsartan on TF and TFPI expressions were analyzed by immunohistochemistry and immunofluorescence. Laser scanning confocal microscopy were applied to analyze the effects of ox-LDL and valsartan on TF expression. The effects of ox-LDL, valsartan and captopril on TF and TFPI antigen expressions were analyzed by ELISA. Chromogenic substrate method was used to determine the effects of ox-LDL, valsartan and captopril on TF activity. The effects of ox-LDL and valsartan on TF mRNA expression were analyzed by RT-PCR.</p><p><b>RESULTS</b>(1) ox-LDL could upregulate TF antigen, activity and TF expression at mRNA level and downregulate TFPI antigen. (2) Valsartan and captopril could reduce TF antigen and activity in VSMC treated by ox-LDL, and valsartan reduce it in a dose-dependent manner. Valsartan could also attenuate TF expression at mRNA level in VSMC treated by ox-LDL. (3) Using ELISA, valsartan and captopril could also enhance TFPI antigen in VSMC treated by ox-LDL.</p><p><b>CONCLUSION</b>Our study showed upregulated TF and downregulated TFPI expression and activity by ox-LDL and these effects could be reversed by ACEI and ARB indicating a new insight on the antiatherosclerotic effects of ACEI and ARB.</p>

Effects of seawater immersion on the functions of mitochondria of myocardium and hepatocyte in hemorrhagic shock rats / 中华创伤杂志(英文版)

<p><b>OBJECTIVE</b>To investigate the effects of seawater immersion on the function of myocardium and hepatocyte mitochondria in experimental hemorrhagic shock rats.</p><p><b>METHODS</b>Twenty-four male Wistar rats were divided into three groups (n=8 in each group): control group, HSL group (hemorrhagic shock group on land) and HSS group (hemorrhagic shock group in seawater). The hemodynamic parameters, activities of H(+)-ATPase (adenosinetriphosphatase), succinate dehydrogenase (SDH) and Ca(2+)-Mg(2+)-ATPase, the calcium contents in myocardium and hepatocyte mitochondria were measured and the changes of proton translocation across the inner mitochondrial membrane were analyzed.</p><p><b>RESULTS</b>The hemodynamic indexes and the activities of H+-ATPase, SDH, Ca(2+)-Mg(2+)-ATPase in HSS group were significantly lower than those in control group and HSL group (P<0.05). In HSS group the calcium levels in tissue and mitochondria of myocardium and hepatocyte were elevated significantly compared with control group and HSL group (P<0.05). There was no significant difference in proton translocation among three groups.</p><p><b>CONCLUSIONS</b>This investigation demonstrates that seawater immersion can aggravate the conditions of hemorrhagic shock rats.</p>

A Randomized, Prospective, Comparative, Multicenter Study of Rabeprazole and Ranitidine in the Treatment of Reflux Esophagitis / 대한소화기학회지

BACKGROUND/AIMS: This study was done to evaluate the efficacy of rabeprazole (proton-pump-inhibitor) and ranitidine (H2-receptor antagonist) in the symptom relief and treatment of erosive esophagitis diagnosed by endoscopy. METHODS: A total of 110 patients with typical gastroesophageal reflux disease (GERD) symptoms were enrolled in this multicenter study. They were randomized into rabeprazole group (53 patients) and ranitidine group (57 patients) respectively. The patients in rabeprazole group were given 10 mg of rabeprazole and ranitidine group received 300 mg of ranitidine before breakfast and dinner for 8 weeks. After the end of treatment, we evaluated the endoscopic healing rate of reflux esophagitis and symptomatic improvement. RESULTS: After 8 weeks of treatment, rabeprazole group showed significantly higher complete endoscopic cure rate than ranitidine group (86.8% [46/53] vs. 57.9% [33/57], p=0.001) and higher symptomatic improvement of heartburn (91.2% [31/34] vs. 76.2% [32/42], p=0.085), especially in the first 7 days (76.7% vs. 45.3%, p=0.008). Also, rabeprazole group showed significantly higher improvement of regurgitation symptom than ranitidine group (100% [35/35] vs. 83% [39/47], p=0.009). Both group showed no differences in the improvement of chest pain and globus sensation. All the adverse events (rabeprazole group 4 events vs. ranitidine group 3 events) were mild and there was no abnormality in laboratory test. CONCLUSIONS: In patients with GERD, rabeprazole 10 mg b.i.d. is superior to ranitidine 300 mg b.i.d. in healing of reflux esophagitis and resolving typical GERD symptoms. Rabeprazole is an effective and well-tolerated drug for GERD treatment.

Efeito de uma semana de esomeprazol associado a claritromicina e amoxicilina para a erradicação do helicobacter pylori e cicatrização da úlcera duodenal estudo aberto, não randomizado, não controlado e multicêntrico / Effect of 1-week treatment with esomeprazole plus clarithromycin and amoxicillin to eradicate Helicobacter pylori, and to heal duodenal ulcer. Open, non-randomized, non-controlled, multicenter study

A úlcera péptica é causada, na sua maioria, por infecção pelo Helicobacter pylori, sendo portanto recomendada sua erradicação. Um dos tratamentos da erradicação mais frequentemente utilizado é o esquema tríplice baseado em inibidor de bomba protônica associado a dois antimicrobianos. O inibidor da bomba de prótons freqüentemente é continuado por mais três semanas com monoterapia, após o esquema de erradicação, para assegurar cicatrização da úlcera e alívio dos sintomas, em pacientes com úlcera duodenal ativa. O esomeprazol é um dos isômeros ópticos dos composto racêmico omeprazol e, assim, tem o mesmo mecanismo de ação, porém com área sob a curva (AUC) maior e menor variabilidade polimórfica em seu metabolismo hepático. Assim, é possível melhor eficácia clínica do esomeprazol, com efeito inibitório mais duradouro e profundo sobre a secreção ácida gástrica durante o período de 24 horas e menor variação interindividual na acidez gástrica. Vários trabalhos publicados mostraram que o tratamento de erradicação do H. Pylori com esquema tríplice baseado em esomeprazol, por uma semana, tem a mesma eficácia na cicatrização da úlcera duodenal que o tratamento tríplice baseado em omeprazol, por uma semana, continuado por três semanas com omeprazol. Este estudo busca confirmar esses dados em centros brasileiros. O objetivo primário deste estudo foi avaliar as taxas de cicatrização em pacientes com úlcera duodenal, após uma semana de tratamento de erradicação do H. pylori, seguida por quatro semanas de observação sem tratamento. Os pacientes receberam tratamento de erradicação com a terapia tríplice esomeprazol 20mg bid + amoxilina 1.000mg bid + claritromicina 500mg bid, administrados por sete dias. Os objetivos segundários foram: avaliar as taxas de erradicação do H. Pylori; determinar a freqÜência e intensidade de sintomas como azia/pirose e dor epigástrica; avaliar a segurança e a tolerabilidade do esomeprazol em combinação com claritomicina e amoxicilina. O estudo foi aberto, não randonizado, não controlado, participando 12 centros no Brasil. No total, 144 pacientes foram analidados quando a eficácia clínica (análise por protocolo - PP). Os pacientes foram acompanhados por cinco semanas com duas visitas clínicas agendadas durante esse período. A dor epigástrica estava presente em 97,2por cento(140/144) dos pacientes no inicio do estudo

Efeito do hormônio tireoidiano sobre a expressão do RNAm da proteína desacopladora de protóns 3 (UCP3) em miocárdio e músculo esquelético de ratos / Effect of thyroid hormone on UCP-3 mRNA expression in rat heart and skeletal muscle

As proteínas desacopladoras de prótons (UCPs) pertencem à família dos transportadores mitocondriais H+/ácidos graxos. A UCP1 é responsável pela termogênese, mas o exato papel fisiológico da UCP2 e UCP3 ainda não está estabelecido. Ratos foram induzidos ao hipertireoidismo, através da administração de LT3, associado ou não à captopril, propranolol ou clenbuterol. O tratamento com LT3 resultou em aumento estatisticamente significativo do mRNA da UCP3 em miocárdio e músculo esquelético e esse efeito não foi alterado por nenhuma das medicações usadas concomitantemente, nem quando o LT3 foi administrado conjuntamente ao clenbuterol. Os efeitos do hormônio tireoideano sobre a expressão da UCP3, não são dependentes da angiotensina II, nem do sistema β-adrenérgico, refletindo uma ação direta do T3 sobre a expressão da UCP3 / Thyroid hormones stimulate UCP3 expression in skeletal muscle. We examined whether thyroid hormone-induced changes in UCP3 mRNA expression are related to directs effects of T3 or reflect secondary effects of the hormone through stimulation of renin-angiotensin or -adrenergic systems. Hyperthyroidism was produced by injections of T3 with or without concomitant treatment with either captopril, propranolol or clenbuterol. T3 resulted in a significant increase in the relative abundance of UCP3 in heart and skeletal muscle (P < 0.05), and the effect was not altered by captopril or propanolol. There was no synergistic or additive effect of T3 and clenbuterol on UCP3 mRNA expression in skeletal muscle. We conclude that the effect of T3 on UCP3 expression in cardiac and skeletal muscle is not dependent on either angiotensin II or the -adrenergic system and probably reflects a direct action of the hormone on UCP3 gene expression...

Bioequivalence study of rabeprazole sodium on healthy human volunteers.

The newly developed proton pump inhibitor rabeprazole sodium is expected to have beneficial effects in the treatment of peptic ulcer. The pharmacokinetic parameters (C(max), AUC(o-t), t(max)) of this drug have been evaluated to compare the single dose (20 mg) bioavailability of rabeprazole sodium with the standard reference. High performance liquid chromatography (HPLC) coupled with UV detector set at 280 nm has been used to determine plasma concentration of 12 human volunteers as per Drugs Controller General of India (DCGI) guidelines. The method has been validated over a linear range of 20-480 ng/ml from plasma. The minimum quantifiable concentration was set at 10 ng/ml [co-efficient of variance (CV) < 10%]. By comparing AUC(o-t) the relative bioavailability of test preparation has been found to be 100.88% of that of reference preparation.

Effect of nitric oxide on H+ -efflux in presence of various nutrients in Candida albicans.

In the present study tentative link has been established between H+ -efflux and effect of NO in presence of various nutrients (glucose, 2-deoxy-D-glucose, xylose, proline, glutamic acid and lysine) in C. albicans using sodium nitroprusside (SNP) as a potent source of NO. It was observed that there was a decreasing trend in pH with time, in control, while SNP treated cells showed an initial decline in pH for 10-15 min, followed by an increase in pH up to 30 min. In presence of glucose there was an enhancement in H+ -efflux by 9-fold whereas proline, glutamic acid and lysine showed enhancement by 3, 6 and 1.5-fold respectively. Similar trends in increase in pH after 15 min in SNP treated cells of Candida was observed in presence of all nutrients used. It was demonstrated for the first time that H+ -ATPase of C. albicans was affected by NO.

Expression of an Anion Exchanger Pendrin in Human Kidney / 대한해부학회지

It has been reported that new apical anion exchanger perndrin, encoded by the pendred syndrome (PDS/pds, Slc26A4) gene, was expressed in the AE1-negative intercalated cells of rat and mouse kidneys. The purpose of this study was performed that expression of pendrin in the subtypes of intercalated cells in human kidney. The normal human renal tissues obtained from nephrotomized kidneys for renal cell carcinoma were fixed in periodate-lysine-paraformalde-hyde, and processed for immunohistochemistry. Subtypes of intercalated cells were identified by using antibodies for H(+)-ATPase and AE1, and connecting tubule cells and principal cells of collecting duct were identified using antibodies for calbindin D28K and AQP2, respectively. In human kidney, pendrin was expressed in the apical domain of AE1-negative intercalated cells including type B cells with diffuse and/or basolateal H(+)-ATPase, non A-non B (non -A/B) type intercalated cells with apical H(+)-ATPase and bipolar type of intercalated cells with apical and basolateral H(+)-ATPase. The AQP2-positive principal cells of cortical collecting duct were also had apical pendrin immunoreactivity. However, there was no pendrin immunoreactivity in AE1-positive type A intercalated cells, calbindin D28K-positive connecting tubule cells, and AQP2-positive medullary collecting duct. These results suggest that pendrin is an apical anion exchanger not only in the AE1-negative intercalated cells (type B, non-A/B and bipolar cells) but also in the principal cells of cortical collecting duct, and has an essential role in HCO3-secretion in human kidney.

Significance of Non-erosive Minimal Esophageal Lesions in Gastro-esophageal Reflux Disorder

BACKGROUND: Non-erosive reflux disorder, which represents more than 60% of gastro-esophageal reflux disorders, lacks objective parameters for diagnosis. The purpose of this study was to evaluate the correlation between non-erosive minimal lesions at the lower esophagus and gastro-esophageal reflux disorder. METHODS: Patients were asked to answer a symptom questionnaire. The endoscopic findings were either graded by LA classification or recorded as non-erosive minimal lesions. Patients with minimal lesions were treated with rabeprazole or a placebo and responses were evaluated at weeks 1 and 4. RESULTS: In 8 centers, 3454 patients were screened. In patients with heartburn or acid regurgitation as the most bothersome symptom, 23.7% had endoscopy negative reflux disorder, 40.1% showed minimal lesions, and 36.2% had mucosal break esophagitis. Thirty-four percent of patients with minimal lesions and 39.1% of patients with LA 'grade A' mild esophagitis reported typical reflux symptoms as their main symptom. In patients with minimal lesions, medication with rabeprazole reduced symptoms significantly at weeks 1 and 4, but not with the placebo. CONCLUSION: Patients with non-erosive minimal esophageal lesions had similar reflux symptoms comparable to those with mild erosive reflux esophagitis, and reflux symptoms were improved with a short-term proton pump inhibitor. Thus, non-erosive minimal esophageal lesion constitutes a great part of gastro-esophageal reflux disorder.

Expression of H(+)-ATPase in Inner Medullary Collecting Duct Cells of Aquaporin-3-Deficient Mice / 대한해부학회지

It has been reported that the decrease in urinary pH observed in AQP1 null mice with a urinary concentrating defect is due to upregulation of H(+)-ATPase in the IMCD. This is thought to be caused by the chronically low interstitial osmolality in these animals. To explore whether increase of H(+)-ATPase expression in the IMCD is associated with changes in the prolonged decrease of interstitial osmolality, we examined the expression of H(+)-ATPase and Na(+)-H(+) exchanger (NHE3) using light and electron microscopic immunocytochemistry in the kidneys of AQP3 null mice which are polyuric and manifest a urinary concentrating defect because of an inability to create a hypertonic medullary interstitium. In both AQP3 (-/-) and AQP1 (-/-) mouse kidneys, type A intercalated cells in cortical and medullary collecting ducts are slightly activated, and strong H(+)-ATPase immunostaining was present in the apical plasma membrane of IMCD cells, whereas no H(+)-ATPase labeling was observed in IMCD cells in wild type mice. No differences of the immunoreactivity for NHE3 in the proximal tubule and thick ascending limb of loop of Henle were observed between AQP3 or AQP1 (-/-) mice and AQP3 (+/+) mouse. These results suggest that the induction of H(+)-ATPase expression in IMCD cells of AQP3 null mice, as well as AQP1 null mice, may be related to their chronically low interstitial osmolality.

Changes of proton transportation across the inner mitochondrial membrane and H(+)-ATPase in endotoxic shock rats / 中华创伤杂志(英文版)

<p><b>OBJECTIVE</b>To investigate the changes of proton transportation across the inner mitochondrial membrane (IMM) and H(+)-ATPase of hepatocytes in endotoxic shock rats.</p><p><b>METHODS</b>Endotoxin from E. Coil of 5.0 mg/kg or saline of 1 ml/kg was injected into the femoral vein. The rats were sacrificed pre-injection and 1, 3, 5, 8 hours after injection, and plasma and liver tissue samples were collected respectively. The liver tissue samples were used for preparation of mitochondria and submitochondrial particles (SMPs). The proton-translocation of SMPs and H(+)-ATPase, phospholipase A(2) (PLA(2)) activities and malondialdehyde (MDA) content, membrane fluidities of different level of mitochondria membrane and plasma MDA content were assayed.</p><p><b>RESULTS</b>(1) Five hours after E. Coli. O111B4 injection, the maximum fluorescence quenching ACMA after adding ATP, nicotinamide adenin dinucleoacid hydrogen (NADH), and the succinate were significantly decreased (P<0.05). The time of maximum fluorescent quenching and the half time of fluorescent quenching were significantly prolonged (P<0.01), especially when NADH was used as a substrate. (2) The mitochondrial H(+)-ATPase activity was significantly increased at early stage of endotoxic shock (P<0.05), and significantly decreased at late stage of endotoxic shock (P<0.01). (3) The mitochondrial membrane bound PLA(2) activity, plasmal and mitochondrial MDA content were significantly increased and succinate dehydrogenase (SDH) activity of mitochondria decreased markedly in endotoxic shock rats (P<0.05). (4) The mitochondrial membrane fluidity of different lipid regions was decreased, especially in the head of phospholipid.</p><p><b>CONCLUSIONS</b>Proton transportation across IMM and mitochondrial H(+)-ATPase activity are significantly decreased in endotoxic shock.</p>