Resveratrol Attenuates Hydrogen Peroxide-induced Injury of Rat Ovarian Granulosa-lutein Cells by Resisting Oxidative Stress via the SIRT1/Nrf2/ARE Signaling Pathway.

INTRODUCTION: This paper aims to reveal the molecular mechanism of resveratrol against oxidative stress and cell injury. The ovarian granulosa-lutein cell injury and apoptosis induced by oxidative stress may be responsible for female luteal phase deficiency. The antioxidant function of resveratrol has been confirmed; however, its effect on the expression of antioxidant enzymes and regulatory mechanisms in ovarian granulosa-lutein cells remains unclear. OBJECTIVE: This study aimed to investigate the role of the SIRT1/Nrf2/ARE signaling pathway in the effect of resveratrol on the hydrogen peroxide-induced injury of rat ovarian granulosa-lutein cells. METHODS: In this study, ovarian granulosa-lutein cells extracted from 3-week female SD rats were treated with 200 µM H2O2 in the presence or absence of 20 µM resveratrol. siRNA-SIRT1 and siRNA-Nrf2 were used to inhibit the expression of SIRT1 and Nrf2, respectively. Cell counting kit 8 (CCK-8), cellular morphology, progesterone secretion, and estradiol were used to evaluate cell injury. Hoechst 33258 staining was used to measure cell apoptosis. DHE staining, DCFH-DA staining, malondialdehyde content, protein carbonyl content, total antioxidant capacity and SOD viability were used to estimate the levels of oxidative stress. Western blot analysis was used to detect the levels of apoptosis-related proteins, and SIRT1/Nrf2/ARE signaling pathway-related proteins. RESULTS: The H2O2 treatment-induced rat ovarian granulosa-lutein cells injury was shown as decreased cell viability, impaired cellular morphology, and decreased levels of progesterone and estradiol. The H2O2 treatment also exacerbated cell apoptosis demonstrated as more apoptotic cells stained by Hoechst staining, decreased level of anti-apoptosis protein Bcl-2 and increased level of pro-apoptosis protein Bax. These effects of cell injury and apoptosis induced by H2O2 can be ameliorated by resveratrol. Resveratrol also alleviated oxidative stress induced by H2O2, supported by decreased superoxide anion and cellular total ROS, decreased malondialdehyde and protein carbonyl levels, and increased total antioxidant capacity and SOD viability. Western blot results demonstrated resveratrol reversed the H2O2-induced decrease in levels of antioxidant enzymes containing ARE sequences and activated SIRT1/Nrf2 pathway. Further treatment by siRNA-Nrf2 suggested resveratrol could not activate the expression of antioxidant enzymes under a condition of inhibition of Nrf2. CONCLUSION: This study demonstrates that resveratrol attenuated oxidative stress to protect H2O2-induced rat ovarian granulosa-lutein cell injury and apoptosis via SIRT1/Nrf2/ARE signaling pathway.

Synthetic Analogs of Marine Alkaloid Aplysinopsin Suppress Anti-Apoptotic Protein BCL2 in Prostate Cancer.

Aplysinopsins are a class of indole alkaloids that possess various pharmacological activities. Although their action has been studied in regard to many diseases, their effect on prostate cancer has not yet been examined. Therefore, we synthesized a new series of aplysinopsin analogs and investigated their cytotoxic activity against prostate cancer. Five analogs showed high antitumor activity via suppressing the expression of the anti-apoptotic gene Bcl2, simulationously increasing the expression of the pro-apoptotic genes p53, Bax and Caspase 3. The inhibition of BCL2 led to the activation of BAX, which in turn activated Caspase 3, leading to apoptosis. This dual mechanism of action via apoptosis and cell cycle arrest induction is responsible for aplysinopsin analogs antitumor activity. Hence, our newly synthesized analogs are highly promising candidates for further preclinical studies against prostate cancer.

Design and synthesis of quinoxaline-1,3,4-oxadiazole hybrid derivatives as potent inhibitors of the anti-apoptotic Bcl-2 protein.

Quinoxaline is one of the privileged heterocyclic fragments for drug molecules. Quinoxaline anticancer drug candidates XK469 and CQS exhibit antiproliferative and proapoptotic properties against various cancers. Based on their chemical structures, we therefore synthesized a series of quinoxaline-1,3,4-oxadiazole hybrids and assessed their anticancer potential on human leukemia HL-60 cells. Although these hybrids exerted significant inhibition of HL-60 cell proliferation, they showed high cytotoxicity on human normal cells (WI-38). Utilizing information from molecular modelling of the hybrids to the anti-apoptotic Bcl-2 protein, we added substructures including phenyl, piperazine, piperidine, and morpholine rings to their frameworks. The designed quinoxaline-1,3,4-oxadiazole hybrid derivatives successfully induced apoptotic response on HL-60 cells with low toxicity on WI-38 cells. Furthermore, RT-PCR analysis demonstrated that these derivatives predominantly inhibit Bcl-2 expression. Our findings highlight the great potential for the development of synthetic quinoxaline-1,3,4-oxadiazole hybrid derivatives as proapoptotic anticancer agents.

Knockdown of miR-429 Attenuates Aß-Induced Neuronal Damage by Targeting SOX2 and BCL2 in Mouse Cortical Neurons.

Accumulation of amyloid-ß peptide (Aß) and massive neuronal death due to apoptosis were the essential steps in the pathogenesis of Alzheimer's disease (AD). MiR-429 was reported to play an important role in the pathogenesis of AD. However, the detailed function and underlying molecular mechanism of miR-429 in the pathogenesis of AD remain elusive. Cortical neurons were stimulated with 20 µM of Aß25-35 for 24 h to construct AD model in vitro. qRT-PCR assay was used to detect the expression of miR-429, and qRT-PCR or western blot analysis were performed to assess the levels of Sex-determining region Y-box 2 (SOX2) and B cell lymphoma-2 protein (BCL2) at mRNA or proteins levels in the AD mouse model and Aß-induced treated cortical neurons. Luciferase reporter assay and western blot analysis were used to confirm the potential targets of miR-429. CCK-8 assay, flow cytometry analysis, and caspase3 activity assay were used to measure cell viability, cell apoptosis capacity and caspase3 activity, respectively. MiR-429 was upregulated and SOX2 and BCL2 were downregulated in the AD mouse model and Aß-induced mouse cortical neurons. MiR-429 knockdown attenuated Aß-induced cytotoxicity in mouse cortical neurons. SOX2 and BCL2 were direct targets of miR-429. Moreover, anti-miR-429-mediated neuroprotective effect was abated by the restoration of SOX2 or BCL2 expression. Knockdown of miR-429 might attenuate Aß-induced cytotoxicity by targeting SOX2 and BCL2 in mouse cortical neurons, providing a novel prospect in AD therapy.

Apoptotic and anti-apoptotic seromarkers for assessment of disease severity of non-alcoholic steatohepatitis.

BACKGROUND AND STUDY AIMS: This study aimed to find out non-invasive markers for the assessment of severity of non-alcoholic steatohepatitis (NASH) in an attempt to decrease the need for liver biopsy. It also aimed to evaluate the key role of apoptosis in the pathogenesis of the disease and the suggested role of anti-apoptotic factors in therapeutic modalities and disease prognosis. PATIENTS AND METHODS: The serum levels of soluble Fas (s. Fas), s. Fas ligand, cytokeratin 18 (CK-18) fragment and Bcl-2 were measured in 80 patients and 15 non-hepatic subjects as control. The patients were divided based on histological examination of liver biopsy into three groups. Group I included 40 patients with NASH, group II had 40 patients with non-alcoholic fatty liver disease (NAFLD) non-NASH and group III had 15 non-hepatic subjects as control. Apoptosis of hepatocytes was assessed by morphological examination using a light microscope and expressed as number per square millimetre. RESULTS: There was a significant increase in the serum levels of s. Fas, s. Fas ligand and CK-18 fragments in the NASH group. The anti-apoptotic protein Bcl-2 showed significantly low levels in NASH patients. Apoptosis of hepatocytes was significantly higher in the NASH group. The degree of apoptosis was inversely correlated with the level of Bcl-2. A significant correlation between both s. Fas and CK-18 fragment with liver histology with regard to lobular inflammation and ballooning was found. CONCLUSIONS: Increased serum levels of s. Fas and CK-18 fragment in the NASH group and its correlation with the severity of disease suggested the key role of apoptosis in NASH pathogenesis which can be used for the assessment of the severity of NASH. A high level of anti-apoptotic Bcl-2 in NAFLD suggests its protective role in disease progress.

Effect of ischemic preconditioning on myocardial Bcl-2 expression and mitochondrial structure during heart valve replacement surgery / 中国病理生理杂志

AIM: To investigate the effect of ischemic preconditioning (IP) on myocardial Bcl-2 expression and mitochondrial structure during heart valve replacement surgery under cardiopulmonary bypass. METHODS: Fifty-four patients were prospectively randomized to receive or not ischemic preconditioning (IP) before cold cardioplegic arrest. Ischemic preconditioning in the IP patients (n=22) was induced by a single 2-min ischemia followed by 3-min reperfusion just before aortic clamping and cold crystalloid cardioplegia for myocardial protection. The control group (n=32) received no ischemic preconditioning before cold cardioplegic arrest. The levels of ejection fraction (EF), fractional shortening(FS) and stroke volume (SV) in both groups were measured and compared. troponin T (c-TnT) level, Bcl-2 protein expression and microscopic changes of myocardial mitochondrial structure were recorded for each group before and after surgery. RESULTS: The level of EF, FS and SV in IP group was higher than those in control group (P<0.05). No significant difference in preoperative c-TnT levels between two groups was observed. The level of c-TnT in IP group was lower than that in control group and with a declining trend over time of 6 h, 24 h, 48 h, 72 h and 5 d after surgery, respectively. The preoperative positive unit of Bcl-2 expression between two groups showed no statistical difference (P> 0.05). Postoperatively, the positive unit of Bcl-2 expression in IP group was 19.85±5.88, significantly increased as compared to the preoperative value (P<0.05). In control group, the positive unit of Bcl-2 expression was 14.17±3.39, showed no statistically significant difference to the preoperative value (P>0.05). Postoperative Bcl-2 expression between two groups showed a significant difference (P<0.05). In the control group, microscopic observation revealed swollen mitochondrion, with a hardly visible or disrupted membrane for some mitochondrion;mitochondrial crista were obviously dissolved and loose with a large number of vacuoles formation. However in IP group, myocardial mitochondrion appeared with intact membrane, concentrated mitochondrial cristae with high electron density and no vacuoles formation was observed. CONCLUSION: IP may up-regulate the expression of myocardial anti-apoptotic protein Bcl-2 to protect the mitochondrion, thus protecting cardiocytes and cardiac functions.

Cinobufacini induces the apoptosis of U937 cells and its mechanism / 肿瘤

Objective: To investigate the effect of cinobufacini on apoptosis of U937 cells and its possible mechanism. Methods: Cell viability was measured by MTT assay. The morphological changes were observed by Wrightś staining and immunofluorescence staining. DNA fragmentation was visualized by agarose gel electrophoresis. Apoptotic rate was evaluated by teminal deoxynucleotidyl transferase (TdT) labeling in situ. Expression of bel-2 protein was analyzed by flow cytometry. The levels of Fas and Fas-1 mRNA were measured by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR). Results: Compared with the control group, treatment with cinobufacini at 0.225 to 1.8 μg/ mL for 1-3 d significantly inhibited growth of U937 cells in a time and dose dependent manner. The IC50 value was 1.36 μg/ mL at 24 h. The typical morphological changes of apoptosis and typical apoptotic DNA ladder were observed after incubation with cinobufacini at 0.9 μg/mL for 1 d. The apoptotic rate evaluated by TdT labeling in situ were 4.8%, 13.57%, and 24.33% after exposure to cinobufacini at 0.225, 0.45, and 0.9 μg/ mL for 1 d, respectively. The expression levels of bcl-2 protein and Fas-1 mRNA significantly decreased and the expression of Fas mRNA significantly increased in apoptotic cells. Conclusions: Cinobufacini inhibits growth and inducs apoptosis of U937 cells via inhibition of bcl-2 and Fas-1 genes and activation of Fas gene.

Enhancing synaptic plasticity and cellular resilience to develop novel, improved treatments for mood disorders.

There is mounting evidence that recurrent mood disorders - once considered "good prognosis diseases"- are, in fact, often very severe and life-threatening illnesses. Furthermore, although mood disorders have traditionally been conceptualized as neurochemical disorders, there is now evidence from a variety of sources demonstrating regional reductions in central nervous system (CNS) volume, as well as reductions in the numbers and/or sizes ofglia and neurons in discrete brain areas. Although the precise cellular mechanisms underlying these morphometric changes remain to be fully elucidated, the data suggest that mood disorders are associated with impairments of synaptic plasticity and cellular resilience. In this context, it is noteworthy that there is increasing preclinical evidence that antidepressants regulate the function of the glutamatergic system. Moreover, although clearly preliminary, the available clinical data suggest that attenuation of N-methyl-D-aspartate (NMDA) function has antidepressant effects. Recent preclinical and clinical studies have shown that signaling pathways involved in regulating cell survival and cell death are long-term targets for the actions of antidepressant agents. Antidepressants and mood stabilizers indirectly regulate a number of factors involved in cell survival pathways, including cyclic adenosine monophosphate (cAMP) response element binding protein (CREB), brain-derived neurotrophic factor (BDNF), the antiapoptotic protein bcl-2, and mitogen-activated protein (MAP) kinases, and may thus bring about some of their delayed long-term beneficial effects via underappreciated neurotrophic effects. There is much promise for the future development of treatments that more directly target molecules in critical CNS signaling pathways regulating synaptic plasticity and cellular resilience. These will represent improved long-term treatments for mood disorders.

Apoptosis and Bcl-2 protein expression in rat kidney tissue after severe burn with delayed fluid resuscitation in areas of different altitude / 解放军医学杂志

Objective To observe the changes in Bcl-2 protein expression in rat kidney tissue after severe burn with delayed fluid resuscitation in areas of different altitude,and explore the relationship between its expression and cell apoptosis.Methods The experiments were performed at two altitudes(1 517m and 3 848m).A 30% TBSA Ⅲ? scald model was reproduced with 240 male Wistar rats(120 rats for each altitude).Rats were randomly assigned into delayed fluid resuscitation group(DFR,n=50),immediate fluid resuscitation group(IFR,n=60) and normal control group(NC,n=10).Renal tissue samples were harvested at 1,6,12,24,72 and 168 hours after scald,respectively.The cell apoptosis was detected by tissue chips technic and terminal uridin nick end labeling(TUNEL).The expression of Bcl-2 was detected by immunohistochemistry and image analysis.Results In higher altitude,cellular edema,granular degeneration,necrosis and disintegration of renal tubular epithelial cells;dilation and engorgement of renal glomerular capillaries,degeneration and necrosis of endothelial cells,and congestion,edema and inflammatory cell infiltration in renal interstitium were found.The pathological changes were more serious in DFR group than that in IFR group,and they were worse in the rats at 3 848m altitude than that in those rats at 1 517m altitude.The levels of cell apoptosis and Bcl-2 protein expression were higher in DFR group than that in IFR group,and in the rats at 3 848m altitude than that in those rats at 1 517m altitude(P

Relationship between CD44v6 and Bcl-2 protein expression in typroid cancer / 中国免疫学杂志

Objective:In order to study the correlation between The expression of CD44v6 protein and Bcl-2 protein in thyroid cancer were examined.Methods:The expression of CD44v6 and Bcl-2 in 50 thyroid cancers,20 adjacent noncancerous portion,45 adenoma and 10 normal thyroid tissue were respecitively investigated by catalyzed signal amplification immunohistochemical technique. Results:The positive rate of CD44v6 and Bcl-2 in thyroid cancer was 64.0% and 46.0%, which was significantly higher than that in adenoma or adjacent noncancerous ( P

Apoptosis and Bcl-2/Bax expression in the early follicles of reproductive women / 中国病理生理杂志

AIM: To investigate the apoptosis and Bcl-2/Bax expression in the early follicles of women at reproductive age. METHODS: 12 ovarian specimens were collected from reproductive women (aged 23-38 years) undergoing gynaecological operation. Histopathological examination of these specimens was performed to confirm its' morphological normalities. Using TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling) assay and immunohistochemistry method, cell apoptosis and Bcl-2/Bax expression were examined in the early follicles including mainly primordial, intermediary and primary follicles. RESULTS: 18.75% of the oocytes were found TUNEL positive in the early follicles, but no granulosa cells in these follicles were found TUNEL positive. Bax expression was detected in 76.07% of the oocytes in the early follicles, but Bcl-2 expression was negative in these oocytes. In addition, Bcl-2/Bax expression were not present in the granulosa cells in early follicles. CONCLUSION: The oocyte apoptosis occurs in the early follicles of reproductive woman, and pro-apoptotic protein Bax may play a role in regulating this process. It suggests that Bax mediated oocyte apoptosis may be the molecular mechanism of the early follicle atresia in the ovaries of reproductive woman. [

Inhibitory effect and induction of apoptosis of caffeic acid Ge on growth of U14 in mice / 中国病理生理杂志

AIM:To evaluate the antitumor effect of caffeic acid Ge on U14 tumor bearing mice.METHODS:The tumor inhibitory ratios of caffeic acid Ge on the growth of U14 in mice was observed.Apoptosis morphological transformation of U14 cells induced by caffeic acid Ge was detected by electronic scan microscope and MG-P staining.Alteration of cell cycle was analyzed by flow cytometry.Apoptosis-related protein levels of Bax and Bcl-2 were determined by immunity histochemistry technology.MTT assay was applied to study the antitumor activities of caffeic acid Ge in U14 cell lines in vitro.RESULTS:Tumor inhibitory rates in caffeic-acid Ge groups were 38.50%,47.17% and 64.02%(from low dose to high dose)(P

Relationship between the Expression of Apoptosis-Related Proteins and Chemosensitivity in Gastric Cancer Cell Lines

BACKGROUND: There has been a growing realization that a variety of anticancer drugs can induce apoptotic cell death. In the present study, an attempt was made to investigate the responsiveness of gastric cancer cells to various anticancer drugs and to identify which apoptosis-related proteins could be correlated to chemosensitivity. METHODS: Nine human Korean gastric cancer cell lines (SNU-1, -5, -16, -484, -601, -620, -638, -668, and -719) were analyzed. The cytotoxicity of each cell line to camptothecin, cisplatin, mitomycin C, vincristine, 5-FU, epirubicin, and doxorubicin was determined by using a MTT (dimethylthiazole- diphenyltetrazolium-bromide) assay. Apoptosis-related proteins (p53, p21, Bcl-2, Bcl-x, and Bax) were detected using a Western blot assay. RESULTS: Of the nine gastric cancer cell lines, SNU-1 was resistant while SNU-5 was sensitive to anticancer drugs. Mutated p53 was detected in all the cell lines. The highest expression of Bcl-2 was observed in SNU-1 while less or no expression of Bcl-2 was observed in SNU-5, -484, and -601. Bcl-xL was less expressed in SNU-5 than in the other cell lines. CONCLUSIONS: Chemosensitivity in gastric cancer cell lines was correlated mainly with the level of Bcl-2 and partly with that of Bcl-xL. There was no correlation between the chemosensitivity and other apoptosis-related proteins, such as p21, p53, Bax, and Bcl-xS in the studied gastric cancer cell lines.

Role of bcl-2 gene expression in inhabition of hepatocellular carcinoma by genistein in nude mice / 中国病理生理杂志

AIM: To probe into the role of 1,4,5-trisphosphate inositol(IP3) and bcl-2 gene expression in inhibiting hepatocellular carcinoma of nude mice by genistein.METHODS: Animals with hepatocellular carcinoma were treated with genistein 1 mg?kg-1?d-1(ip) for 3 weeks.The volume and weight of tumaor were measured.IP3,bcl-2 mRNA,Bcl-2 protein were assayed by IP3- Birtrak assay,RT-PCR,Western blotting,respectively.RESULTS: The tumor volume and weight of animals treated with genistein were lower than those in control(42.7mm3?27.8mm3 vs 52.3mm3?26.5mm3,42.7mg?27.8 mg vs 91.3mg?31.4 mg).IP3 content was lower than that in control [(13.4?1.4)nmol/g protein vs(35.3?6.6)nmol/g protein].bcl-2 mRNA expression was lower in group treated with genistein than that in control(RI which was the gray degree multiply area of bcl-2 / the gray degree multiply area of ?-actin 0.48?0.02 vs 0.56?0.15).Bcl-2 protein expression was lower in group treated with genistein than that in control(RI 1.69?0.52 vs 1.37?0.48).CONCLUSION: Genistein inhibits growth of transplanted hepatocellular carcinoma in nude mouse liver by reducing IP3 production and down-regulating bcl-2 gene expression.

Bcl-2 AND BAX PROTEIN EXPRESSION IN HEPATOCELLUAR CARCINOMAS / 中国现代普通外科进展

Objective: To investigate the expression of Bcl-2, the apoptosis inhibitor, and Bax, the apoptosis promoter in hepatocellular carcinomas (HCCs). Methods: 35 formalin-fixed and paraffin-embedded samples of histologically confirmed HCC, were examined using immunohis to chemical staining for Bcl-2 and Bax protein. Results : 7 of 35 HCCs were positive for Bcl-2 protein and 13 of 35 positive for Bax protein. Immunoreactivity for Bcl-2 protein was present in 24 of 35 HCC cases and Bax protein in 25 of 35 cases. Furthermore, all positive cells were cytoplasmic stained. 6 of 7 Bcl-2 protein positive HCCs were (histological grade Ⅱ ), Bax protein expression had little association with histological grading. Co-expression of Bcl-2 and Bax protein was only present in 4 cases. At any rate, a significant high Bcl-2 and Bax protein expression was detected in small HCCs. Also, Bcl-2 protein expression tended to decrease with clinical stage. Conclusion: Bcl-2 and Bax immunohistochemical expression in HCCs seems to be associated with histological grading. Also, Bcl-2 protein seems to be relative to tumors with good prognosis. It supports the hypothesis that Bcl-2 expression has prognostic value.

Effect of bone marrow stromal cells on the anoxia cardiomyocytes in vitro / 中国病理生理杂志

AIM: To investigate the influence of bone marrow stromal cells (BMSCs) on the anoxia cardiomyocyte apoptosis. METHODS: Using the anaerobic culture apparatus, the apoptosis of the cardiomyocytes, the BMSCs alone and co-cultured with each other were detected by morphological observation, PI staining flowcytometry, electrophoretic gel mobility analysis of DNA fragmentation. Western blotting was used to detect Bax and Bcl-2 protein expression. RESULTS: Compared with control, the BMSCs were unsensitive to anoxic cultured while the anoxic cardiomyocytes were prone to apoptosis. Apoptosis of cardiomyocytes was increased significantly, detected by PI staining and agarose gel elestrophoresis showed “DNA ladder”. However, when anoxia cardiomyocytes co-cultured with BMSCs, apoptosis cells were decreased, “DNA ladder” disappeared and the expression of protein Bax was also decreased. CONCLUSION: Bone marrow stromal cells prevent the anoxia cardiomyocytes from apoptosis, probably by suppressing the expression of Bax protein.

Mitochondrial ceramidase overexpression up-regulates Bcl-2 protein level in K562 cells probably through its metabolic product sphingosine-1-phosphate / 中国病理生理杂志

AIM: To investigate the role of mitochondrial ceramidase in mitochondrial functions, especially in the regulation of apoptosis. METHODS: pCDNA3.1/His-MtCDase plasmid, containing mitochondrial ceramidase cDNA sequence, was transfected into K562 cells by liposome, and G418 was used to screen the positive clones. A stable transfected K562 cell line was established and defined as ‘K562TC’. The differences between K562 and K562TC cells in serum withdrawal resistance and Bcl-2 protein expression were evaluated by annexin V/PI test, flow cytometry and Western blotting, respectively. RESULTS: K562TC cells with elevated Bcl-2 protein expression level identified by FCM or Western blotting showed stronger resistance to apoptosis induced by serum withdrawal than their parental cells. Inhibition of mitochondrial ceramidase expression in K562TC cells by its specific antisense oligodeoxynucleotide was correlated with a decrease in Bcl-2 protein level. N, N'-dimethylsphingosine (DMS), a sphingosine kinase inhibitor, depleted intracellular sphingosine-1-phosphate (SPP) production, also abrogated Bcl-2 protein expression in K562TC cells, while exogenous sphingosine-1-phosphate up-regulated Bcl-2 protein level in K562 cells. CONCLUSION: Mitochondrial ceramidase overexpression in K562 cells leads to markedly elevated level of Bcl-2 protein and results in more resistance to serum withdrawal. This effect is initiated not by sphingosine, the direct metabolite of mitochondrial ceramidase, but via sphingosine-1-phosphate, its phosphorylated form, indicating that mitochondrial ceramidase, through its sphingoid metabolite sphingosine-1-phosphate, up-regulates Bcl-2 protein expression in K562 cells.

Protective effect and the possible mechanism of Nano-Se on myocardium of experimental diabetes mice / 中国病理生理杂志

AIM:To observe the protective effect of Nano-Se on myocardium of experimental diabetes mice.METHODS:Sixty healthy male KM mice were chosen,ten of which were selected randomly as the normal control group.After fasted for 24 h,the rest 50 mice were injected with streptozotocin(STZ,50 mg/kg)intraperitoneally for 5 d.At 7th d,the blood-sugar was measured from vena caudalis,40 mice,of which blood-sugar exceeded 16.65mmol/L,were selected and randomized into 4 groups:the positive control group,low dose(25 ?g/kg)Nano-Se group,mid dose(50 ?g/kg)Nano-Se group,high dose(50 ?g/kg)Nano-Se group.All mice were given intragastric administration of 0.2 mL normal saline and corresponding dose of Nano-Se.The body weights were measured every week,and the dose of which was adjusted according to the change of the body weights.8 weeks later,the mice were killed and cardiac muscle of the left ventricle was taken.The myocardium was prepared to 10% homogenate for measuring SOD,GSH-Px activity and MDA content.The myocardial cell apoptosis was measured by TUNEL.The expressions of Bc1-2 and Bax proteins were determined by immunohistochemistry.RESULTS:Compared to normal group,the SOD and GSH-Px activities in positive control group decreased,MDA level increased,the rate of myocardial cell apoptosis increased significantly,Bc1-2 protein expression deceased and Bax protein expression increased.Compared to positive control group,the SOD and GSH-Px activities in low and mid dose Nano-Se groups expression increased,MDA level decreased,myocardial cell apoptosis rate decreased,Bc1-2 protein expression increased and Bax protein expression decreased.Moreover,the SOD and GSH-Px activities in high dose Nano-Se group decreased obviously compared to those in mid dose Nano-Se group.MDA level and myocardial cell apoptosis rate increased,Bc1-2 protein expression decreased and Bax protein expression increased,no significant difference in SOD,GSH-Px activity,MDA level and myocardial cell apoptosis rate was observed compared with positive control group.CONCLUSION:The damage of cardiac muscle is alleviated when a certain dose of Nano-Se is supplied to diabetes mice.The protective mechanism may be related to antioxidation,blood-sugar adjustment and the increase of Bc1-2 expressing.

Senescence of endothelial cells and gene expression associated with apoptosis induced by angiotensinⅡ / 中国病理生理杂志

AIM:To study the senescence of human umbilical vein endothelial cells(HUVECs) and Bcl-2,Bax gene expression associated with apoptosis induced by angiotensinⅡ(AngⅡ).METHODS:HUVECs were cultured in vitro and the cell viability was observed by methyl thiazolyl tetrazolium(MTT).HUVECs were intervened by AngⅡ and valsartan(AngⅡ type 1 receptor blocking) and divided into 3 groups:the control group,AngⅡ group(stimulated with AngⅡ10-6mol/L for 48 h),valsartan group(valsartan was added to cells 1 h before 10-6mol/L AngⅡ treatment).?-gal staining aod cell cycle analysis were used to identify the cell aging status.Morphologic changes and percentage of apoptosis were assayed with Hoechst33258 under fluorescent microscope.The expressions of Bcl-2 and Bax,and the apoptosis-associated genes were detected by immunocytochemical staining,RT-PCR and Western blotting.RESULTS:The cell viability by AngⅡ-induced cells was(81.9%?4.1)%,the positive cell number of ?-gal staining was significantly higher in AngⅡ-induced cells(80.10%?6.81)% than that in the control cells.The cell cycle was at G0-G1(91.36%?6.45)%,the apoptotic cells significantly increased(31.84?2.86)% under fluorescent microscope.In valsartan group,Bcl-2 mRNA and protein expression increased markedly(P

Inhibitory effect of hypoxia preconditioning on hypoxia/ reoxygenation-induced apoptosis in cardiomyocytes / 中国病理生理杂志

AIM: To study the role of hypoxia precondit io ning (HP) in hypoxia-reoxygenation (HR)-induced apoptosis in neonatal rat cardio myocytes and the possible mechanisms. METHODS: Cultured neonatal rat cardiomyocytes were divided into three groups: normal group, HP+H/R group and H/R group. Acridine orange (AO) sta ining was performed to detect morphological changes of apoptotic cells. Apoptosi s rates of cardiomyocytes were detected by flow cytometry. Colorimetric assay wa s used to detect caspase-3 activity. Expression of Bcl-2 protein was detected by immunohistochemistry combined with computer image analysis. RESULTS: Apoptotic cells were detected by AO staining after hypo xia of 6 h followed by 3 h-reoxygenation. The hypodiploid apoptotic peak was det ected by flow cytometry with the apoptotic rates of (29.7?5.4)%. A significan tly reduced apoptotic rates of (7.8?1.3)% was detected in HP group(P