Progress on the relationship of aldehyde dehydrogenase 2 with human diseases and its small-molecule activators / 药学实践杂志

Aldehyde dehydrogenase 2 (ALDH2) is one of important factors against from the damage under oxidative stress in human body. A high proportion of East Asians carry ALDH2 inactive mutation gene. There are many diseases closely related to ALDH2, such as cardiovascular diseases, neurodegenerative diseases and liver diseases. Recent studies also have found that ALDH2 is associated with ferroptosis. Therefore, ALDH2 has becoming a potential target for the treatment of the above related diseases. Several types of small molecule activators with potential value of clinical application have been reported. The research progress on the structure and function of ALDH2 , the relationship with human diseases and its activators were summarized in this paper.

Investigation of acupuncture in improving sleep, cognitive and emotion based on attenuation of oxidative stress in prefrontal cortex in sleep-deprived rats / 针灸推拿医学（英文版）

Objective: To explore whether acupuncture can improve sleep disturbance, cognitive impairment and emotional disorders caused by sleep deprivation, and its association with the attenuation of oxidative stress injury in prefrontal cortex. Methods: Fifty-two male Sprague-Dawley rats were randomly divided into a control group (n=10), a model group (n=14), a manual acupuncture (MA) group (n=14), and a sham-MA group (n=14). All the groups were established as sleep deprivation models via the modified multiple platform method, except for the control group. Rats in both the MA group and the sham-MA group received corresponding intervention, respectively. After modeling and intervention, the four groups received three behavioral tests, namely sleep monitoring, by comprehensive lab animal monitoring system (CLAMS), Morris water maze (MWM) test and open-field test (OFT), followed by oxygen free radical level test and Western blot (WB) detection for the expression levels of Bax and Bcl-2. Results: The MA group derived more sleep time within 24 h than either the model group or the sham-MA group (both P<0.05). On MWM orientation navigation test day 1, there were no significant differences in escape latency among the control, MA and sham-MA groups (P>0.05), and the escape latency was significantly shorter in these three groups than that in the model group (all P<0.05). On test day 4, the escape latency was markedly shorter in the MA group than that in either the model group or the sham-MA group (both P<0.05); meanwhile, the MA group showed significantly better performance compared with these two groups in space probe test (both P<0.05). In OFT, compared with the control group, there was a significant decline in the horizontal movement score in the other three groups (all P<0.05), and the decrease was more significant in the model group and the sham-MA group than that in the MA group (both P<0.05). The superoxide dismutase (SOD) content was markedly higher and the malondialdehyde (MDA) content was markedly lower in the MA group than those in the model group and the sham-MA group (all P<0.05). Compared with the model group and the sham-MA group, the expression of Bax was significantly lower and the expression of Bcl-2 was significantly higher in the MA group (all P<0.05). Conclusion: MA therapy can lengthen the sleep time in sleep-deprived rats and improve learning and memory impairments induced by sleep deprivation, and the underlying mechanism may be associated with the enhancement of antioxidant capacity in the prefrontal cortex and the inhibition of hippocampal neuronal apoptosis.

Protective effect of extracorporeal membrane pulmonary oxygenation combined with cardiopulmonary resuscitation on post-resuscitation lung injury / 世界急诊医学杂志（英文）

@#BACKGROUND: Cardiac arrest (CA) is a critical condition that is a concern to healthcare workers. Comparative studies on extracorporeal cardiopulmonary resuscitation (ECPR) and conventional cardiopulmonary resuscitation (CCPR) technologies have shown that ECPR is superior to CCPR. However, there is a lack of studies that compare the protective effects of these two resuscitative methods on organs. Therefore, we aim to perform experiments in swine models of ventricular fibrillation-induced CA to study whether the early application of ECPR has advantages over CCPR in the lung injury and to explore the protective mechanism of ECPR on the post-resuscitation pulmonary injury. METHODS: Sixteen male swine were randomized to CCPR (CCPR; n=8; CCPR alone) and ECPR (ECPR; n=8; extracorporeal membrane oxygenation with CCPR) groups, with the restoration of spontaneous circulation at 6 hours as an endpoint. RESULTS: For the two groups, the survival rates between the two groups were not statistically significant (P>0.05), the blood and lung biomarkers were statistically significant (P<0.05), and the extravascular lung water and pulmonary vascular permeability index were statistically significant (P<0.01). Compared with the ECPR group, electron microscopy revealed mostly vacuolated intracellular alveolar type II lamellar bodies and a fuzzy lamellar structure with widening and blurring of the blood-gas barrier in the CCPR group. CONCLUSIONS: ECPR may have pulmonary protective effects, possibly related to the regulation of alveolar surface-active proteins and mitigated oxidative stress response post-resuscitation.

Regulation of Buyang Huanwu Decoction extracts on apoptosis and autophagy of PC12 cells model with oxidative stress injury / 中草药

Objective: To explore the underlying mechanism of Buyang Huanwu Decoction extracts on apoptosis and autophagy in PC12 cells model with oxidative stress injury. Methods: Different level oxidative stress injury models with H2O2 at various concentrations were established. The effective concentrations of Buyang Huanwu Decoction extracts were determined by MTT method in the initial stage and the intensifying period after oxidative stress injury. The apoptosis of PC12 cells were evaluated by FCM and TUNNEL, the autophagy situations were observed by TEM and mRFP-GFP-LC3. Furthermore, the proteins of Bax, Bcl-2, Beclin1, LC3A, and LC3B related to apoptosis were determined by Western blotting. Results: The initial stage and the intensifying period of oxidative stress injury cell models were established by H2O2 at the concentration of 1.5 and 2.0 mmol/L, respectively. Compared with the control group, model group appeared increasing apoptosis and autophagy levels, and model group had higher expressions of Bax/Bcl-2, Beclin1, LC3B and lower expression of LC3A (P < 0.05). Compared with the initial stage of oxidative stress injury cell models, Buyang Huanwu Decoction extracts could reduce the Bax/Bcl-2 and restrain apoptosis rates, while the autophagy was activated by up-regulation Beclin1 and LC3B/LC3A (P < 0.05). When the serious apoptosis and excessive autophagy were observed in the intensifying period of oxidative stress injury cells, the extracts could play the protective effect by apoptosis restraining and autophagy alleviating. Conclusion: Buyang Huanwu Decoction extracts can play the protective effects on oxidative stress injury cell models in different period by regulating apoptosis and autophagy.

Protective mechanism of hydrogen on oxidative stress-induced retinal aging / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To investigate the protective mechanism of hydrogen on retinal senescence induced by oxidative stress.<p>METHODS: Mice were randomly divided into three groups: control group, model group and treatment group. Animal models of retinal oxidative stress injury were established by injecting sodium iodate solution into mice caudal vein. Harvesting the mice retina, Western-blot was used to detect the level of proteins related to DNA damage response such as ATM, NF-κB, cyclin D1 and HMGB1 that associated with DNA repair.<p>RESULTS: SA-β-gal staining showed that the blue-green deposits in treatment group were reduced than that in model group. The expression of DNA damage reactive protein in treatment group ATM, cyclin D1, NF-κB(0.10±0.009, 0.32±0.01, 0.19±0.002)were significantly lower than those in the model groups(0.77±0.08, 0.70±0.02, 0.36±0.01), and the differences were statistically significant(all <i>P</i><0.01). At the same time, the expression of DNA repair protein HMGB1 in treatment group(0.927±0.06)were notably higher than that in model group(0.383±0.07)and the difference was statistically significant(<i>P</i><0.01).<p>CONCLUSION: H₂ can attenuate senescence by inhibiting oxidative-stress induced DNA damage.

Role of ERK signaling pathway in dexmedetomidine against mouse neuroblastoma N2a cell injury induced by oxidative stress / 临床麻醉学杂志

Objective To investigate the role and underlying mechanism of dexmedetomidine in protecting mouse neuroblastoma N2a cells against oxidative stress injury,and to discuss the effect of ERK signaling pathway.Methods Na2 cell oxidative stress injury model was established by H2O2 treatment.Cells were divided into 5 groups:control group (group C),H2O2group (group H), dexmedetomidine group (group D),H2O2+dexmedetomidine group (group HD),H2O2+dexme-detomidine+ERK inhibitor group (group HDP).Group H,group HD and group HDP were given 200 μmol/L H2O2with or without 100 ng/ml dexmedetomidine and 20 μmol/L ERK inhibitor PD98059,group D was treated with dexmedetomidine at the corresponding point,group C was treated with equal normal saline,After 1,4 hours of H2O2stimulation,cell survival,morphology changes,SOD production and ERK intracellular signaling pathway were compared between groups. Results Compared to group C,N2a cells in the group H demonstrated significantly ruduced cell sur-vival,much worse cell morphology and less SOD production (P<0.05).Compared to group H,N2a cells in group HD demonstrated significantly increased cell survival,much better preserved cell mor-phology,higher levels of SOD and enhanced ERK activation (P<0.05);Compared to group HD, cells in the group HDP had markedly decreased cell survival,worse cell morphology and lower SOD level (P<0.05).No significant changes were found in cell survival,morphology changes,SOD pro-duction and ERK intracellular signaling pathway between the groups C and D.Conclusion Dexme-detomidine protected mouse neuroblastoma N2a cells against oxidative stress injury by regulating ERK activation and SOD production.

Resveratrol protects human sperm against cryopreservation-induced injury / 中华男科学杂志

<p><b>Objective</b>To investigate the effects of resveratrol in the cryopreservation medium on the quality and function of post-thaw sperm.</p><p><b>METHODS</b>Semen samples were obtained from 50 normozoospermic and 50 oligoasthenozoospermic men, liquefied and then cryopreserved in the glycerol-egg yolk-citrate (GEYC) medium with or without 30 μmol/L resveratrol. Sperm motility, viability and acrosome reaction (AR) were examined before and after thawing. Sperm lipid peroxidation and the level of reactive oxygen species (ROS) were measured using commercial malondialdehyde (MDA) and the ROS assay kit. Sperm mitochondrial membrane potential (MMP) and DNA damage were determined by Rhodamine 123 staining and TUNEL.</p><p><b>RESULTS</b>The percentage of progressively motile sperm (PMS), total sperm motility, sperm viability, MMP and AR were significantly decreased (P <0.05) while the levels of sperm ROS, MDA and DNA fragmentation index (DFI) remarkably increased in both the normozoospermia and oligoasthenozoospermia groups after cryopreservation as compared with those in the fresh ejaculate (P <0.05). In comparison with the non-resveratrol control, the post-thaw sperm cryopreserved with 30 μmol/L resveratrol showed markedly higher PMS (［32.7 ± 4.8］ vs ［43.1 ± 6.3］ %, P <0.05), total motility (［44.8 ± 6.9］ vs ［56.9 ± 7.4］ %, P <0.05), viability (［52.3 ± 6.1］ vs ［67.5 ± 5.6］ %, P <0.05), MMP (［56.5 ± 7.0］ vs ［63.4 ± 7.5］ %, P <0.05) and AR (［16.6 ± 3.8］ vs ［26.3 ± 4.7］ %, P <0.05) but lower ROS, MDA and DFI (all P <0.05) in the normozoospermia group, and so did the post-thaw sperm in the oligoasthenozoospermia group, with a particularly lower DFI (［28.5 ± 4.8］ vs ［36.3 ± 5.7］%, P <0.01).</p><p><b>CONCLUSIONS</b>Resveratrol in the cryopreservation medium can improve the quality and function of post-thaw human sperm by reducing cryopreservation-induced sperm injury and the level of ROS.</p>

Protective effect of Polygonum orientale flower extract on H₂O₂-induced oxidative damage of HUVEC cells / 中国中药杂志

To investigate the protective effects and mechanism of Polygonum orientale flower extract on H₂O₂-induced oxidative damage of human umbilical vein endothelial cells (HUVEC), H₂O₂ was used to induce the oxidativestress damage on HUVEC cells and efforts were made to screen the low, medium and high drug concentrations of P.orientale flower extract. Cell viability was detected by the MTS assay. The content of lactate dehydrogenase (LDH) and malondialdehyde (MDA), and the activities of superoxidedimutase (SOD) and catalase (CAT) were detected by biochemical kits. The mRNA and protein levels of Bax, Bcl-2 were detected respectively by quantitative real time polymerase chain reaction (qRT-PCR) and Western blot. The protein level of cleaved caspase-3 was detected by Western blot. According to the results, the viability of HUVEC cells was reduced to around 55% after being treated with 120 μmol·L⁻¹ H₂O₂ for 0.5 h. Treatment of H₂O₂ also could increase LDH leakage rate and MDA content and attenuate the activities of SOD and CAT, up-regulate the expression level of Bax and cleaved caspase-3, and down-regulate the expression level of Bcl-2. As compared with H₂O₂ model group, P.orientale flower extract of 50-200 mg·L⁻¹ could increase the viability of HUVEC cells, reduce LDH release and MDA content, enhance the activities of SOD and CAT, down-regulate pro-apoptotic protein cleaved caspase-3 and Bax, and up-regulate apoptosis inhibitory protein Bcl-2. In summary, P.orientale flower extract showed a protective effect on H₂O₂-induced HUVEC cells injury, which may result from enhancing the cell capability of clearing the oxygen free radial, decreasing the production of lipid peroxidation and inhibiting apoptosis.

Role of mitochondrial permeability transition pore in mediating the inhibitory effect of gastrodin on oxidative stress in cardiac myocytes / 南方医科大学学报

OBJECTIVE@#To explore the role of mitochondrial permeability transition pore (mPTP) in mediating the protective effect of gastrodin against oxidative stress damage in H9c2 cardiac myocytes.@*METHODS@#H9c2 cardiac myocytes were treated with HO, gastrodin, gastrodin+HO, cyclosporin A (CsA), or CsA+gas+HO group. MTT assay was used to detect the survival ratio of H9c2 cells, and flow cytometry with Annexin V-FITC/PI double staining was used to analyze the early apoptosis rate after the treatments. The concentration of ATP and level of reactive oxygen species (ROS) in the cells were detected using commercial kits. The mitochondrial membrane potential of the cells was detected with laser confocal microscopy. The expression of cytochrome C was detected with Western blotting, and the activity of caspase-3 was also assessed in the cells.@*RESULTS@#Gastrodin pretreatment could prevent oxidative stress-induced reduction of mitochondrial membrane potential, and this effect was inhibited by the application of CsA. Gastrodin significantly lowered the levels of ROS and apoptosis-related factors in HO-exposed cells, and such effects were reversed by CsA. CsA significantly antagonized the protective effect of gastrodin against apoptosis in HO-exposed cells.@*CONCLUSIONS@#Gastrodin prevents oxidative stress-induced injury in H9c2 cells by inhibiting mPTP opening to reduce the cell apoptosis.

Protective effect of salvianolic acid B on cardiomyocytes of suckling rats with oxidative stress injury and its mechanism / 吉林大学学报(医学版)

Objective: To investigate the protective effect of salvianolic acid B on the cardiomyocytes of the suckling rats with oxidative stress injury, and to clarify its mechanism. Methods: The cardiomyocytes of Wistar suckling rats were primary cultured in vitro and randomly divided into normal control group, model group, low dose of salvianolic acid B group, and high dose of salvianolic acid B group. Except normal control group, the suckling rat cardiomyocytes were induced to oxidative stress injury by being exposed to H2O2 at the dose 100 μmoL · L-1; 20 and 40 μmol · L-1 salvianolic acid B were separatly added into low and high doses of salvianolic acid B groups. After co-culturing for 4 h, the morphological changes of cardiomyocytes were observed by light microscope; the survival rate of cardiomyocytes was examined using MTT. The levels of reductive glutathione (GSH), the activity of superoxide dismutase (SOD) and the level of malondialdehyde (MDA) in the cardiomyocytes were detected with spectrophotometry. The levels of lactate dehydrogenase (LDH) and creatine kinase (CK) in culture medium were determined by ELISA. Results: Compared with normal control group, the cardiomyocytes in model group partially suspended in the culture medium, the number of adherent cells was decreased, the pseudopod contraction and the volume were decreased, and the self-discipline throb of cardiomyocytes was slow. Compared with model group, the number of cardiomyocytes in 20 and 40 μmol · L-1 salvianolic acid B groups were increased significantly, the pseudopod contraction was reduced, and self-discipline throb was enhanced. Compared with normal control group, the survival rate of cardiomyocytes in model group was decreased (P<0. 01), the level of GSH in cardiomyocytes was decreased (P<0. 01), the level of MDA was increased (P<0. 01), the activity of SOD was decreased (P< 0. 01), and the levels of CK and LDH in culture medium were increased (P<0. 05 or P<0. 01). Compared with model group, the survival rates of cardiomyocytes in 20 and 40 μmol · L-1 salvianolic acid B groups were increased (P<0. 05 or P<0. 01), the levels of GSH and the activities of SOD were increased (P<0. 05 or P<0. 01), the levels of MDA were decreased (P<0. 05 or P<0. 01), and the CK and LDH levels in culture medium were decreased (P<0. 05 or P<0. 01). Conclusion; Salvianolic acid B can protect the cardiomyocytes against oxidative stress injury, and its mechanism may be associated with the inhibition of lipid peroxidatic reaction and the enhancement of antioxidant ability of cardiomyocytes.

The radiation protection role of heparin-SOD conjugate in irradiated mice

ABSTRACT Heparin-SOD conjugate (Hep-SOD) was prepared by modifying Cu,Zn-SOD with heparin. An acute radiation-induced mouse injury model was constructed to study the radiation protection effects of Hep-SOD conjugate. Fifty-six mice were randomly divided into seven groups: (I) normal control group; (II) irradiated control group; (III) positive control group (amifostine group, 300 mg/kg); (IV) SOD group (35000 U/kg); (V) high dosage of Hep-SOD group (70000 U/kg); (VI) medium dosage of Hep-SOD group (35000 U/kg); (VII) low dosage of Hep-SOD group (17500 U/kg). Drugs were intraperitoneally injected into each mouse 1 h before radiation except for the normal control group. All the irradiated groups were irradiated with 6 Gy. Organ indices, haematopoietic function indices, peripheral blood cells, liver function test, oxidative stress state and pathological observation were detected to study the effects of Hep-SOD on irradiated mice. Results showed that bone marrow suppression of irradiated mice could be reduced when treated by Hep-SOD before radiation. Oxidative stress detection and pathological observation of the liver and intestine showed that the damage caused by radiation was relieved when mice were treated with Hep-SOD before radiation. This study shows a new direction to prevent organisms from the damage caused by radiation.

Protective Effect of Blueberry Anthocyanins on Oxidative Stress Injury by H2O2 of Human Umbilical Vein Endothelial Cells / 医学研究杂志

Objective To study the protective effect of blueberry anthocyanins on H2O2 induced human umbilical vein endothelial cells,HUVECs.Methods The cultured cells were divided into 3 groups:normal control group,H2O2 model group,blueberry anthocyanins group.MTT method was used to observe cell viability.Annexin V-FITC/PI straining was used to observe cell apoptosis.Flow cytometry was used to detect cell cycle.Results H2O2could induce HUVECs oxidative stress injury.Cell survival rate was positive correlated with drug concentration and action time.In the certain concentration range blueberry anthocyanin had a protective effect.Conclusion Blueberry anthocyanin can protect H2O2induced oxidative stress injury in HUVECs.

Role of NADPH oxidase in high fat-induced oxidative stress injury in human umbilical vein endothelial cells / 中国药理学通报

Aim To explore the role of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase in high fat-induced oxidative stress injury in human umbilical vein endothelial cells (HUVECs).Methods HUVECs were exposed to different concentrations of palmitic acid(0.1,0.2,0.4,0.8 mmol · L-1) for 24 h and different time points of 0.4 mmol · L-1 palmitic acid(0,12,24,48 h).Cell viability was measured by Cell Counting Kit 8,and the protein expression of NADPH oxidase subunits such as p22phox,p47phox,p67phox and gp91phox were determined by Western blot.The expression of reactive oxygen species (ROS) in HUVECs was detected by immunofluorescence.Results Cell proliferation rate of HUVECs stimulated by 0.4 mmol · L-1 palmitic acid for 24 h and 48 h was significantly reduced.In the next experiment,model group was accordingly set as HUVECs stimulated by 0.4 mmol · L-1 palmitic acid for 24 h.The expression of NADPH oxidase subunits such as p22phox,p47phox,p67phox and gp91phox significantly increased at 24 h and 48 h after 0.4 mmol· L-1 palmitic acid stimulation (P ＜ 0.05),and the difference.between the 24 h group and the 48 h group was not significant (P ＞ 0.05).The expression of ROS in HUVECs significantly increased at 24 h and 48 h after O.4 mmol · L-1 palmitic acid stimulation (P ＜ 0.05),and the difference between 24 h group and 48 h group was not significant (P ＜ 0.05).Compared with the model group (0.4 mmol · L-1 palmitic acid stimulation for 24 h),the NADPH oxidase inhibitor diphenyliodonium (DPI,10 μmol · L-1) pretreatment could significantly decrease the expression of ROS in vascular endothelial cells (P ＜ 0.05).Conclusion Activated NADPH oxidase might play an important role in treatment of high fat-induced oxidative stress injury in vascular endothelial cells.

Effect of autophagy inhibitor chloroquine on the renal calcium oxalate crystals formation in rats / 中华泌尿外科杂志

Objective To investigate the effect and potential mechanism of autophagy inhibitor chloroquine on the calcium oxalate crystals formation in rats.Methods From September 2016 to October 2016,Thirty healthy male SD rats were randomly divided into 3 groups:control group,model group and chloroquine intervention group.The method to establish calcium oxalate stone model was drinking water with 1％ ethylene and 1％ ammonium chloride freely.The rats of chloroquine intervention group were treat with chloroquine (40mg/kg · d) by intraperitoneal injection.Modeling was finished after 28 days.The amounts of renalcalcium oxalate crystals were detected by polarizing microscope.For all groups,the amounts of autophagosome were detected by transmission electron microscope.Twenty four hour urine compositions for stone risk factors were detected.The expressions of oxidative stress injury related molecular markers (SOD,MCP-1 and 8-OHdG) and the expressions of autophagy markers (LC3 and P62) were detected by immunohistochemistry.The RNA expressions of SLC26A6 in kidney were detected by Real-time PCR.Results Compared to the model group,the amounts of renal calcium oxalate crystals were significantly reduced in chloroquine intervention group (32.37 ± 5.14 vs.4.18 ± 0.25,P ＜ 0.05).Compared to the control group,the level of autophagy was increased in the model group.Compared to the model group,the level of autophagy was inhibited in the chloroquine intervention group.For control group,model group and chloroquine intervention group,the excretion of urinary oxalate were (3.1 ± 1.5) mmol,(22.5 ± 8.1) mmol,(2.8 ± 1.2) mmol,respectively;the excretion of urinary citrate were (63.4 ± 7.4) mmol,(45.9 ± 9.5)mmol,(15.6 ± 8.2) mmol,respectively.Compared to the control group,the amounts of urinary oxalate weresignificantly elevated in model group (P ＜ 0.05),but citrate were significantly reduced in the chloroquineintervention group(P ＜ 0.05).For control group,model group and chloroquine intervention group,theexpressions of SOD were 42.24 ±4.16,19.21 ± 2.25,39.08 3.53,respectively;the expressions of MCP-1 were 4.02 0.51,8.45 ± 0.55,5.52 ± 0.34,respectively;the expressions of 8-OHdG were 7.16 ± 0.54,11.21 ± 1.12,8.67 ±0.34,respectively;the RNA expressions of SLC26A6 were 0.35 ±0.07,1.02 ±0.17,0.70 ± 0.06,respectively.Compared to the control group,the expressions of SOD were significantly reduced in the model group,but the expressions of MCP-1,8-OHdG and SLC26A6 were significantly elevated(P ＜0.05).Compared to the model group,the expressions of SOD were significantly elevated chloroquine intervention group (P ＜ 0.05),but the expressions of MCP-1,8-OHdG and SLC26A6 were significantly elevated(P ＜ 0.05).Conclusions The autophagy inhibitor chloroquine could inhibit the formation of calcium oxalate crystals induced by ethylene in rat kidney via inhibit the renal autophagy level and expressions of the SLC26A6,reducing the renal oxidative stress injury and urinary oxalate excretion.

Preincubation with low dose of hydrogen peroxide enhances anti-oxidative stress potential of mouse BMSCs / 基础医学与临床

Objective To investigate the effects of preconditioning with low-concentration hydrogen peroxide ( H2O2 ) on oxidative stress-induced bone marrow mesenchymal stem cells ( BMSCs ) apoptosis and its mecha-nism.Methods Mouse bone marrow mesenchymal stem cells ( BMSCs) were isolated and purified by differential centrifugation, and were treated with 0,200,250,300, 500 μmol/L H2O2 after preincubation with 50 μmol/L H2O2 or control medium.Apoptosis of these cells was measured by flow cytometry, and the expression of phos-phorylated PI3K, Akt and mTOR was analyzed by Western blot; BMSCs were also primed with PI3K inhibitor LY294002 for 30 min, then preincubated with 50 μmol/L H2O2 or control medium for 12 h before treatment with 300 μmol/L H2O2.Expression of apoptosis proteins Bcl-2, Bax, caspaase-3, cleaved-caspase-3 and the key pro-teins of the PI3K/Akt/mTOR pathway were detected by Western blot .Results H2O2 induced BMSCs apoptosis in a dose-dependent manner ,and pretreatment of BMSCs with low concentration of H2O2 significantly decreased H2O2-induced apoptosis of the BMSCs .Western blot results revealed that preconditioning with low-concentration H2O2 re-markably reversed the decrease in Bcl-2, total and phosphorylated PI3K, Akt and mTOR levels, and increased in Bax, cleaved-caspase-3 expression after high-dose H2O2 treatment.Such effects were antagonized by PI3K inhibitor LY294002 .Conclusions Preincubation with low-concentration H2O2 may indnce resistance of BMSC to oxidative stress, and such effect may be mediated by inhibition of pro-apoptotic proteins and activation of the PI 3K/Akt/mTOR pathway .

Protective effects and mechanism of extract from Cajanus cajan leaves against H2O2-induced oxidative injury on H9c2 cardiomyoblasts / 中草药

Objective: To investigate the protective effects and mechanism of the extract from Cajanus cajan leaves (ECCL) against H2O2-induced oxidative injury in H9c2 cardiomyoblasts. Methods: A model of H2O2-induced injury in H9c2 cardiomyoblasts was established. Cell viability was determined colorimetrically by MTT assay. The supernates and cells were collected, respectively, after the different treatments for measuring the LDH, MDA, and SOD levels with the corresponding detection kit according to the manufacturer's instructions. Western blotting was performed to exam the expression of p-Akt and p-eNOS in H9c2 cells respectively. Results: Compared with H2O2 group, the cell viability was increased significantly in ECCL + H2O2 groups (P < 0.01). The activity of LDH in the culture medium was decreased significantly (P < 0.05). The content of MDA in the culture medium was decreased significantly (P < 0.05). The activity of SOD was increased significantly (P < 0.01). Treated with ECCL, the expressions of p-Akt and p-eNOS in H9c2 cells injured from H2O2 were increased significantly (P < 0.01), When LY294002 (inhibitor of PI3K) was added, the effects of ECCL were cancelled. Conclusion: ECCL protects H9c2 cells against H2O2-induced oxidative injury partly through PI3K signaling pathway.

MicroRNA expression in hepatocytes with hydrogen peroxide-induced oxidative stress-injury and alleviating effect of mesenchymal stem cell-conditioned medium / 中国病理生理杂志

AIM: To evaluate the changes of microRNA (miRNA) in hepatocytes during hydrogen peroxide-induced oxidative stress injury, and to observe the alleviating effect of mesenchymal stem cell-conditioned medium (MSC-CM) in this progress.METHODS: The hepatocyte oxidative stress injury model was established using hydrogen peroxide and human normal liver cell line L02.MSC-CM was prepared using centrifugation and filter.The effects of MSC-CM on hepatocyte injury were evaluated by apoptosis analysis, cell viability detection, cell cycle, and mitochondrial membrane po-tential (MMP).Twenty-one differentially expressed miRNAs were selected by gene chip hybridization, in which miR-143, miR-145, miR-301a and let-7a were confirmed by RT-qPCR.Bioinformatics software was utilized to predict target proteins of these miRNAs, and then the proteins were verified by Western blot.RESULTS: MSC-CM markedly attenuated hydrogen peroxide-induced oxidative stress injury by reducing apoptosis, promoting cell viability and regulating cell cycle.The ex-pression of miR-143, miR-145, miR-301a and let-7a, indentified by RT-qPCR, increased under the condition of oxidative stress injury, while decreased after MSC-CM treatment.The expression of miR-143 predicted target proteins, HK2 and ADRB1, decreased under the hydrogen peroxide-exposure, while increased after MSC-CM treatment, which is consistent with the regulatory trend of miR-143.CONCLUSION: MSC-CM might attenuate hydrogen peroxide induced oxidative stress injury via inhibiting apoptosis and regulating some miRNA expression.

Effects of Postaglandin E1 on the Oxidative Stress Injury Index and the Expression of PCNA in Vein Grafts of Rabbit Carotid Arteries

BACKGROUND: This study investigated the protective mechanism of Prostagladin E1 (PGE1) against intimal hyperplasia after vein interposition grafts in rabbits. It has been demonstrated that active oxygen species contribute to vascular smooth muscle cell growth via early cell cycle gene activation. We attempted to study whether PGE1 had an effect on the inhibition of the oxidative stress injury index (8-OHdG, MDA). METHODS: Forty-eight jugular vein grafts were inserted into the carotid arteries of male hyperlipidemic New Zealand white rabbits, which were divided into 2 groups (saline group and PGE1 group). Saline and Prostaglandin E1 (0.1 microgram/kg/min) were administered as a continuous infusion for 2 hours every day from just before graft interposition to harvest. The vein grafts were harvested at 6 hour, 1 day, 1 week, and 2 week after grafting and rapidly stored in liquid nitrogen ( 70oC). 8-OHdG was measured by using high performance liquid chromatography coupled with electrochemical detection (HPLC-EC), and malondialdehyde (MDA) was measured by using thiobarbituric acid (TBA) assay. PC 10 index and intimal thickness of the grafts were measured with a computer digitalized image analyzer. RESULTS: There was no difference in 8-OHdG levels between the saline and the PGE1 groups. PGE1 had more inhibitory effect on the MDA level as an oxidative stress injury index, but its action was restricted to 1 day. A morphometric analysis and an immunohistochemical study showed that the PGE1 group had more suppressive effects both in intimal thickeness and proliferating cell nuclear antigen (PCNA) expression than the saline group (p<0.05). CONCLUSION: These results suggest that PGE1 is effective in preventing intimal hyperplasia after vein interposition grafts in rabbits and may play a role in inhibiting oxidative stress injury.

Expression of PCNA and Oxidative Stress Injury with Intimal Hyperplasia in Rabbit Experimental Vein Graft

Proliferation of vascular smooth muscle cells (VSMCs) is the utmost important pathophysiologic mechanism of intimal hyperplasia and atherosclerosis. With a hyperlipidemic rabbit model of jugular vein graft to carotid artery, we invesigated the oxidative stress injury in intimal hyperplasia and correlation of PCNA expression with VSMCs proliferation and intimal hyperplasia. Twenty jugular vein grafts were inserted into the carotid arteries of male New Zealand White rabbit and the vein grafts were harvested at 6 hr, 1 day, 7 days, 14 days, respectively after grafting and rapidly stored in buffered formalin for morphometric analysis, PCNA expression or frozen in liquid nitrogen for MDA (Malondialdehyde) analysis. Total wall and intimal thickness of grafts were measured with an computer digitalized image analyzer. Intimal thickening was rapidly increased at 7 days and peaked at 14 days (125.05 19.80 and 180.25 6.38 mum, respectively) and significantly thicker than control group or 6 hr, 1day after graft implantation (p<0.05). MDA level was significantly higher in vein grafting groups than control group (9.13 1.80 vs. 6.08 1.00 muM/mg protein, p=0.011) by Ohkawa method. In immunohistochemical staining, expression for PCNA (PC10 index: %) in media was peaked at 7 days (12.91 1.22) and significantly higher than control group or 6 hr, 1 day (0, 0.66 0.90, and 3.00 1.22, respectively) after graft implantation (p<0.05) and decreased thereafter. Expression for PCNA (PC10 index) in intima was markedly noted at 7 days (8.60 0.95), peaked at 14 days (16.90 2.14) and significantly higher than control group or 6 hr, 1 day (0, 0.31 0.63, and 1.44 1.00, respectively) after graft implantation (p<0.05). In conclusion, oxidative stress injury by active oxygen species increased in this model of vein graft suggests a role of this oxidant in intimal hyperplasia. PCNA expression was well correlated with the proliferation and migration of VSMCs from media to intima as the pathophysiology of intimal hyperplasia. Therefore, use of PCNA expression in this model of vein graft provides a reproducible method of assessing cellular proliferation after vascular injury. This experimental models of vessel wall injury are helpful in understanding the pathophysiologic mechanism of intimal hyperplasia and are useful in initial assessment of agents intended for reducing intimal hyperplasia.