Protective effect of puerarin against anoxia reoxygenation injury in cardiomyocytes based on VDAC1/mPTP regulation mechanism / 中国药理学通报

Aim To study the effect of puerarin( Pue) against myocardial ischemia/reperfusion injury and in-volved mitochondrial mechanism. Methods Anoxia/reoxygenation( A/R) injury model was established in H9c2 cell. Recombinant plasmid pFLAG-VDAC1 was constructed. Cells were randomly divided into 4 groups, normal control group ( Control) , A/R group, puerarin group ( Pue + A/R ) , and pFLAG-VDAC1-Pue group. Real-time PCR was used to investigate the expression of VDAC1 at mRNA level, and the expres-sion of protein level was detected by Western blot. LDH and CK activities were measured by automatic bi-ochemical analyzer. Mitochondrial membrane potential ( Δψm) and cell apoptosis were observed by flow cy-tometry method. Mitochondrial swelling test was used to detect the opening of mitochondria permeability tran- sition pore ( mPTP) . Results Compared with control group, the expression of VDAC1 and mRNA was up-regulated in A/R group, LDH and CK activity were el-evated, and then mPTP opened, Δψm collapsed, cell apoptosis was significantly increased. Puerarin pre-treatment can lower the expression of VDAC1, main-tain Δψm, prevent the opening of mPTP, and reduce apoptosis. However, the protective effect of Puerarin could be cancelled by transfection of pFLAG-VDAC1. Conclusions The cardioprotection of Puerarin against A/R injury is closely related to down-regulation of VDAC1 and prevention of mPTP opening.

Hydroxysafflor yellow A reduces anoxia/reoxygenation-induced injury in rat cardiomyocytes / 基础医学与临床

Objective To observe the protective effect of hydroxysafflor yellow A(HSYA) on anoxia/reoxygenation (A/R) injury of neonatal primary cardiomyocytes, and its relationship with phosphoinositide 3-kinase/protein ki-nase B/glycogen synthase kinase 3β(PI3K/Akt/GSK3β) signaling pathway. Methods Primary cardiomyocytes of neonatal rats were isolated from the rats and incubated for 48 hours. The cells were adhered to each other and then divided into five groups:control group (Con group), anoxia/reoxygenation group (A/R group),HSYA treatment group(A/R+H group),PI3K inhibitor (LY294002)treatment group(A/R+L group)and HSYA+LY294002 treat-ment group (A/R+H+L group),then to collect the supernatant fluid of each group to measure LDH.The flow cy-tometry was used to measure the apoptotic cells. The protein levels of Bcl-2,Bax,Akt,p-Akt (Ser473),GSK3β, p-GSK3β (Ser9) were evalated by Western blot. Results A/R increased LDH release,the apoptosis rate (P<0.001),and the expression of pro-apoptotic protein Bax (P <0.001) with the decrease of anti-apoptotic protein Bcl-2,p-Akt(Ser473), p-GSK3β(Ser9)(P<0.001) as compared with the control group. HSYA treatment de-creased LDH release,the apoptosis rate (P<0.001),and the expression of Bax (P<0.001) and increase the ex-pression of Bcl-2,p-Akt(Ser473),p-GSK3β(Ser9)(P<0.001). Compared with the A/R+H group,the expres-sion of Bax was increased (P<0.001),while the expression of Bcl-2, p-Akt(Ser473), p-GSK3β(Ser9)was de-creased (P<0.001) in the A/R+H+L group. Conclusions HSYA protects rats'cardiomyocytes from anoxia/reoxy-genation injury by regulating PI3K/Akt/GSK3β signaling pathway.

Inhibitory effect of Polygonatum sibiricum polysaccharides on release of inflammatory cytokines of anoxia/reoxygenation H9c2 myocardial cells through TLR4-MyD88-NF-κB signaling pathway / 中国药理学通报

Aim To assess the regulatory effects of Po-lygonatum sibiricum polysaccharides(PSP)on Toll-like receptor 4 (TLR4 )-myeloid differentiation factor 88 (MyD88)-nuclear factor κB(NF-κB)signaling path-way in anoxia /reoxygenation-H9c2 myocardial cells. Methods The H9c2 myocardial cells cultured in vitro were randomly divided into groups:control group (C group),hypoxia /reoxygenation group (H/R group), PSP group,TLR4 inhibitor group(TAK-242 group)and PSP +TLR4 inhibitor group(PSP +TAK-242 group). The cells were cultured in normal condition for 27 h in C group.The cells were subjected to 21 h hypoxia fol-lowed by 6 h reoxygenation in H/R.Definitely,the cells in TAK-242,PSP and PSP +TAK-242 groups were treated with PSP and TAK-242 with the final con-centration of 1 .5 g·L -1 and 1 μmol·L -1 for 1 2 h before 21 h hypoxia,then the cells were exposed to the normal culture condition for another 6 h.After the treatment,cell survival rate was tested by MTT method. The contents of tumor necrosis factor-α(TNF-α)and interleukin-1 β(IL-1 β)were determined by enzyme-linked immunosorbent assay(ELISA).The protein ex-pression levels of NF-κB and inhibitor κBα(IκBα) were detected by Western blot,and the expression lev-els of TLR4 and MyD88 mRNA were detected by fluo-rescence quantitative PCR method.Results Compared with H/R group,the cell survival rates were significant-ly increased,while the inflammatory cytokines contents and NF-κB protein expression were dramatically de-creased in groups PSP,TAK-242 and PSP +TAK-242, whereas the NF-κB expression was significantly down-regulated,and the IκBα protein expression was in-creased.The mRNA expression levels of TLR4 and MyD88 were markedly decreased.Conclusion PSP might protect H9c2 myocardial cells against H/R inju-ry,which may be associated with the inhibition of TLR4-MyD88-NF-κB pathway.

Isoflurane attenuates human cardiacmyocytes anoxia/reoxygenation injury / 基础医学与临床

Objective_To explore the effect and the mechanism of isoflurane on human cardiac myocytes ( HCM) injury induced by anoxia/reoxygenation ( AR) .Methods_HCM cells were divided into control group ( con) , an-oxia/reoxygenation group (AR) and isoflurane (0.5%, 1%, 1.5%and 2%) treatment group (n=6).Cell via-bility, LDH activity, apoptosis and the expression level of Anoxia inducible factor-1α( HIF-1α) were detected using CCK-8 assay, LDH activity assay kit, Annexin V-FITC/PI staining, PCR and western blot, respectively. Results_Compared with the con group, cell viability decreased, LDH activity and apoptosis cells increased in AR group.Isoflurane can significantly relieve the decrease of cell viability, the increase of LDH activity and apoptosis cells, and the down-regulation of the mRNA and protein expression level of HIF-1αinduced by AR ( P<0.05 ) . Compared with AR group, the mRNA and protein expression level of HIF-1αin siRNA transfected group signifi-cantly decreased (P<0.05).2%isoflurane significantly relieve the increase of cell viability, the decrease of LDH activity and apoptotic cells induced by HIF-siRNA in AR injuried HCM cells(P<0.05).Conclusions_Isoflurane can protect HCM cells from AR injury partly through up-regulate the expression of HIF-1α.

Role of RISK signal pathway in reducing clenbuterol-induced cardiomycytes A/R injury of neonatal rat / 中国药理学通报

Aims To study the effects of clenbuterol on anoxia/reoxygenation( A/R) injury in neonatal Wistar rat cardiomyocytes and to explore whether its mecha-nism is related to reperfusion injury salvage kinase ( RISK) or not. Methods The cultured primary neo-natal cardiomyocytes were randomly divided into eight groups: ①normal culture group; ②anoxia/reoxygen-ation( A/R) group;③ clenbuterol ( 1 μmol · L-1 ) +A/R;④ICI118,551(10 μmol·L-1) + clenbuterol ( 1 μmol · L-1 ) + A/R; ⑤Metoprolol ( 10μmol · L-1 ) + clenbuterol(1μmol·L-1 ) + A/R group;⑥Metoprolol ( 10 μmol · L-1 ) + A/R group; ⑦PD98059 ( 20 μmol · L-1 ) + clenbuterol ( 1 μmol · L-1 ) + A/R group;⑧ LY294002(10 μmol·L-1 ) +clenbuterol(1 μmol · L-1 ) + A/R group. Cell via-bility was determined by the conventional MTT reduc-tion assay. The content of LDH in cultured medium was measured with colorimetry. Cardiomyocyte apopto-sis was determined by Hoechst33342 . Intracellular re-active species( ROS) were monitored by the fluorescent DCFH-DA. Total ERK2 and phosphorylated ERK were detected by western blot. Results Compared with A/R group, clenbuterol significantly increased vaibility of cells, reduced LDH release, lowered the rate of apop-tosis and ROS production. When addedβ2 receptor an-tagonist ICI118 , 551 , PI3 K inhibitor LY294002 and ERK inhibitor PD98059 , the effects of clenbuterol a-bove were inhibited; but β1 receptor antagonist Meto-prolol protected the cardiomyocytes from A/R injury, as evidenced by decreased LDH release and increased cell viability. There were no synergistic effects in the combined use of clenbuterol and Metoprolol. Conclu-sion clenbuterol exerts cardioprotective effects against A/R injury by inhibiting oxidative stress and apopto-sis. The protection of clenbuterol is inhibited by ICI118 , 551 , LY294002 and PD98059 . clenbuterol protects cardiomyocytes against A/R injury via RISK pathway by activation of β2 receptor.

Role of Bcl-2 signal pathway in apigenin preconditioning against cardiomyocytes anoxia/reoxygenation injury / 中国药理学通报

Aim To investigate the relationship be-tween the cardioprotection of apigenin ( Api ) from an-oxia/reoxygenation ( A/R) injury and Bcl-2 pathway. Methods H9 c2 cardiomyocytes were cultured and di-vided into normal control group, A/R group, Api pre-treatment group ( Api ) , Api + Bcl-2 inhibitor group ( Api + ABT-737 ) . Expression of Bcl-2 was deter-mined by Western blot,and cell viability was measured by MTT method. LDH, SOD, GSH-Px, MDA activity were determined by chromometry. ROS generation, mi-tochondrial membrane potential and apoptosis were de-termined by flow cytometry. Results 25h after apige-nin precondition,the expression of Bcl-2 was upregulat-ed in cardiomyocytes ( P <0. 01 ) . In the group pre-treated with 40 μmol · L-1 apigenin before A/R, the activity of LDH in culture medium decreased; the ac-tivity of intracellular SOD, GSH-Px increased; the content of MDA and ROS generation decreased; cell viability increased; mitochondrial membrane potential could be more stable and cell apoptosis decreased ( P<0. 01 ) . However, all these protective effects were attenuated significantly in the group pretreated with apigenin and Bcl-2 inhibitor ABT-737 . Conclusion The effect of apigenin against A/R injury in cardiomyo-cytes involves Bcl-2 pathway, and at least partly de-pends on its effect on upregulating the expression of Bcl-2 .

Effect of emulsified isoflurane on apoptosis of anoxia-reoxygenation neonatal rat cardiomyocytes

OBJECTIVE@#To explore the effect of emulsified isoflurane (EI) on apoptosis of anoxia-reoxygenation neonatal rat cardiomyocytes and relevant protein expression.@*METHODS@#Cardiac muscle anoxia-reoxygenation damage model was established with culture in vitro neonatal rat cardiomyocytes. The cardiomyocytes were divided into control group, model group, fat emulsion group and EI group. The cardiomyocytes apoptosis rates and lactic dehydrogenase (LDH), superoxide dismutase (SOD) and malondialdehyde (MDA) index standardization were detected after relevant treatment. The expression of apoptosis-related proteins Bel-2, Bax and Caspase-3 were detected with Western blot approach.@*RESULTS@#After hypoxia/reoxygenation (H/R) model was treated by EI, the cells apoptosis rate decreased and was dramatically below the fat emulsion group (P<0.05). Cardiomyocytes biochemical index detection presented that, compared with the control group that the LDH activity and MDA content dramatically increased (P<0.05), while the SOD activity notably decreased (P<0.05); compared with the H/R group, the SOD activity of the fat emulsion group and EI group increased (P<0.05); while the LDH activity and MDA content decreased (P<0.05). And the change of the EI group was more remarkable than the fat emulsion group (P<0.05). The Western blot analysis presented that, compared with the control group, the Bcl-2 protein expression of the other groups significantly decreased (P<0.05), the expressions of Bax protein and Caspase-3 protein increased significantly (P<0.05); compared with H/R group, cardiomyocytes Bcl-2 protein expression of EI group increased significantly (P<0.05), the expressions of Bax protein and Caspase-3 protein decreased significantly (P<0.05), and the change of EI group was more remarkable than the fat emulsion group (P<0.05).@*CONCLUSIONS@#EI can inhabit the apoptosis of anoxia-reoxygenation damage model cardiomyocytes, and may be related to the up-regulation of expression of Bcl-2 and down-regulation of expression of Caspase-3 protein.

Protection of echinocystic acid on primary cultured rat cardiomyocytes subjected to anoxia/reoxygen-ation injury / 中国药学杂志

OBJECTIVE: To study the protection effects of echshinone acid (EA) on the primary cultured rat cardiomyocytes subjected to anoxia-reoxygenation (A/R) injury. METHODS: The primary cultured neonatal rat cardiomyocytes were pretreated with EA (0.5, 5 and 50 μmol · L-1), EA (5 μmol · L-1) and L-NAME (0.1 mmol · L-1), or PD98059 (50 μmol · L-1) respectively for 1 h, and then subjected to A/R injury after 24 h. The cell viability, activities of SOD and GSH-Px, MDA contents, LDH activity in the medium and HSP70 protein expression were measured. RESULTS: Pretreatment with Ech decreased LDH activity and MDA contents, increased cell viability and SOD and GSH-Px activities in a concentration-dependent manner, and increased HSP70 protein expression. The heart protective effects of EA were partly abolished by L-NAME or PD98059. CONCLUSION: Pretreatment with EA for 1h before ischemia can induce delayed cardiomyocyte protective effects by activation of NO and MAPK signaling pathways and increasing expression of HSP70 in rat neonatal cardiomyocytes.

Effect of calcium-sensing receptor on apoptosis of neonatal rat cardiomyocytes / 国际儿科学杂志

ObjectiveTo observe the influence of calcium - sensing receptor (CaSR) on Fas/Fas ligand (Fas L) pathway during anoxia/reoxygenation (A/R)- induced cardiomyocytes apoptosis in neonatal rat. MethodsSingle cells were dissociated from minced hearts of 2 - day - old Wistar rats with a 0. 25％ solution of crude trypsin and then cultured as monolayers at a density of 5 x 104cells/cm2 in DMEM medium equilibrated with humidified air containing 5％ CO2 at 37C. Three days after the cells were seeded, the cultured cardiomyocytes were randomly divided into three groups. ( 1 ) control group: cardiomyocytes were continuously cultured for 26 hours in DMEM medium.(2) A/R group: cardiomyocytes underwent anoxia for 2 hours and reoxygenation for 24 hours.(3) GdCl3 group: 300μmol/L GdCl3 was added to the culture medium at the beginning of reoxygenation. Apoptosis of cardiomyocytes was assessed by flow cytometer and morphological alterations were observed with transmission electron microscope.The expression of CaSR, Fas and Fas L were analyzed by Western blot.Results The result of flow cytometer showed that cardiomyocytes apoptosis was 12. 18％ ± 1.54％ in A/R group,and was higher than that in the control group (P ＜0. 01 ). At the same time, mitochondrial cristae dissolution and disappearance could be detected. Compared with A/R group, GdCl3, a specific activator of CaSR, further enhanced cardiomyocytes apoptosis to 20. 25％ + 2. 87％ ( P ＜ 0. 01 ), along with an increment in CaSR, Fas and Fas L expressions ( P ＜ 0. 05 ). ConclusionCaSR is closely involved in cardiomyocytes apoptosia during anoxia/reoxygenation. CaSR could induce apoptosis of neonatal rat cardiomyocytes through Fas/Fas L receptor pathway.

Action mechanism of hyperin on neonatal rat's neuron with anoxia-reoxygenation / 中国药理学通报

Aim To study the action mechanism of hyperin(Hyp) on neonatal rat's neuron with anoxia/reoxygenation(A/R).Methods The dissociated neonatal rat brain cells were subjected to 30 min of anoxia or followed 40 min of reoxygenation.Lactate dehydrogenase(LDH),malondialdehyde(MDA)and nitric oxide(NO)in the supernatant were measured.The intracellular free calcium concentration([Ca~(2+)]_i)in brain cells was assayed with Fura 2-AM method.Results Anoxia induced a significant increase of LDH in the supernatant from (62.0±13.0) U·L~(-1)(Sham group)to (116.0±16.6) U·L~(-1)(Control group,P<0.01),and reoxygenation markedly increased LDH and MDA in the supernatant from (45.6±9.2) U·L~(-1) and (9.1±0.9) μmol·L~(-1)(Sham group)to (106.0±17.4) U·L~(-1) and (16.4±2.7) μmol·L~(-1)(Control group,P<0.01),respectively.In the range of 1.0 ～ 16.0 μmol·L~(-1),Hyp markedly and concentration-dependently inhibited anoxia-or reoxygenation-evoked increases of LDH and MDA.1.0～16.0 μmol·L~(-1) Hyp not only inhibited anoxia-induced increase of NO in the supernatant and rise of [Ca~(2+)]_i in brain cells(P<0.05 or P<0.01),but also attenuated reoxygenation-evoked increases of NO and[Ca~(2+)]_i(P<0.05 or P<0.01),Hyp 16.0 μmol·L~(-1) significantly reduced NO and[Ca~(2+)]_i from (34.4±6.3) μmol·L~(-1) and (640±94) nmol·L~(-1) to (25.0±5.1) μmol·L~(-1) and (331±56) nmol·L~(-1),respectively.Conclusion The protective effect of Hyp on A/R-injured neurons may be related to the inhibition of overload of[Ca~(2+)]_i,NO release and lipid peroxidation.

Effect of calcium-sensing receptor on anoxia/reoxygenation-induced apoptosis in cultured cardiomyocyte of neonatal rats / 国际儿科学杂志

Objective Calcium-sensing receptor(CaSR)belongs to the family C of G-protein coupled receptors.This study was carried out to observe the influence and mechanism of CaSR on anoxia/reoxygenation(A/R)-induced cardiomyocytes apoptosis.Methods The model of A/R injury was established through anoxia for 2 hours and reoxygenation for 24 hours in cultured cardiomyocytes of neonatal rats.Cardiomyocytes were randomly divided into three groups:control group,A/R group and GdCl3 group(300μmol/L GdCl3 was added to the culture medium at the beginning of reoxygenation).Apoptosis of cardiomyocytes was assessed by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL).The expression of CaSR,cysteine-requiring aspartate protease(caspase)-3,9 and cytochrom c (Cytc)were analyzed by Western blot.Results The TUNEL showed that cardiomyocytes apoptosis was 17%±3% in A/R group,and was higher than that in the control group.At the same time,expression of CaSR in A/R group was markedly increased in response to control group.Compared with A/R group,GdCl3,a specific activator of CaSR,further enhanced cardiomyocytes apoptosis to (28±4)% and decreased the ability of cardiomyocytes to (51.2±6.8)%,along with an increment in CaSR,caspase-3,caspase-9 and Cytc expressions.Conclusion CaSR is involved in anoxia/reoxygenation-induced apoptosis of neonatal rat cardiomyocytes through Cytc-caspase-3 pathway.

Involvement of COX-2 in antagonism to heme oxygenase-1 on cardioprotection from anoxia/reoxygenation induced injury in rats / 基础医学与临床

Objective To investigate the role of HO-1 in the protection of rat heart from anoxia/reoxygenation induced injury and its underlying mechanism.Methods LVEDP,LVDP and dp/dtmax were analyzed by the Langendorff method in isolated rat heart.Lactate dehydrogenase(LDH), infarct area,COHb and 6-keto-PGF1? were further determined in the experiment.Results After intraperitoneal injection of HO-1 inducer hemin,CO concentration in rat blood enhanced(P

Effect of Huoxuetongluo Decoction on Expression of CD54 and Polymorphonuclear Adhesion during Anoxia/Reoxygenation Injury in Human Umbilical Vein Endothelial Cells / 中国中医药信息杂志

Objective To investigate the effect of Huoxuetongluo Decoction on expression of CD54 and polymorphonuclear adhesion during human umbilical vein endothelial cells(HUVEC) anoxia/reoxygenation injury,and explore its possible mechanisms.Methods Rabbits serum containing Huoxuetongluo Decoction was prepared.Resuscitate and culture HUVEC,then establish the anoxia/reoxygenation injury model of HUVEC.The model cells were divided into five groups:normal control,model group and group of high,medium,low dose of Huoxuetongluo Decoction.After dealt seperately,the morphologic change of cells were observed through the microscope,the expression of CD54 and the polymorphonuclear adhesion were determined.Results The group of low,medium and high dose of Huoxuetongluo Decoction inhibited the expression of CD54 and decreased the adhesion between polymorphonuclear and HUVEC,the effects were strengthened with increasing the dose of Huoxuetongluo Decoction.The difference between group of medium,high dose of Huoxuetongluo Decoction and model group had statistical significance(P

Effects of n-butanol Extract of Morindae officinalis How on Apoptosis of Myocardial Cells in Neonatal Rats Undergoing Anoxia/Reoxygenation / 中国药房

OBJECTIVE: To investigate the effect of n-butanol extract of Morindae officinalis How(BEM) on apoptosis of myocardial cells in neonatal rats undergoing anoxia/reoxygenation(A/R).METHODS: The rats were divided into 6 groups: normal group,A/R model,Danshen ingection(SM),BEM high,middle,low groups.The morphological changes of the apoptotic myocardial cells were observed using electron microscope.The apoptotic index was measured by in situ nick end labeling(TUNEL).The expression of Bcl-2 and the apoptotic protein Bax were detected by immunohistochemical staining.RESULTS: Morphocytology revealed the remarkable protective effect of BEM against apoptosis of myocardial cells during A/R.TUNEL test showed significantly lower apoptotic indexes in SM,BEM high,middle,low groups than in A/R group,and in all the SM,BEM high,middle,low groups,the positive expressed Bcl-2 gray scale value increased significantly(P

Antiapoptotic Mechanism of Insulin in Reoxygenation-induced Injury in Cultured Cardiomyocytes of Neonatal Rats / 华中科技大学学报（医学）（英德文版）

To examine the protective effect of insulin on reoxygenation-induced injury and explore the underlying mechanisms, the model of anoxia/reoxygenation (A/R) injury was established by inducing anoxia for 2 h and reoxygenation for 4 h in cultured cardiomyocytes of neonatal rats. The rats were randomized to four groups receiving vehicle, insulin, LY294002, insulin plus LY294002at the onset of reoxygenation after 2 h of anoxia. At the end of reoxygenation of 4 h, activity of lactate dehydrogenase (LDH) and content of malondialdehyde (MDA) were spectrophotometrically determined, apoptosis of cardiomyocytes were detected by using TUNEL and DNA Ladder, and Western blotting was employed to examine the expression of phosphorylated Akt in all groups. Our results showed that compared with vehicle-treated group, activities of LDH, contents of MDA, apoptosis index (AI) were significantly decreased, and expression of phosphorylated Akt was in creased significantly in insulin-treated group. However, changes in LDH, MDA, AI and phospho rylated Akt resulting from insulin were attenuated or abolished by LY294002 (PI3K inhibitor).These data strongly suggest that early administration of insulin at reoxygenation protects cardiomyocytes from reoxygenation-induced apoptosis through PI3K/Akt signaling pathway.

Anoxia-reoxygenation up-regulates transcription of TGF?_1 gene in human umbilical veins endothelial cell line / 重庆医科大学学报

Objective:To investigate the influence of anoxia-reoxygenation on the transcription of TGF?1 gene in Human umbilical veins endothelial cell line(ECV304). Methods:Anoxia-reoxygenation model of Human umbilical veins endothelial cell line was established. TGF?1 mRNA was half-quantitatively analyzed by a modified RT-PCR. Results:The transcription of TGF?1 mRNA was up-regulated significantly in anoxia-reoxygenation group compared with the control group. Conclusion: anoxia-reoxygenation may up-regulate the transcription of TGF?1 gene in human umbilical veins endothelial cell line.

Protective effect of puerarin on anoxia-reoxygenation induced cerebral cell injury in culture / 中国药理学通报

Aim To study the protective effect of puerarin(PUE) on cultured cerebral cells injured by anoxia-reoxygenation. Methods The anoxia-reoxygenation injury model were developed, anoxia for 60 min and reoxygenation for 30 min. the effect of PUE on cerebral ultrastructure was observed.[Ca 2+ ]_i was estimated with Adherent Cell Analysis and Sorting 570(ACAS 570) Laser Cytometer and measured with fluorescent dye Fura-2-AM, the lipid fluidity of cellular membrane was determined by fluorescence polarization technique. Results PUE improved the ultra-structure of cerebral cells, dose-dependently decreased[Ca 2+ ]_i and increased the lipid fluidity of cellular membrane. PUE markedly reduced the chromaticity value of pseudo-colour graphic model of Ca 2+ . Conclusion Puerarin has protective effect on anoxia-reoxygenation induced cerebral cell injury. The effect may be mediated by decreasing[Ca 2+ ]_i and increasing the lipid fluidity of cellular membrane.

Effects of 1,3-dipropyl-8-cyclopentylxanthine,an adenosine A_1 receptor antagonist,on brain neuronal damage induced by hypoxia and reoxygenation / 解放军医学杂志

Objective To observe the effects of 1,3-dipropyl-8-cyclopentylxanthine(DPCPX),an adenosine A1 receptor antagonist,on brain neurons damage induced by hypoxia and reoxygenation(H/R),and to elucidate the relevant mechanisms.Methods An in vitro cultured rat cerebral cortical neuronal H/R damage model was established;the effects of DPCPX were detected at final concentrations of 0(control),25,50,100nmol/L on the lactate dehydrogenase(LDH) release from normoxic neurons and H/R neurons which were treated with hypoxia for 8,12,24 hours followed by reoxygenation for 24 hours;the changes of malondialdehyde(MDA) content,activities of xanthine oxidase(XO) and Ca2+-ATPase in H/R neurons which were treated with hypoxia for 12 hours and reoxygenation for 24 hours brought by administration of DPCPX at the concentration of 100nmol/L were also determined by use of specific reagents.Results With addition of 100nmol/L DPCPX,the LDH release from H/R neurons which were treated with hypoxia for 12 hours and reoxygenation for 24 hours was significantly increased compared with that in control group(P

Effect of Shenmai injection on cardiomyocyte apoptosis after acute anoxia-reoxygenation / 中国病理生理杂志

AIM: To study the effects of Shenmai injection on cardiomyocytes apoptosis after acute anoxia-reoxygenation (A/R) and the possible mechanism. METHODS: In this experiment, cultured cardiomyocytes isolated from neonatal rat were used. Model of myocardial anoxia-reoxygenation injury was produced by depriving oxygen for 5 min and then restoring oxygen for 15 min. The apoptotic cells was detected by flow cytometry to detect labbled Annexin V-FITC/PI. The intracellular calcium level was observed by laser scanning confocal microscopy markered Fluo-3/AM. RESULTS: In anoxia-reoxygenation group, the percentage of apoptotic cells and fluorescent intensity of intracellular calcium were both prominently higher than those in control group (P

Preconditioning with desflurane, sevoflurane and isoflurane increase the preservation of adenosine triphosphate in anoxia-reoxygenation myocardial cells / 中国临床药理学与治疗学

Aim To study the effects of preconditioning with desflurane, sevoflurane and isoflurane on adenosine triphosphate (ATP) in anoxia-reoxygenation myocardial cells. Methods Rat ventricular myocytes, cultured for 4～5 days, were randomly allocated to five groups: Control group, anoxia-reoxygenation group and groups preconditioned with 1.5 MAC desflurane, sevoflurane or isoflurane following anoxia-reoxygenation. The content of intracellular ATP ,the activities of lactic dehydrogenase(LDH) and creatine kinase(CK), and the cell viability were measured at the end of experiment.Results Preconditioning with 1.5 MAC desfllurane, sevoflurane or isoflurane significantly attenuated the great reduction in ATP and cell viability and the increase of LDH and CK caused by anoxia-reoxygenation. There was a positive correlationship between ATP and cell viability,and a negatiue correlationship between LDH and CK (r was 0.83, -0.87 and -0.82 respectively, P