Role of ROS/Kv/HIF Axis in the Development of Hypoxia-Induced Pulmonary Hypertension

Hypoxic pulmonary hypertension (HPH) is a common complication in patients with chronic obstructive pulmonary disease (COPD), sleep-disordered breathing, or dwellers in high altitude. The exact mechanisms underlying the development of HPH still remain unclear. Reactive oxygen species (ROS),hypoxia inducible factors (HIF), and potassium channels (KV) are believed as the main factors during the development of HPH. We propose that the “ROS/Kv/HIF axis” may play an important initiating role in the development of HPH. Being formed under a hypoxic condition, ROS affects the expression and function of HIFs or KV, and consequently triggers multiple downstream signaling pathways and genes expression that participate in promoting pulmonary vasoconstriction and arterial remodeling. Thus, further study determining the initiating role of “ROS/Kv/HIF axis” in the development of HPH could provide theoretic evidences to better understand the underlying mechanisms of HPH, and help identify new potential targets in the treatment of HPH.

Hypoxia and cytokines regulate carbonic anhydrase 9 expression in hepatocellular carcinoma cells in vitro

AIM: To study the expression of carbonic anhydrase(CA) 9 in human hepatocellular carcinoma(HCC) cells.METHODS: We studied CA9 protein, CA9 m RNA and hypoxia-inducible factor-1 alpha(HIF-1α) protein levels in Hep3 B cells exposed in different parallel approaches. In one of these approaches, HCC cells were exposed to extreme in vitro hypoxia(24 h 0.1% O2) without or with interleukin(IL)-1, IL-6, tumor necrosis factoralpha(TNF-α) and transforming growth factor-beta(TGF-β) stimulation for the same hypoxic exposure time or exposed to normoxic oxygenation conditions without or with cytokine stimulation.RESULTS: The tumour cell line analysed showed a strong hypoxic CA9 m RNA expression pattern in response to prolonged severe hypoxia with cell-line specific patterns and a marked induction of CA9 protein in response to severe hypoxia. These results were paralleled by the results for HIF-1α protein under identical oxygenation conditions with a similar expression tendency to that displayed during the CA9 protein expression experimental series. Continuous stimulation with the cytokines, IL-1, IL-6, TNF-α and TGF-β, under normoxic conditions significantly increased the carbonic anhydrase 9 expression level at both the protein and m RNA level, almost doubling the CA9 m RNA and CA9 and HIF-1α protein expression levels found under hypoxia. The findings from these experiments indicated that hypoxia is a positive regulator of CA9 expression in HCC, and the four signal transduction pathways, IL-1, IL-6, TNF-α and TGF-β, positively influence CA9 expression under both normoxic and hypoxic conditions.CONCLUSION: These findings may potentially be considered in the design of anti- cancer therapeutic approaches involving hypoxia-induced or cytokine stimulatory effects on expression. In addition, they provideevidence of the stimulatory role of the examined cytokine families resulting in an increase in CA9 expression under different oxygenation conditions in human cancer, especially HCC, and on the role of the CA9 gene as a positive disease regulator in human cancer.

Hypoxia regulates reactive oxygen species levels in SHG-44 glioma cells

In the present study, cultured human SHG-44 glioma cells were subjected to a hypoxic environment simulated using the CoOl2 method. Flow cytometry showed increased reactive oxygen species production in these cells. Real-time reverse transcription-PCR showed significantly increased hypoxia-inducible factor-la mRNA expression in cells exposed to the hypoxic condition. The antioxidant N-acetylcysteine significantly inhibited reactive oxygen species production and reduced hypoxia-inducible factor-la mRNA expression in normoxic and hypoxic groups, especially in the latter group. These findings indicate that hypoxia induces reactive oxygen species production and hypoxia-inducible factor-la mRNA expression in human SHG-44 glioma cells, and that the antioxidant N-acetylcysteine can inhibit these changes.

Rates of oxygen consumption and tolerance of hypoxia and desiccation in Chinese black sleeper(Bostrichthys sinensis)and mudskipper(Boleophthalmus pectinirostris)embryos

The rates of oxygen consumption, tolerance of hypoxia and desiccation of the Chinese black sleeper （ Bostrichthys sinensis） and mudskipper （Boleophthalmus pectinirostris） embryos were investigated. The pattern of oxygen consumption of the Chinese black sleeper embryos was similar to that of the mudskipper ones. The lowest rates of oxygen consumption [ （ 1.65 ±0. 66） nmol/ （ind. ·h） ] of the Chinese black sleeper embryos 16 h after fertilization and the lowest rates of oxygen consumption [ （0.79± 0.08 ）nmolf（ ind. · h） ] of the mudskipper embryos 6 h after fertilization were recorded, respectively. Then the rates of oxygen consumption of these two species embryos increased gradually until hatching [ （8.26 ± 1.70 ） nmolf（ ind.· h） in the Chinese black sleeper, （2.69 ± 0.23 ）nmolf（ ind. · h） in mudskipper]. After exposure to hypoxia water （0.16 mg/dm^3）, bradycardia of the embryos occurred in both the Chinese black sleeper and the mudskipper. However, the Chinese black sleeper embryos survived approximately 45 min longer than the mudskipper ones. After exposure to desiccation at a relative humidity of 58%, bradycardia of the embryos was observed in both the Chinese black sleeper and the mudskipper, and the Chinese black sleeper embryos lived approximately 9 min longer than the mudskipper ones.

The ROCK pathway inhibitor Y-27632 mitigates hypoxia and oxidative stress-induced injury to retinal Müller cells

Rho kinase （ROCK） was the first downstream Rho effector found to mediate RhoA-induced actin cytoskeletal changes through effects on myosin light chain phosphorylation. There is abundant evidence that the ROCK pathway participates in the pathogenesis of retinal endothelial injury and proliferative epiretinal membrane traction. In this study, we investigated the effect of the ROCK pathway inhibitor Y-27632 on retinal Müller cells subjected to hypoxia or oxidative stress. Müller cells were subjected to hypoxia or oxidative stress by exposure to CoCl2 or H2O2. After a 24-hour treatment with Y-27632, the 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyl tetrazolium bromide assay was used to assess the survival of Müller cells. Hoechst 33258 was used to detect apoptosis, while 2′,7′-dichlorodihydrofluorescein diacetate was used to measure reactive oxygen species generation. A transwell chamber system was used to examine the migration ability of Müller cells. Western blot assay was used to detect the expression levels of α-smooth muscle actin, glutamine synthetase and vimentin. After treatment with Y-27632, Müller cells subjected to hypoxia or oxidative stress exhibited a morphology similar to control cells. Y-27632 reduced apoptosis, α-smooth muscle actin expression and reactive oxygen species generation under oxidative stress, and it reduced cell migration under hypoxia. Y-27632 also upregulated glutamine synthetase expression under hypoxia but did not impact vimentin expression. These findings suggest that Y-27632 protects Müller cells against cellular injury caused by oxidative stress and hypoxia by inhibiting the ROCK pathway.

Macrobenthic assemblages of the Changjiang River estuary (Yangtze River, China) and adjacent continental shelf relative to mild summer hypoxia

To assess the effects of hypoxia, macrobenthic communities along an estuarine gradient of the Changjiang estuary and adjacent continental shelf were analyzed. This revealed spatial variations in the communities and relationships with environmental variables during periods of reduced dissolved oxygen （DO） concentration in summer. Statistical analyses revealed significant differences in macrobenthic community composition among the three zones： estuarine zone （EZ）, mildly hypoxic zone （MHZ） in the continental shelf, and norrnoxic zone （NZ） in the continental shelf（Global R=0.206, P=0.002）. Pairwise tests showed that the macrobenthic community composition of the EZ was significantly different from the MHZ （pairwise test R=0.305, P=0.001） and the NZ （pairwise test R=0.259, P=-0.001）. There was no significant difference in macrobenthic communities between the MHZ and the NZ （pairwise test R=0.062, P=-0.114）. The taxa included small and typically opportunistic polychaetes, which made the greatest contribution to the dissimilarity between the zones. The effects of mild hypoxia on the macrobenthic communities are a result not only of reduced DO concentration but also of differences in environmental variables such as temperature, salinity, and nutrient concentrations caused by stratification.

DI-3-n-butylphthalide exerts neuroprotective effects by modulating hypoxia-inducible factor 1-alpha ubiquitination to attenuate oxidative stress-induced apoptosis

DI-3-n-butylphthalide is used to treat mild and moderate acute ischemic stroke.However,the precise underlying mechanism requires further investigation.In this study,we investigated the molecular mechanism of DI-3-n-butylphthalide action by various means.We used hydrogen peroxide to induce injury to PC12cells and RAW264.7 cells to mimic neuronal oxidative stress injury in stroke in vitro and examined the effects of DI-3-n-butylphthalide.We found that DI-3-nbutylphthalide pretreatment markedly inhibited the reduction in viability and reactive oxygen species production in PC12 cells caused by hydrogen peroxide and inhibited cell apoptosis.Furthermore,DI-3-n-butylphthalide pretreatment inhibited the expression of the pro-apoptotic genes Bax and Bnip3.DI-3-nbutylphthalide also promoted ubiquitination and degradation of hypoxia inducible factor 1α,the key transcription factor that regulates Bax and Bnip3 genes.These findings suggest that DI-3-n-butylphthalide exhibits a neuroprotective effect on stroke by promoting hypoxia inducible factor-1α ubiquitination and degradation and inhibiting cell apoptosis.

Effects of dissolved oxygen and nutrients from the Kuroshio on hypoxia off the Changjiang River estuary

The intrusion of the Kuroshio into the East China Sea(ECS)aff ects the development of hypoxia off the Changjiang(Yangtze)River estuary;however,quantitative analysis of its impacts is lacking.In this study,the Regional Ocean Modeling Systems(ROMS)model coupled with the Carbon,Silicate and Nitrogen Ecosystem(CoSiNE)model was used to investigate the relative importance of dissolved oxygen(DO)and diff erent nutrients(silicate,nitrate,and phosphate)in the Kuroshio on hypoxia in the ECS.Results show that changes in DO concentrations in the Kuroshio modify the distribution and intensity of hypoxia through direct onshore transport by hydrodynamic processes.An increase in Kuroshio DO concentration by 25%or 50%would result in a decrease of the maximum hypoxia extent(MHE)in the ECS by 76%or 86%,respectively,while a 25%decrease in Kuroshio DO would increase the MHE by up to 219%.The contribution of DO in the Taiwan Strait is almost negligible.In contrast to Kuroshio DO,nutrients aff ect hypoxia in the ECS through onshore transport by hydrodynamic and biochemical processes.Changes in phosphate and nitrate concentrations by 25%in the Kuroshio would change the MHE by up to 30%and 18%,respectively,accompanied by apparent changes in surface chlorophyll-a concentrations.The eff ect of silicate on hypoxia is negligible because a 25%change in silicate concentrations in the Kuroshio would result in less than 1%change in the MHE.Our results reveal a hierarchical rank of importance for environmental variables in the Kuroshio(i.e.,DO>phosphate>nitrate>silicate)in modifying the development of hypoxia in the ECS.

Can hypobaric hypoxia affect human thermal comfort?An experimental study in Tibet,China

Hypobaric hypoxia is the main environmental feature of the Tibetan plateau which would influence the efficiency of human metabolic heat production and the ability of thermal regulation.In order to understand the influence of the hypoxic environment on the plateau on the thermal comfort of short-term sojourners in Tibet,China,oxygen generators were used to create oxygen-enriched environments,and physiological and psychological reactions of subjects were compared under different oxygen partial pressures(p_(O_(2)))and air temperatures(t_(a)).The results showed that subjects’thermal sensation,thermal comfort and mean skin temperature decreased with a decrease in the oxygen partial pressure.When t_(a)=17℃,the influence of oxygen partial pressure was more pronounced,compared to p_(O_(2))=16.4 kPa,the thermal sensation of subjects under p_(O_(2))=13.7 kPa decreased by 33%.The rate of subjects feeling comfortable decreased by 25%,and the mean skin temperature decreased by 0.7℃.The hypoxic environment of the plateau exacerbates human discomfort.Therefore,it is necessary to fully understand the actual thermal requirements of sojourners in Tibet,China.The results of this study would have implications for a better understanding of thermal comfort characteristics in the hypoxia environment in plateau.

Regulation of heme oxygenase expression by alcohol,hypoxia and oxidative stress

AIM:To study the effect of both acute and chronic alcohol exposure on heme oxygenases(HOs) in the brain,liver and duodenum.METHODS:Wild-type C57BL/6 mice,heterozygous Sod2 knockout mice,which exhibit attenuated manganese superoxide dismutase activity,and liver-specific ARNT knockout mice were used to investigate the role of alcohol-induced oxidative stress and hypoxia.For acute alcohol exposure,ethanol was administered in the drinking water for 1 wk.Mice were pair-fed with regular or ethanol-containing Lieber De Carli liquid diets for 4 wk for chronic alcohol studies.HO expression was analyzed by real-time quantitative polymerase chain reaction and Western blotting.RESULTS:Chronic alcohol exposure downregulated HO-1 expression in the brain but upregulated it in the duodenum of wild-type mice.It did not alter liver HO-1 expression,nor HO-2 expression in the brain,liver or duodenum.In contrast,acute alcohol exposure decreased both liver HO-1 and HO-2 expression,and HO-2 expression in the duodenum of wild-type mice.The decrease in liver HO-1 expression was abolished in ARNT+/-mice.Sod2+/-mice with acute alcohol exposure did not exhibit any changes in liver HO-1 and HO-2 expression or in brain HO-2 expression.However,alcohol inhibited brain HO-1 and duodenal HO-2 but increased duodenal HO-1 expression in Sod2+/-mice.Collectively,these findings indicate that acute and chronic alcohol exposure regulates HO expression in a tissue-specific manner.Chronic alcohol exposure alters brain and duodenal,but not liver HO expression.However,acute alcohol exposure inhibits liver HO-1 and HO-2,and also duodenal HO-2 expression.CONCLUSION:The inhibition of liver HO expression by acute alcohol-induced hypoxia may play a role in the early phases of alcoholic liver disease progression.

Sirtuin1 attenuates acute liver failure by reducing reactive oxygen species via hypoxia inducible factor 1α

BACKGROUND The occurrence and development of acute liver failure(ALF)is closely related to a series of inflammatory reactions,such as the production of reactive oxygen species(ROS).Hypoxia inducible factor 1α(HIF-1α)is a key factor that regulates oxygen homeostasis and redox,and the stability of HIF-1αis related to the ROS level regulated by Sirtuin(Sirt)family.The activation of Sirt1 will lead to a powerful antioxidant defense system and therapeutic effects in liver disease.However,little is known about the relationship between HIF-1αand Sirt1 in the process of ALF and the molecular mechanism.AIM To investigate whether HIF-1αmay be a target of Sirt1 deacetylation and what the effects on ALF are.METHODS Mice were administrated lipopolysaccharide(LPS)/D-gal and exposed to hypoxic conditions as animal model,and resveratrol was used as an activator of Sirt1.The cellular model was established with L02 cells stimulated by LPS.N-acetyl-Lcysteine was used to remove ROS,and the expression of Sirt1 was inhibited by nicotinamide.Western blotting was used to detect Sirt1 and HIF-1αactivity and related protein expression.The possible signaling pathways involved were analyzed by immunofluorescent staining,co-immunoprecipitation,dihydroethidium staining,and Western blotting.RESULTS Compared with mice stimulated with LPS alone,the expression of Sirt1 decreased,the level of HIF-1αacetylation increased in hypoxic mice,and the levels of carbonic anhydrase 9 and Bcl-2-adenovirus E1B interacting protein 3 increased significantly,which was regulated by HIF-1α,indicating an increase of HIF-1αactivity.Under hypoxia,the down-regulation of Sirt1 activated and acetylated HIF-1αin L02 cells.The inhibition of Sirt1 significantly aggravated this effect and the massive production of ROS.The regulation of ROS was partly through peroxisome proliferatoractivated receptor alpha or AMP-activated protein kinase.Resveratrol,a Sirt1 activator,effectively relieved ALF aggravated by hypoxia,the production of ROS,and cell apoptosis.It also induced the deacetylation of HIF-1αand inhibited the activity of HIF-1α.CONCLUSION Sirt1 may have a protective effect on ALF by inducing HIF-1α deacetylation to reduce ROS.

Propofol blocks apoptosis and Bcl-2 and Bax expression induced by hypoxia-reoxygenation in primary cultures of rat hippocampal astrocytes

BACKGROUND： Cerebral hippocampal astrocytes are more sensitive.to ischemic injury than neurons. Hypoxic-ischemic brain injury induces profound astrocyte apoptosis, and propofol may protect against astrocyte apoptosis. OBJECTIVE： To verify the protective effects of propofol against astrocyte apoptosis and to investigate anti-apoptotic Bcl-2 and pro-apoptotic Bax expression in primary cultures of rat hippocampal astrocytes exposed to hypoxia-reoxygenation for different periods of time following propofol treatment. DESIGN, TIME, AND SETTING： In vitro neural immunocytochemistry was performed at the Central Laboratory of Yunyang Medical College between September 2007 and March 2008.MATERIALS： A total of 30 Wistar rats, aged 1-3 days, wJth equal numbers of males and females, were included for isolation and culture of .hippocampal astrocytes. METHODS： Hippocampal astrocytes were purified and cultured for 3 weeks and treated with four culture conditions： 50 μL Hank＇s solution （normal control）; 0.2 mL/L Intralipid; 50 μL Hank＇s solution for 10 minutes followed by hypoxic incubation for 4 hours and normoxic incubation for 12, 24, 36, 48, 60 or 72 hours; propofol （250 μmol/L final） for 10 minutes followed by hypoxic incubation for 4 hours and normoxic incubation for 12, 24, 36, 48, 60 and 72 hours. MAIN OUTCOME MEASURES： （1） Morphologic changes in hippocampal astrocytes. （2） Levels of astrocyte apoptosis and Bcl-2 and Bax expression. RESULTS： Hypoxia and reoxygenation increased apoptosis over time, with Bcl-2 expression peaking at 24 hours and decreasing gradually （P 〈 0.01 ）; Bax expression peaked at 72 hours （P 〈 0.01）; the ratio of Bcl-2/Bax was 1.4, 0.8, and 0.6, respectively, at 24, 48 and 72 hours. Non-apoptotic astrocytes showed significant proliferation and swelling. Propofol treatment decreased apoptosis after hypoxia-reoxygenation （P 〈 0.01）, as well as Bct-2 and Bax expression （P 〈 0.05, P 〈 0.01）, with Bcl-2/Bax ratios of 1.6-1.8. Propofol treatmentalso blocked astrocyte proliferation and swelling. No apoptotic cells or Bcl-2/Bax expression was detected in astrocytes cultured in Hank＇s or Intralipid solution. CONCLUSION： Propofol protects astrocytes against injury caused by hypoxia and reoxygenation via a mechanism that involves maintaining high ratios of Bcl-2/Bax.

Preparation and Preservation of Hypoxia UW Solution

In order to explore the method to prepare hypoxia UW solution and the stability and preservation of hypoxia UW solution, UW solution was purged by argon or air for 15 min or 60 at a flow rate of 0.8 or 2 L/min, and the oxygen partial pressure of UW solution was detected. The hypoxia UW solution was exposed to the air or sealed up to preserve by using different methods, and the changes of oxygen partial pressure was tested. The results showed that oxygen partial pressure of 50 mL UW solution, purged by argon for 15 min at a flow rate of 2 L/min, was declined from 242±6 mmHg to 83±10 mmHg. After exposure to the air, oxygen partial pressure of hypoxia UW solution was gradually increased to 160±7 mmHg at 48 h. After sealed up by the centrifuge tube and plastic bad filled with argon, oxygen partial pressure of hypoxia UW solution was stable, about 88±13 mmHg at 72 h. It was concluded that oxygen of UW solution could be purged by argon efficiently. Sealed up by the centrifuge tube and plastic bag filled with argon, oxygen partial pressure of UW solution could be stabilized.

Extracorporeal membrane oxygenation for lung cancer-related life-threatening hypoxia:A case report

BACKGROUND Life-threatening hypoxia can occur in patients with lung cancer due to bronchial obstruction.Extracorporeal membrane oxygenation(ECMO)can be used as a bridge therapy for patients with severe hypoxia not relieved by conventional mechanical treatment.However,the usefulness of chemotherapy in patients with lung cancer receiving ECMO therapy is not well known.CASE SUMMARY A 53-year-old man visited the emergency room with worsening dyspnea for 1 mo.A series of imaging and diagnostic tests were performed,and stageⅢB(cT4N2M0)lung cancer was eventually diagnosed.On hospital day 3,he experienced dyspnea and hypoxia that was not relieved with oxygen support via a high-flow nasal cannula.ECMO was initiated because his respiratory condition did not improve even with mechanical ventilation.The patient then underwent gemcitabine/cisplatin chemotherapy without dose reduction while on ECMO.After two cycles of chemotherapy,there was a decrease in the size of the primary tumor in the right main bronchus.After the completion of concurrent chemoradiotherapy,a computed tomography scan revealed further improvement in the right main bronchus narrowing.Eight months after a lung cancer diagnosis,the patient did well without any dyspnea.CONCLUSION ECMO is a potential bridge therapy for respiratory failure in patients with central airway obstruction secondary to lung cancer.

Effect of hypoxia/reoxygenation on the viability expression of ROS and MAPKs in myocardial cells

Objective:To investigate to the expression effect of hypoxia and hypoxia/reoxygenation on ROS,MAPKs and cell apoptosis in H9c2 cardiomyocytes.Methods:H9c2 cells were treated with cobalt chloride(CoCl2)to establish the chemical hypoxia and hypoxia/reoxygenation-induced cardiomyocyte injury model.CoCl2 was used to process cells at different concentrations from 150-2400μmol/L and different time from4-24 h;H9c2 cells viability was detected by MTT,and the intracellular ROS level was measured by 2’,7’-dichlorflμoresceindiacetate(DCFH-DA)and dihydroethidiμm(DHE)staining and photoflurography.The active expression level of mitogen-activated protein kinases(MAPKs)(including JNK,ERK and p38)and caspase-3.Results:At the concentration from 300 to 1200μmol/L,CoCl2 does/time-dependently inhibited the cell viability in H9c2 cells(P<0.01).Compared with control group,the ROS levels in hypoxia group were significantly increased(P<0.05).In hypoxia group,the active expression levels of p-JNK,p-p38 and caspase-3 was higher than those in control group(P<0.05).However,the expression of p-ERK wasn’t significant differernce.Furthermore,all the expression levels of ROS,p-JNK,p-ERK,p-p38 and caspase-3 in H/R group were significantly raised compared with hypoxia group(P<0.01).Conclusions:Reoxygenation further aggravate chemical hypoxia induced cardiomyocyte oxidative stress injury by activating ROS/MAPKs signals,suggesting the role of myocardial ischemia/reperfusion injury in the pathogenesis of ischemic heart disease.

Fuzzy Logic for Solving an Optimal Control Problem of Hypoxemic Hypoxia Tissue Blood Carbon Dioxide Exchange during Physical Activity

This paper aims at using of an approach integrating the fuzzy logic strategy for hypoxemic hypoxia tissue blood carbon dioxide human optimal control problem. To test the efficiency of this strategy, the authors propose a numerical comparison with the direct method by taking the values of determinant parameters of cardiovascular-respiratory system for a 30 years old woman in jogging as her regular physical activity. The results are in good agreement with experimental data.

Expression of hypoxia-inducible factor 1 alpha and oligodendrocyte lineage gene-1 in cultured brain slices after oxygen-glucose deprivation

Oligodendrocyte lineage gene-1 expressed in oligodendrocytes may trigger the repair of neuronal myelin impairment, and play a crucial role in myelin repair. Hypoxia-inducible factor la, a transcription factor, is of great significance in premature infants with hypoxic-ischemic brain damage There is little evidence of direct regulatory effects of hypoxia-inducible factor le on oligodendrocyte lineage gene-l. In this study, brain slices of Sprague-Dawley rats were cultured and subjected to oxygen-glucose deprivation. Then, slices were transfected with hypoxia-inducible factor la or oligodendrocyte lineage gene-1. The expression levels of hypoxia-inducible factor la and oligodendrocyte lineage gene-1 were significantly up-regulated in rat brains prior to transfection, as detected by immunohistochemical staining. Eight hours after transfection of slices with hypoxia-inducible factor la, oligodendrocyte lineage gene-1 expression was upregulated, and reached a peak 24 hours after transfection. Oligodendrocyte lineage gene-1 transfection induced no significant differences in hypoxia-inducible factor la levels in rat brain tissues with oxygen-glucose deprivation. These experimental findings indicate that hypoxia-inducible factor la can regulate oligodendrocyte lineage gene-1 expression in hypoxic brain tissue, thus repairing the neural impairment.

Effects of astragalus polysaccharide on cell injury and mitochondrial pathway apoptosis in the hypoxia reoxygenation of myocardial cells

Objective: To study the effects of astragalus polysaccharide on cell injury and mitochondrial pathway apoptosis in the hypoxia reoxygenation of myocardial cells. Methods: H9c2 myocardial cell lines were cultured and divided into negative control group (NC group), hypoxia reoxygenation group (H/R group) and astragalus polysaccharide group (APS), H/R group underwent hypoxia reoxygenation treatment, and APS group underwent both hypoxia reoxygenation treatment and astragalus polysaccharides intervention. The cell viability was measured 8 h, 16 h and 24 h after reoxygenation;the expression of mitochondrial apoptosis genes, apoptosis pathway genes as well as the contents of ROS metabolism indexes were determined 24 h after reoxygenation. Results: 8 h, 16 h and 24 h after reoxygenation, the cell viability of H/R group were lower than those of NC group, and the cell viability of APS group were higher than those of H/R group;24 h after reoxygenation, BIM, BAX, Caspase-9, Caspase-3, p-PI3K, p-AKT and p-FoxO3a protein expression as well as ROS, HSP70 and p-p38MAPK contents in H/R group were significantly higher than those in NC group whereas BCL2 protein expression and SOD content were significantly lower than those in NC group;BIM, BAX, Caspase-9, Caspase-3, p-PI3K, p-AKT and p-FoxO3a protein expression as well as ROS, HSP70 and p-p38MAPK contents in APS group were significantly lower than those in H/R group whereas BCL2 protein expression and SOD content were significantly higher than those in H/R group. Conclusion: Astragalus polysaccharide can reduce the cell damage and inhibit the mitochondrial pathway apoptosis in the hypoxia reoxygenation process of myocardial cells.

Nano oxygen chamber by cascade reaction for hypoxia mitigation and reactive oxygen species scavenging in wound healing

Hypoxia,excessive reactive oxygen species(ROS),and impaired angiogenesis are prominent obstacles to wound healing following trauma and surgical procedures,often leading to the development of keloids and hypertrophic scars.To address these challenges,a novel approach has been proposed,involving the development of a cascade enzymatic reaction-based nanocarriers-laden wound dressing.This advanced technology incorporates superoxide dismutase modified oxygen nanobubbles and catalase modified oxygen nanobubbles within an alginate hydrogel matrix.The oxygen nano chamber functions through a cascade reaction between superoxide dismutase and catalase,wherein excessive superoxide in the wound environment is enzymatically decomposed into hydrogen peroxide,and this hydrogen peroxide is subsequently converted into oxygen by catalase.This enzymatic cascade effectively controls wound inflammation and hypoxia,mitigating the risk of keloid formation.Concurrently,the oxygen nanobubbles release oxygen continuously,thus providing a sustained supply of oxygen to the wound site.The oxygen release from this dynamic system stimulates fibroblast proliferation,fosters the formation of new blood vessels,and contributes to the overall wound healing process.In the rat full-thickness wound model,the cascade reaction-based nano oxygen chamber displayed a notable capacity to expedite wound healing without scarring.Furthermore,in the pilot study of porcine full-thickness wound healing,a notable acceleration of tissue repair was observed in the conceived cascade reaction-based gel treated group within the 3 days post-surgery,which represents the proliferation stage of healing process.These achievements hold significant importance in ensuring the complete functional recovery of tissues,thereby highlighting its potential as a promising approach for enhancing wound healing outcomes.