Expression of F0 ATP synthase C subunit in serum of patients with ischemic heart disease and its clinical implications / 中华危重病急救医学

Objective:To investigate the expression and clinical significance of F0 ATP synthase C subunit (Csub) in patients with ischemic heart disease (IHD).Methods:The 101 patients with chest pain admitted to the department of emergency of the People's Hospital of Yuhuan from May 2019 to December 2020 were enrolled, including 59 patients with acute myocardial infarction (AMI) and 42 patients with unstable angina pectoris (UAP). At the same time, 50 age-matched healthy subjects in the health examination center were selected as the healthy control (HC). All patients had completed blood sampling before the intervention of drugs or other intervention measures in the emergency room. The content of serum Csub was detected by enzyme linked immunosorbent assay (ELISA), and the relationship between Csub and clinical characteristics was analyzed. At the same time, the contents of hypersensitivity cardiac troponin T (hs-cTnT) and MB isoenzyme of creatine kinase (CK-MB) in blood were detected by electrochemical luminescence. The receiver operator characteristic curve (ROC curve) was drawn to evaluate the value of Csub, hs-cTnT, and CK-MB in the early diagnosis of IHD.Results:The baseline data such as age, gender, and history of the three groups were balanced. There were significant differences in low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), CK-MB, hs-cTnT and B-type natriuretic peptide (BNP), but there were no significant differences in other biochemical indexes. The Csub content in the AMI group and the UAP group were significantly higher than those in the HC group [8.96% (6.37%, 11.53%), 4.27% (3.23%, 6.49%) vs. 1.56% (1.07%, 2.33%), both P < 0.01]. Moreover, the Csub in the AMI group with more severe myocardial ischemia was higher than UAP group [8.96% (6.37%, 11.53%) vs. 4.27% (3.23%, 6.49%), P < 0.01]. A total of 59 patients with AMI were treated with percutaneous coronary intervention (PCI). According to the median of Csub, AMI patients were subdivided into above-median group (29 cases) and below-median group (30 cases). The results showed that there were no significant differences in the number of coronary artery lesion branches, the number of stent implantation and postoperative medication between the two groups. ROC curve analysis showed that the area under the curve (AUC) and 95% confidence interval (95% CI) of Csub, hs-cTnT and CK-MB in the diagnosis of IHD were 0.98 (0.95-1.00), 0.99 (0.99-1.00), 0.94 (0.89-0.99), respectively. The diagnostic efficacy of Csub was slightly lower than that of hs-cTnT but higher than that of CK-MB. When the cut-off value of Csub was 4.74%, the sensitivity and specificity for the diagnosis of IHD were 100% and 87.0%, respectively. Conclusions:Csub increased significantly in the serum of IHD patients, and further increased with the severity of ischemia. It can be used as a new diagnostic biomarker for the diagnosis and evaluation of the development of myocardial ischemia.

Development of KASP marker for cytoplasmic male sterility in Nicotiana tabacum and utilization in trait introgression

Cytoplasmic male sterility (CMS) is an important trait for large-scale hybrid seed production which avoids manual emasculation and undesired horizontal spread of pollen.Rearrangements in mitochondrial genome in terms of deletions and insertions are frequent causes leading to CMS. Mitochondrial ATP synthase is a multisubunit molecular machine which is involved in synthesis of ATP. In this study, three mutations in ATPase subunit 6 were identified and their cosegregation with male sterility was established using tobacco male sterile hybrids and Nicotiana suvaolensis. A breeder friendly Kompetitive allele specific polymerase chain reaction (KASP) SNP marker was developed for high throughput and quick genotyping. Introgression of this trait into selected germplasm lines (n = 9) was achieved based on foreground for CMS and background selection for recurrent parent using KASP marker and 50K custom tobacco SNP array, respectively. Analysis of genotyping data from SNP array revealed the presence of 88–99% of recurrent parent genome in BC3F1 plants. The selected BC3F1 plants exhibit CMS and are indistinguishable from the fertile recurrent parent (germplasm) in terms of plant morphology.

Prokaryotic expression of ATP5B and preparation and identification of its monoclonal antibody / 吉林大学学报(医学版)

Objective: To construct and identify the monoclonal antibody of ATP synthase beta subunit (ATP5B) with high purity, and to lay foundation for further study. Methods: The ATP5B gene was amplified by PCR and cloned into the pET28a vector and transformed into K. coli BL21 (DE3). The protein expression was induced by IPTG and then the fusion protein was purified by nickel affinity chromatography column. The protein purity was detected by SDS-polyacrylamide gel electrophoresis (SDS-PAGE). Three female Balb/C mice were immunized with purified fusion protein and the tail vein blood was taken to detect the titer of ATP5B antibody by indirect enzyme-linked immunosorbent assay (ELISA). The spleen cells from the immunized mice with the highest serum titer were mixed with the SP2/0 cells to establish the hybridoma cells and the fused cells were screened by indirect ELISA and monoclonally cultured. Karyotype analysis were performed in the positive cells. The hybridoma cells were intraperitoneally injected into 12 weeks old BALB/C mice to estabilish the ascites models. The titer of ascites was detected by indirect ELISA. The purity of tlie antibody was detected by SDS-PAGE. The antibody subtype was detected by ELISA. Results: After PCR amplification, a specific band of 1 455 bp was obtained, and the pET28a empty vector was ligated to obtain a recombinant pET28a/ATP5B vector. The target protein was expressed in the IPTG-induced bacteria solution; the SDS-PAGE results showed that the protein band was found at 51 000. The indirect ELISA results showed that the serum titer of the venous blood of immunized mice was up to 1: 64 000. In karyotype analysis, the total number of chromosomes in hybridoma cells was about the sum of myeloma cells and normal mouse spleen cells. The mouse ascites was prepared with the hybridoma cell line, and the highest titer of tlie antibody was 1 5 240 000. The subtype of the monoclonal antibody produced by the hybridoma cells was IgGl. Conclusion: The monoclonal antibody against ATP5B protein is successfully prepared by cloning, expressing and purifying the recombinant protein.

RNA editing analysis of ATP synthase genes in the cotton cytoplasmic male sterile line H276A

BACKGROUND: Pollen development is an energy-consuming process that particularly occurs during meiosis. Low levels of adenosine triphosphate (ATP) may cause cell death, resulting in CMS (cytoplasmic male sterility). DNA sequence differences in ATP synthase genes have been revealed between the N- and S-cytoplasms in the cotton CMS system. However, very few data are available at the RNA level. In this study, we compared five ATP synthase genes in the H276A, H276B and fertile F1 (H276A/H268) lines using RNA editing, RNA blotting and quantitative real time-PCR (qRT-PCR) to explore their contribution to CMS. A molecular marker for identifying male sterile cytoplasm (MSC) was also developed. RESULTS: RNA blotting revealed the absence of any novel orf for the ATP synthase gene sequence in the three lines. Forty-one RNA editing sites were identified in the coding sequences. RNA editing showed that proteins had 32.43% higher hydrophobicity and that 39.02% of RNA editing sites had proline converted to leucine. Two new stop codons were detected in atp6 and atp9 by RNA editing. Real-time qRT-PCR data showed that the atp1, atp6, atp8, and atp9 genes had substantially lower expression levels in H276A compared with those in H276B. By contrast, the expression levels of all five genes were increased in F1 (H276A/H268). Moreover, a molecular marker based on a 6-bp deletion upstream of atp8 in H276A was developed to identify male sterile cytoplasm (MSC) in cotton. CONCLUSIONS: Our data substantially contributes to the understanding of the function of ATP synthase genes in cotton CMS. Therefore, we suggest that ATP synthase genes might be an indirect cause of cotton CMS. Further research is needed to investigate the relationship among ATP synthase genes in cotton CMS.

Prokaryotic expression of ATP5B and preparation and identification of its monoclonal antibody / 吉林大学学报(医学版)

Objective:To construct and identify the monoclonal antibody of ATP synthase beta subunit (ATP5B) with high purity, and to lay foundation for further study.Methods:The ATP5Bgene was amplified by PCR and cloned into the pET28avector and transformed into E.coli BL21 (DE3) .The protein expression was induced by IPTG and then the fusion protein was purified by nickel affinity chromatography column.The protein purity was detected by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) .Three female Balb/C mice were immunized with purified fusion protein and the tail vein blood was taken to detect the titer of ATP5Bantibody by indirect enzyme-linked immunosorbent assay (ELISA) .The spleen cells from the immunized mice with the highest serum titer were mixed with the SP2/0cells to establish the hybridoma cells and the fused cells were screened by indirect ELISA and monoclonally cultured.Karyotype analysis were performed in the positive cells.The hybridoma cells were intraperitoneally injected into 12weeks old BALB/C mice to estabilish the ascites models.The titer of ascites was detected by indirect ELISA.The purity of the antibody was detected by SDS-PAGE.The antibody subtype was detected by ELISA.Results:After PCR amplification, a specific band of 1 455bp was obtained, and the pET28aempty vector was ligated to obtain a recombinant pET28a/ATP5Bvector.The target protein was expressed in the IPTG-induced bacteria solution;the SDS-PAGE results showed that the protein band was found at51 000.The indirect ELISA results showed that the serum titer of the venous blood of immunized mice was up to1:64 000.In karyotype analysis, the total number of chromosomes in hybridoma cells was about the sum of myeloma cells and normal mouse spleen cells.The mouse ascites was prepared with the hybridoma cell line, and the highest titer of the antibody was 1:240 000.The subtype of the monoclonal antibody produced by the hybridoma cells was IgG1.Conclusion:The monoclonal antibody against ATP5Bprotein is successfully prepared by cloning, expressing and purifying the recombinant protein.

Preliminary identification of the outer membrane protein H receptors of avian Pasteurella multocida / 中国人兽共患病学报

To identify the receptors for the outer membrane protein H (OmpH) of avian P.multocida,the membrane proteins of chicken embryo fibroblast (CEF) cells were separated by SDS-PAGE and analyzed by Ligand blot.The OmpH-binding protein was identified by MALDI-TOF mass spectrometry,and its distribution in the membrane proteins of different host esophageal mucosal cells was detected by Ligand blot,ELISA and immunofluorescence microscopy,respectively.Ligand blot analysis showed that a 49-kDa membrane protein of CEF cells bound to recombinant OmpH,and MALDI-TOF spectral results demonstrated that the OmpH-binding protein was ATP synthase β subunit.In addition,the OmpH receptor was present in the chicken and rabbit mucosal cell membranes,but was not detected in the bovine and swine mucosal cell membranes.The above results indicate that the OmpH receptor may be CEF cell-derived ATP synthase beta subunit.

ATP Synthase β-subunit Abnormality in Pancreas Islets of Rats with Polycystic Ovary Syndrome and Type 2 Diabetes Mellitus / 华中科技大学学报（医学）（英德文版）

This study investigated the abnormal expression of ATP synthase β-subunit (ATPsyn-β) in pancreas islets of rat model of polycystic ovary syndrome (PCOS) with type 2 diabetes mellitus (T2DM),and the secretion function changes after up-regulation of ATP5b.Sixty female SD rats were divided into three groups randomly and equally.The rat model of PCOS with T2DM was established by free access to the high-carbohydrate/high-fat diet,subcutaneous injections of DHEA,and a single injection of streptozotocin.The pancreas was removed for the detection of the ATPsyn-β expression by immunohistochemical staining,Western blotting and reverse transcription-PCR (RT-PCR).The pancreas islets of the rats were cultured,isolated with collagenase Ⅴ and purified by gradient centrifugation,and the insulin secretion after treatment with different glucose concentrations was tested.Lentivirus ATP5b was successfully constructed with the vector of GV208 and transfected into the pancreas islets for the over-expression of ATPsyn-β.The insulin secretion and intracellular ATP content were determined after transfection of the PCOS-T2DM pancreas islets with Lenti-ATP5b.The results showed that the expression of ATPsyn-β protein and mRNA was significantly decreased in the pancreas of PCOS-T2DM rats.The ATP content in the pancreas islets was greatly increased and the insulin secretion was improved after the up-regulation of ATPsyn-β in the pancreas islets transfected with lenti-ATP5b.These results indicated that for PCOS,the ATPsyn-β might be one of the key factors for the attack of T2DM.

Research progress on the mitochondrial respiratory chain complex Ⅴ in sepsis / 国际儿科学杂志

Sepsis is a complex pathophysiological disorder arising from a systemic inflammatory response to infection.It is considered as a major cause of death among critically ill patients in intensive care units.Mitochondrial dysfunction and impaired oxidative phosphorylation have been implicated in the pathogenesis of sepsis.The mitochondrial respiratory chain complex Ⅴ is the key enzyme of oxidative phosphorylation.It is response for ATP synthesis and there is a reduction of complex Ⅴ activity in the early sepsis.However,in the late sepsis,under ischemic conditions, it accounts for significant ATP hydrolysis and the activity is up-regulation.Here,this review focuses on the changes of complex Ⅴ activity,mechanism of regulation and therapy in sepsis.

Expression of hepatic energy proteins following reduced-size liver transplantation in rats / 中国组织工程研究

BACKGROUND:At present, the proteome is a mature technology that has been applied in basic research fields related to liver transplantation. But, it has been not reported in research related to reduced-size liver transplantation. OBJECTIVE:To explore the expression of differential proteins related to hepatic energy metabolism fol owing reduce-size liver transplantation in rats by using by proteomic technology. METHODS:The improved model of reduced-size liver transplantation was used in this experiment. The donor was health female Lewis rats and the recipient was male Wistar rats for liver transplantation. The difference between the donor and the recipient was about 20 g. The weight of donor liver/the weight of recipient donor was approximately equal to 50%. The donor liver tissue was harvested and trimmed to the required size. The portal vein and infrahepatic vena cava were cannulated, and the biliary tract was implanted into the donor bile duct for transplantation. Then the donor was transplanted into the recipient after the removal of original liver tissue. Hepatic specimens were harvested by 1, 3 and 7 days after reduced-size liver transplantation. Then, the harvested specimens were compared with the normal donor and recipient liver tissue that were previously harvested and frozen, to generate two-dimensional gel electrophoresis profile using proteome technology. Then tandem mass spectrometry and databases analysis were performed after two-dimensional electrophoresis for identifying differential protein stains. RESULTS AND CONCLUSION:In this experiment, 72 differential protein stains with over lo-fold changes were selected. After identification, 32 proteins showed clear functions, and among them three differential proteins (ATP synthase beta subunit, electron-transferring flavoprotein beta peptide and proton-transferring ATP synthase) were involved in the process of cel energy metabolism. The proteins were distributed on 1 and 7 days after reduce-size liver transplantation, accounting for 6%.

Effect of Combined Aerobic and Anaerobic Training on Hemodynamic Response, Serum Oxidative Stress Markers and Platelet ATP Synthase Activities in Patients with Coronary Heart Disease: A Pilot Study.

Background: The overall aim of this work was to study the impact of combined aerobic and anaerobic training in relation to hemodynamic response (heart rate, systolic blood pressure, and double product), serum oxidative stress markers (lipoperoxides, nitrites-nitrates) and platelet ATP synthase activities in patients with coronary heart disease. Materials and Methods: Ten subjects, 9 male and 1 female, (mean age 57.7±7.2 years) with coronary heart disease participated in this study. Patients performed combined aerobic and anaerobic exercise for 24 sessions (three times a week). Results: The results suggest myocardium adaptations, manifested in the reduction of heart rate with increased workloads and increased double product [(heart rate) x (systolic blood pressure)] according to the intensity, frequency and duration of training. The ATP synthesis rate was significantly increased at session 3 (post-exercise) compared to session 1 (pre exercise). Furthermore, rate of ATP hydrolysis was significantly decreased at session 24 (post-exercise 3) compared to session 1 (post-exercise 1). Serum lipid peroxidation products and nitric oxide catabolites were significantly diminished at session 24 (pre-exercise). Conclusion: In some patients hemodynamic responses showed improvements in response to exercise. The exercise sessions induced lower levels of lipid peroxidation products, nitric oxide catabolites and ATPase activity. Conversely, ATP synthase activity showed higher values at the end of the experiment. These results confirm the beneficial effect of combined aerobic and anaerobic exercise.

The inhibitory effects of rotenone, 2, 4-dinitrophenol, oligomycin on the electron proton flow through “The membrane redox potential three state line” / Монголын Анагаах Ухаан

Rotenone is a specific inhibitor of the NADH dehydrogenase complex. In mitochondria, rotenone inhibitsthe oxidation of NADH to NAD, thereby blocking the oxidation of NAD and the substrates such asglutamate, alpha-ketoglutarate, and pyruvate. Rotenone also inhibits the mitochondrial respiratory chainbetween diphosphopyridine nucleotide and flavine.2, 4-Dinitrophenol – (DNP) is lipophilic weak acids that pick up a proton, transport across the mitochondrialinner membrane into the matrix, deprotonate, then exit as anions before repeating the catalytic cycle,and dissipating the proton gradient. In this situation, electrons continue to pass through the electrontransport system, reduce oxygen to water and metabolic rate, heat are increased, but ATP is lesssynthesized in this process.The macrolide antibiotic - oligomycin binds to the surface of the c8-10 ring of the Fo domain of ATPsynthase, making contact with two neighboring molecules and blocking proton flow, which explains theinhibitory effect on ATP synthesis. Intraperitoneal injection of oligomycin into the rat (0.5 mg per kg)reduces the oxygen consumption by about 50%; decreases ATP production by the aerobic pathway andincreases formation of lactate in blood serum. These changes may cause a decelerated metabolism andan increased formation of free radicals or ROS in membranes.

White Spot Syndrome Virus infection in Penaeus monodon is facilitated by housekeeping molecules.

White Spot Syndrome Virus (WSSV) is a major pathogen in shrimp aquaculture, and its rampant spread has resulted in great economic loss. Identification of host cellular proteins interacting with WSSV will help in unravelling the repertoire of host proteins involved in WSSV infection. In this study, we have employed one-dimensional and twodimension virus overlay protein binding assay (VOPBA) followed by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) to identify the host proteins of Penaeus monodon that could interact with WSSV. The VOPBA results suggest that WSSV interacted with housekeeping proteins such as heat shock protein 70, ATP synthase subunit β, phosphopyruvate hydratase, allergen Pen m 2, glyceraldehyde-3-phosphate dehydrogenase, sarcoplasmic calcium-binding protein, actin and 14-3-3-like protein. Our findings suggest that WSSV exploits an array of housekeeping proteins for its transmission and propagation in P. monodon.

Research advances in cell surface ATP synthase / 上海第二医科大学学报

Mitochondria ATP synthase is a key enzyme in cellular energy interconversion. It is generally believed that ATP synthase is strictly confined to mitochondria. However, it has been demonstrated that ATP synthase also occurs on the extracellular surface of vascular endothelial cells, tumor cells and adipocytes, instead of normal cells. Based on this characteristic, many functional researches have been conducted on the angiogenesis, tumor inhibition and lipid metabolism, and the mechanism on which this enzyme works has been preliminarily elucidated. The research advances in cell-surface ATP synthase are reviewed in this paper.

Secretion of adenylate kinase 1 is required for extracellular ATP synthesis in C2C12 myotubes

Extracellular ATP (exATP) has been known to be a critical ligand regulating skeletal muscle differentiation and contractibility. ExATP synthesis was greatly increased with the high level of adenylate kinase 1 (AK1) and ATP synthase beta during C2C12 myogenesis. The exATP synthesis was abolished by the knock-down of AK1 but not by that of ATP synthase beta in C2C12 myotubes, suggesting that AK1 is required for exATP synthesis in myotubes. However, membrane-bound AK1beta was not involved in exATP synthesis because its expression level was decreased during myogenesis in spite of its localization in the lipid rafts that contain various kinds of receptors and mediate cell signal transduction, cell migration, and differentiation. Interestingly, cytoplasmic AK1 was secreted from C2C12 myotubes but not from C2C12 myoblasts. Taken together all these data, we can conclude that AK1 secretion is required for the exATP generation in myotubes.

Extracellular ATP is generated by ATP synthase complex in adipocyte lipid rafts

Mitochondrial biogenesis is known to accompany adipogenesis to complement ATP and acetyl-CoA required for lipogenesis. Here, we demonstrated that mitochondrial proteins such as ATP synthase alpha and beta, and cytochrome c were highly expressed during the 3T3-L1 differentiation into adipocytes. Fully-differentiated adipocytes showed a significant increase of mitochondria under electron microscopy. Analysis by immunofluorescence, cellular fractionation, and surface biotinylation demonstrated the elevated levels of ATP synthase complex found not only in the mitochondria but also on the cell surface (particularly lipid rafts) of adipocytes. High rate of ATP (more than 30 micrometer) synthesis from the added ADP and Pi in the adipocyte media suggests the involvement of the surface ATP synthase complex for the exracellular ATP synthesis. In addition, this ATP synthesis was significantly inhibited in the presence of oligomycin, an ATP synthase inhibitor, and carbonyl cyanide m-chlorophenylhydrazone (CCCP), an ATP synthase uncoupler. Decrease of extracellular ATP synthesis in acidic but not in basic media further indicates that the surface ATP synthase may also be regulated by proton gradient through the plasma membrane.

Expression of ATP synthase 6 in breast cancer cell line, T47D cells, induced by heregulin β1 / 吉林大学学报(医学版)

Objective : To explore expression of related genes in breast cell line T47D induced by heregulin β1. Methods: Suppression subtractive hybridization was performed using cDNA sub traction kit to detect expression of heregulin-responsive genes in T47D cells. Results: ATP syn thase 6 was up-regulated by heregulin β1 in T47D cells at 1 and 6 hours. Conclusion: Heregulin β1 participates in the regulation of expression of both ATP synthase 6 and oxidative phosphoryla tion of T47D cells.

Vesicular transport as a new paradigm in short-term regulation of transepithelial transport

The vectorial transepithelial transport of water and electrolytes in the renal epithelium is achieved by the polarized distribution of various transport proteins in the apical and basolateral membrane. The short-term regulation of transepithelial transport has been traditionally thought to be mediated by kinetic alterations of transporter without changing the number of transporters. However, a growing body of recent evidence supports the possibility that the stimulus-dependent recycling of transporter-carrying vesicles can alter the abundance of transporters in the plasma membrane in parallel changes in transepithelial transport functions. The abundance of transporters in the plasma membrane is determined by net balance between stimulus-dependent exocytic insertion of transporters into and endocytic retrieval of them from the plasma membrane. The vesicular recycling occurs along the tracts of the actin microfilaments and microtubules with associated motors. This review is to highlight the importance of vesicular transport in the short-term regulatory process of transepithelial transport in the renal epithelium. In the short-term regulation of many other renal transporters, vesicular transport is likely to be also involved. Thus, vesicular transport is now emerged as a wide-spread general regulatory mechanism involved in short-term regulation of renal functions.

Mitochondrial ATP synthase genes may be implicated in cytoplasmic male sterility in Sorghum bicolor.

Incompatible nuclear-cytoplasmic interactions are responsible for the phenomenon of cytoplasmic male sterility in plants. We have analysed male sterile (2077A, 296A), maintainer fertile (2077B, 296B) and fertility restored (2077R, 296R) lines of sorghum for the restriction fragment locations of various mitochondrial genes and their transcripts. We report here a polymorphism in genes related to the ATP synthase complex between two different cytoplasms from the A and Β set of lines of 2077 and 296. There is also a difference in the transcript size of the atpA gene between the A and Β cytoplasms. We propose that incompatibility in nuclear cytoplasmic interactions may be explained in terms of incompatible subunits being synthesized by the mitochondria and nucleus for a multisubunit complex of the mitochondrial membrane such as ATPase.

Effects of hyperbaric oxygen on cerebral mitochondrial energy metabolism after brain ischemia/reperfusion in rats / 重庆医科大学学报

Objective:To study the effects of hyperbaric oxygen(HBO) on the levels of cerebral mitochondrial adenosine triphosphate (ATP) and proton-translocating ATP synthases(F 0F 1-ATPase) after brain ischemia/reperfusion (IR) in rats.Methods:Thirty Wistar rats were randomly divided into 3 groups:normal contorl group (n=6) ,IR group (n=12) and HBO group ( n=12 ).IR was performed by ligation of the middle cerebral artery for 2 hours (hs) and then reperfusion in both the IR and HBO groups.The IR group was subdivided into group I 2h R 2h and group I 2h R 24h .The HBO group was subdivided into group HBO-I 2h R 2h and group HBO-I 2h R 24h .The IR group was treated by 2.5 ATAO 2.The HBO group was treated by 2.5 ATA O 2.The pure mitochondria was separated by Clak's Ficoll density gradient method.The activity of F 0F 1-ATPase was determined by biochemical technigue and the level of cerebral ATP by high-performance liquid chromatography (HPLC).Results:The level of ATP and the activity of F 0F 1-ATPase increased after HBO treatment.In the IR group,the level of ATP and the activity of F 0F 1-ATPase were significantly lower than those of the HBO group( P