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Objective To investigate the effect of the SMAC gene on paclitaxel sensitivity and cellular activity in lung adenocarcinoma cells based on the caspase-3/Bcl-2/Bax signaling pathway. Methods A paclitaxel-resistant cell line A549/Taxol was established for lung adenocarcinoma, and the cells were divided into four following groups: pcDNA-NC (transfected with pcDNA-NC blank vector), pcDNA-SMAC (transfected with pcDNA-SMAC vector), siRNA-NC (transfected with siRNA-NC empty virus vector), and siRNA-SMAC groups (transfected with siRNA-SMAC lentiviral vector). The SMAC mRNA expression in cells was detected by qRT-PCR; cell sensitivity was detected by MTT; cell proliferation ability was detected by cloning assay; cell invasion ability was detected by Transwell; apoptosis ability was detected by flow cytometry assay; and caspase-3, Bcl-2 and Bax protein expression in cells were detected by Western blot analysis. Results The SMAC mRNA expression was significantly lower in A549 cells compared with BEAS-2B cells (P < 0.05). The SMAC mRNA expression was significantly higher in the pcDNA-SMAC group than that in the pcDNA-NC group cells (P < 0.05). The SMAC mRNA expression was significantly lower in the cells of the siRNA-SMAC group (P < 0.05) than that in the siRNA-NC group. The SMAC mRNA expression was significantly lower in the cells of the siRNA-SMAC group (P < 0.05) than in the siRNA-NC group. Compared with the pcDNA-NC group, the cell IC50, cell clone number, cell invasion ability, and Bcl-2 protein and Bcl-2/Bax ratio were significantly lower in the pcDNA-SMAC group, the cell resistance index reversal was 2.51-fold, and the apoptosis ability and caspase-3, as well as Bax protein expression, were significantly higher (P < 0.05). Compared with the siRNA-NC group, cell IC50, cell clone number, cell invasion ability, and Bcl-2 protein and Bcl-2/Bax ratio were significantly higher in the siRNA-SMAC group, and apoptosis ability and caspase-3 and Bax protein expression were significantly lower (P < 0.05). Conclusion High expression of SMAC increases paclitaxel sensitivity, inhibits cell growth and invasion, promotes apoptosis in lung adenocarcinoma cells, and has a regulatory effect on the caspase-3/Bcl-2/Bax signaling pathway.
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Objective@#To investigate the effect and mechanism of XTP4 gene in apoptotic hepatoblastoma HepG2 cell line.@*Methods@#HepG2 cells were transiently transfected with small interfering RNA of XTP4 genes, plasmid pcDNA3.1/myc-His(-) A-XTP4, and hepatitis B virus X protein transactivated x gene 4 (HBX protein trans-activate gene4, XTP4) and their respective negative controls. After 48h, the overexpression and interference expression condition of XTP4 in HepG2 cells were detected by Western blot. HepG2 cells apoptosis was detected by flow cytometry. The expression levels of apoptosis-related proteins P53, Bcl-2, Bax and Caspase-3 in HepG2 cells were detected by Western blot, and Bcl-2/Bax ratio was calculated. The chemiluminescence assay was used to detect activity of caspase-3 in HepG2 cells. The measured data were presented as ( ± s), and independent sample t-test was used for comparison between the two groups.@*Results@#HepG2 cells had successfully achieved the overexpression and interference expression of XTP4 protein. Compared with the control group, the overexpression of XTP4 in HepG2 cells had significantly decreased the number of apoptotic cells (P < 0.05), and increased Bcl-2/Bax (P < 0.05) ratio, but decreased the expression of P53 protein (P < 0.05). The protein expression of Caspase-3 and activity of caspase-3 was decreased (P < 0.05). However, interference with XTP4 expression in HepG2 cells had significantly increased the number of apoptotic cells (P < 0.05) and decreased Bcl-2/Bax (P < 0.05) ratio, but increased the expression of P53 protein (P < 0.05). The protein expression of Caspase-3 and activity of caspase-3 was increased (P<0.05).@*Conclusion@#In HepG2 apoptosis XTP4 has inhibitory effect, and its effect on inhibiting HepG2 apoptosis may be achieved by regulating the Bcl-2/Bax ratio, and the P53 protein may be involved.
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To explore the protective effect of nerve function of Buyang Huanwu Decoction on cerebral ischemia/reperfusion rats after the transplantation of neural stem cells (NSCs) . Methods Thread bolt method was used to establish middle cerebral artery occlusion model. Drug groups were given Buyang Huanwu Decoction (14. 8 g kg"1 d " 1) by gavage after the rats being sober. NSCs were transplanted to rat brain after making the model 24 hours later. Zea Longa neurobehavioral behavioral score was used to observe neural function defect, and TTC staining to detect the volume of cerebral infarction, and Nissl staining to detect Nissl body integrated optical density (IOD), and Immunohistochemical staining to detect expression of Bcl-2 and Bax. Results Compared with sham operation group, the nerve function defect appeared, and the volume of cerebral infarction increased significantly, the integral optical density of Nissl body was reduced and the ratio of Bcl-2 to Bax was reduced in model group (P < 0. 05) . Compared with model group, the nerve function defect was reduced, the volume of cerebral infarction was reduced, the integral optical density of Nissl body increased, and the ratio of Bcl-2 to Bax increased in BYHWD group, Transplant group and BYHWD + Transplant group (P < 0. 05). Compared with transplant group, the nerve function defect was reduced, the volume of cerebral infarction was reduced , the integrated optical density of Nissl body increased , and the ratio of Bcl-2 to Bax increased in BYHWD + Transplant group (P < 0. 05). Conclusions Buyang Huanwu Decoction can enhance the neuroprotective effect after NSCs transplantation in cerebral ischemia/reperfusion rats.
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Aim To investigate the peptides and its protection for vascular endothelial cells, derived from the absorbed components of rice α-globulin,which was shown to be effective in anti-atherosclerosis. Methods The amino acid sequence was purified by gel chro-matography and RP-HPLC, and determined by ESI/MS. Then the peptide was chemically synthesized. Hu-man umbilical vein endothelial cell injury model was induced by tumor necrosis factor-α. The cell viability was measured by cell counting kit to screen the appro-priate peptide intervention concentration. The apoptotic rate was detected by flow cytometry. Bcl-2, Bax, p-p38, vascular cell adhesion molecule and the protein expression level of NF-κB signaling pathway were de-tected by Western blot and immunofluorescent stai-ning. Results Apoptosis of HUVECs induced by TNF-α was significantly increased by YGEGSSEEG, which also regulated expression of Bcl-2/Bax proteins and inhibited phosphorylation of p38 protein. Besides, the peptide suppressed the production of VCAM-1, ICAM-1 and activation of NF-κB pathway. While it did not significantly improve the oxidative stress response in HUVECs. Conclusion Peptide YGEGSSEEG pro-tects vascular endothelial cells through suppressing ap-optosis and expression of adhesion molecules.
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OBJECTIVES@#To test whether myocardial apoptosis can be induced by traumatic fracture of lower limbs with hemorrhage, in order to lay a foundation of myocardial injury after traumatic fracture for the follow-up study.@*METHODS@#Twenty SD rats were randomly divided into two groups, i. e. control group and trauma group(=10). A rat model of traumatic hemorrhage was establish, and a traumatic model of the original generation of myocardial cell culture was constructed . The level of interleukin-2(IL-2),IL-6,IL-10 and tumor necrosis factor-α(TNF-α) in rat serum was detected by ELISA at 0, 1, 2, 4, 8, 12, 16, 24 and 48 hour to find the most significant point. The pathological cardiac injury in rats was observed by HE staining under a microscope, and the apoptosis of cultured cardiomyocyte was detected by TUNEL methods. The expressions of apoptosis gene,(Bcl-2) and Bax, in myocardium of rat and cultured cardiomyocyte were detected by Western blot and RT-PCR.@*RESULTS@#At the 4 hour after trauma, IL-6 and IL-10 in the serum of rats reached its highest, IL-2 reached its lowest at the 8th hour after trauma, and TNF-αreached its highest at 1 hour after trauma, then all recovered to their normol level gradually. Myocardial HE staining indicated that cardiomyocyte was swelling, disordered derangement, inflammatory cell infiltrated; a large number of myocardial cell nuclei was dyedbrown in TUNEL test which proved that the apoptosis index increased (<0.05). Western blot and RT-PCR results showed that the expression of pro-apoptotic gene Bax was up-regulated (<0. 05), while expression of anti apoptosis gene Bcl-2 down-regulated (<0.05).@*CONCLUSIONS@#The myocardial apoptosis can be induced by traumatic fracture of lower limbs with hemorrhage in rats, and then lead to myocardial injury.
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Animals , Rats , Apoptosis , Cells, Cultured , Cytokines , Blood , Follow-Up Studies , Fractures, Bone , Hemorrhage , Lower Extremity , Pathology , Myocardium , Pathology , Myocytes, Cardiac , Pathology , Proto-Oncogene Proteins c-bcl-2 , Metabolism , Random Allocation , Rats, Sprague-Dawley , bcl-2-Associated X Protein , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To study the effects of allicin on cardiac function and underlying mechanism in rat model of myocardial infarction (MI).</p><p><b>METHODS</b>Ninety-four Wistar rats were randomly assigned to 6 groups (n=14-16 per group): sham control group [underwent thoracotomy without left anterior descending (LAD) occlusion and only received an injection of the same amount of citrate buffer], MI control group (subjected to LAD occlusion and only received an injection of same amount of citrate buffer), positive control group (subjected to LAD occlusion and received an injection of diltiazem hydrochloride at the dose of 1.5 mg/kg), and MI + allicin groups (subjected to LAD occlusion and received an injection of allicin at the doses of 1.2, 1.8, and 3.6 mg/kg). All of the drugs were administered intraperitoneally daily for 21 days. The infarct area was measured by myocardial staining. Hematoxylin-eosin staining was used to observe the pathological changes. Cardiac function parameters were assessed by echocardiography. The myocardial apoptotic index was estimated by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling staining. The expression of Bax and Bcl-2 were detected by quantificational real-time polymerase chain reaction and Western blot.</p><p><b>RESULTS</b>Treatment with allicin could attenuate the myocardial infarct area (P<0.05) and relieve the changes of the myocardium. The left ventricular anterior wall diastolic and systolic thicknesses were increased in the allicin-treated groups (P<0.05), while there was no signifificant difference in the left ventricular posterior wall diastolic and systolic thickness (P>0.05). The left ventricular internal diameter in systole, ejection fraction, fractional shortening, and stroke volume were dramatically elevated in allicin-treated rats (P<0.05). Allicin dose-dependently reduced creatine kinase and lactate dehydrogenase levels (P<0.05). The myocardial apoptotic index was also markedly lowered, and Bax expression was signifificantly decreased, whereas Bcl-2 expression exhibited an opposite trend in allicin-treated rats (P<0.05).</p><p><b>CONCLUSION</b>Allicin appears to exert a cardioprotective effect that may be linked to blocking Bcl-2/Bax signaling pathway-denpendent apoptosis, further improving cardiac function.</p>
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Objective To observe the curative effect of nourishing kidney,activating blood and expelling toxin method Chinese medicine on chronic kidney disease (CKD) ratsand influence on cellular apoptosis.Methods The UUO animal model was adopted.SD male rats were divided into the sham operation group,UUO model group,Chinese medicine low dose group and high dose group.The renal function was measured by the automatic biochemical analyzer,the renal histopathologic change was observed by HE staining,the renal tissue apoptosis rate was observed by TUNEL as well as the protein expression levels of Bcl-2 and Bax were detected by Western blot.Results Compared with the UUO model group,the levels of serum creatinine(Scr) and urea nitrogen(BUN) in Chinese medicine various doses groups were significantly decreased(P＜0.05).In the UUO pathologic observation,renal tubular epithelial cells apoptosis,necrosis,exfoliation,inflammatory cells infiltration were found in the model group,the renal damage in the Chinese medicine various doses groups were slighter compared with the UUO group(P＜0.05).The TUNEL method detection showed that the cellular apoptosis number in the UUO group was significantly increased compared with the sham operation group,the Chinese medicine various doses groups could significantly decrease the apoptosis number(P＜0.05).Compared with the UUO model group,the level of Bcl-2 in the Chinese medicine treatment groups was markedly up-regulated,the Bax level was down-regulated,and the ratio of Bcl-2/Bax was increased(P＜0.05).Above results all showed the dose dependent manner.Conclusion The nourishing kidney,activating blood and expelling toxin method Chinese medicine can obviously down-regulate serum Scr and BUN levels,inhibits the renal cells apoptosis in UUO rat and regulates the Bcl-2 and Bax levels.
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Aim To explore whether total flavonoids of Verbena officinalis L(TFV)can induce apoptosis of HepG-2 cells through targeting topoisomerase Ⅱ.Methods HepG-2 cells were cultured with TFV at 200,100,50 mg·L-1.Cell apoptosis was evaluated by TUNEL-DAPI double staining.TopⅡ activity was detected by supercoiled pBR322 DNA relaxation assay.The levels of the mRNA of TOP Ⅱα,TOP Ⅱβ were analyzed by real time PCR.Expression of Bcl-2,Bax,TOP Ⅱα,TOP Ⅱβ and caspase-3 was analyzed by Western blot.
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Objective To evaluate the effects and underlying mechanisms of crocin on glutamate-induced apoptosis of retinal ganglion cells (RGCs) by affecting extracellular calcium influx.Methods Primary rat retinal ganglion cells were isolated and stimulated with glutamate at concentrations of 0.1 mmol/L and 1 mmol/L for 24 h or 48 h,respectively,to establish apoptosis model of RGCs.Afterwards,crocin of different doses (0.1,1.0 and 3.0 μmol/L) was used to treat the glutamate-induced RGCs for 12 h;then cell apoptosis was detected by Annexin V-FITC/PI staining.The intracellular calcium concentration was determined by FIuo-3/AM fluorescent labeling.Western blot was used to examine the effect of crocin on Ca2+-mediated apoptotic signal molecules calpain and CaMKII.The mitochondrial membrane potential was detected by JC-1 staining and mitochondrial apoptosis-related signaling molecules Caspase-3,Caspase-9 and Bcl-2/Bax were evaluated by Western blot,respectively.Results In comparison with the untreated controls,the cell apoptosis of RGCs exposed to 0.1 mmol/L of glutamate for 24 h did not significantly change (P＞ 0.05).However,apoptosis rate of the cells reached (43.050 ± 2.616) ％ when the stimulation time lasted for 48 h and showed a significant increase (P＜0.01).Treatment with higher-dose glutamate (1 mmol/L) significantly increased apoptosis of RGCs at 24 h (46.450±1.061)％ and 48 h (45.500±3.253)％ compared with the controls (P＜0.01).RGCs were induced by 1 mmol/L of glutamate for 12 h,followed by the treatment with crocin at concentrations of 0.1,1.0 and 3.0 μmol/L,respectively.Each dose of crocin could significantly inhibit cell apoptosis in the dose-dependent manner (P＜0.01).In addition,crocin at 1.0 μmol/L blocked glutamate-induced extracellular calcium influx,inhibited the expression of calcium-dependent proteins Calpainl and CaMK Ⅱ.Moreover,crocin at the dose of 1.0 μmol/L also increased mitochondrial membrane potential,suppressed the expressions of Caspase-3 and Caspase-9,and elevated Bcl-2/Bax ratio.Cornclusion Crocin inhibits glutamate-induced apoptosis of retinal ganglion cells through suppressing extracellular calcium influx,thereby blocking calcium-dependent and mitochondria-dependent apoptosis signaling pathways.
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Aim To study the inhibitory effects of gambogenic acid in combination with miR-218 on cervical cancer HeLa cells and its mechanisms.Methods Eukaryotic expression vector of miR-218(pmi8-218) was transfected into HeLa cells.Transcript levels of miR-218 were quantified by real-time quantitative PCR.HeLa cells were incubated with different concentrations of gambogenic acid alone or in combination with pmiR-218.The cell growth inhibiting ratio of HeLa cells was assessed by MTT assay.Cell apoptosis was measured by fluorescence activated cell sorting.The expression levels of Bcl-2, Bax and E-cadherin were measured by Western blot and qRT-PCR.Results Levels of miR-218 transcript significantly increased in pmiR-218 transfected HeLa cells.Overexpression of miR-218 may enhance the sensitivity of HeLa cells to gambogenic acid.Over expression of miR-218 could enhance the effect of gambogenic acid on inhibition cell proliferation, promoting apoptosis of HeLa cells.pmiR-218 could enhance the regulation of Bax expression and decrease the expression of Bcl-2 in HeLa cells.Conclusions Over expression of miR-218 may enhance the sensitivity of HeLa cells to gambogenic acid;miR-218 can enhance the effect of gambogenic acid on inhibition cell proliferation and promote the apoptosis of HeLa cells, and the mechanism may be related to down-regulation of Bcl-2/Bax expression.
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Objective To observe the curative effect of nourishing kidney,activating blood and expelling toxin method Chinese medicine on chronic kidney disease (CKD) ratsand influence on cellular apoptosis.Methods The UUO animal model was adopted.SD male rats were divided into the sham operation group,UUO model group,Chinese medicine low dose group and high dose group.The renal function was measured by the automatic biochemical analyzer,the renal histopathologic change was observed by HE staining,the renal tissue apoptosis rate was observed by TUNEL as well as the protein expression levels of Bcl-2 and Bax were detected by Western blot.Results Compared with the UUO model group,the levels of serum creatinine(Scr) and urea nitrogen(BUN) in Chinese medicine various doses groups were significantly decreased(P＜0.05).In the UUO pathologic observation,renal tubular epithelial cells apoptosis,necrosis,exfoliation,inflammatory cells infiltration were found in the model group,the renal damage in the Chinese medicine various doses groups were slighter compared with the UUO group(P＜0.05).The TUNEL method detection showed that the cellular apoptosis number in the UUO group was significantly increased compared with the sham operation group,the Chinese medicine various doses groups could significantly decrease the apoptosis number(P＜0.05).Compared with the UUO model group,the level of Bcl-2 in the Chinese medicine treatment groups was markedly up-regulated,the Bax level was down-regulated,and the ratio of Bcl-2/Bax was increased(P＜0.05).Above results all showed the dose dependent manner.Conclusion The nourishing kidney,activating blood and expelling toxin method Chinese medicine can obviously down-regulate serum Scr and BUN levels,inhibits the renal cells apoptosis in UUO rat and regulates the Bcl-2 and Bax levels.
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Objective: To explore whether total saponins of Panax notoginseng (TSPN) can protect hippocampal CA1 subfield neurons against apoptosis following global cerebral ischemia via up-regulating the Bcl-2/Bax ratio and preventing Caspase-3 activation. Methods: Using four-vessel occlusion method to build the global cerebral ischemia model and the ischemia time was 30 min. All rats were divided into Sham group, vehicle group, and different doses (25, 50, 75, and 100 mg/kg) of TSPN groups. The rats in TSPN groups were ip administered with TSPN. The dose of TSPN was suspended in 0.9% saline (10 g/L), while rats in vehicle group were treated with equal volume of 0.9% saline, one injection per day. Compared the survival rate and hippocampal CA1 subfield neuronal density by Nissl staining after reperfusion of 14 d to make sure the best dose of TSPN for neuroprotection. Then to evaluate the neurological score and investigate the expression level of the Caspase-3, Bcl-2, and Bax in the hippocampus CA1 region at days 1, 3, 7, and 14 post-ischemia by immunohistochemistry; In addition, the Western-blotting was adopted to test the protein level of these three proteins. Results: The survival rate of the rats in 75 mg/kg TSPN groups was 100%, and its neuronal density was significantly higher than that in vehicle group and other doses of TSPN groups (P < 0.05); The neurological score in TSPN group was significantly less than that in vehicle group (P < 0.01); The Caspase-3 neuronal density in the CA1 subfield of TSPN group on days 7 and 14 was significantly less than that in vehicle group (P < 0.001); The statistical meaning existed about the protein level of Caspase-3 with molecular weight of 20 000 on days 3, 7, and 14 and 17 000 on days 7 and 14 in two groups (P < 0.001). The neuronal density of Bcl-2 cells in the CA1 subfield and Bcl-2 protein level in the hippocampus of TSPN group at days 7 and 14 was significantly higher than that in vehicle group (P < 0.001); Besides, the Bax neuronal density at days 7 and 14 was significantly lower than that in vehicle group (P < 0.001); And its protein level of the hippocampus was less than that in vehicle group at days 3, 7 and 14 (P < 0.001). The results of ratio of Bcl-2/Bax from not only the neuronal density but also the protein expression demonstrated that the Bcl-2/Bax ratio in TSPN group was significantly higher than that in vehicle groups on days 7 and 14 (P < 0.001). Conclusion: TSPN can protect the hippocampal CA1 subfield neurons against apoptosis following global cerebral ischemia in adult rats via up-regulating the Bcl-2/Bax ratio and preventing Caspase-3 activation.
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OBJECTIVE: To study the apoptosis of human gastric cancer cell line BGC-823 induced by liposome of total glucosides from paeonia(TGP liposome). METHODS: The inhibition of cell proliferation was determined by Cell Counting Kit-8 assay. The morphological changes of the apoptosis cells were observed by HE staining, Hoechst 33258 fluorescent staining, transmission electron microscope. Changes of apoptosis related factor Bcl-2 and Bax protein expression were detected by Western blot. RESULTS: TGP liposome significantly inhibited proliferation of BGC-823 cells in a dose-dependent manner(P < 0.05). GC-823 cells induced by TGP liposome for 48 h was observed on morphological changes such as karyopyknosis and karyorrhexis and apoptotic bodies. Bcl-2 expression was reduced, but Bax expression was increased, so the ratio of Bcl-2/Bax decreased. CONCLUSION: TGP liposome can induce the apoptosis of BGC-823 cells and inhibit its proliferation, its mechanism may be related to down-regulating protein expression of Bcl-2, up-regulating protein expression of Bax, reducing Bcl-2/Bax value.
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Objective To study the reactive oxygen level and the expression of anti-apoptotic protein Bcl-2 and pro-apoptotic protein Bax after treatment of DNA methyltransferase inhibitor 5-aza-2'-deoxycytidine (5-Aza-2'-dc) and paraquat in V79 cells.Methods Cultured V79 cells were divided into 5-Aza-2'-dc treatment group (group A),paraquat treatment group (group B),5-Aza-2'-dc and paraquat treatment group (group C,V79 cells were pretreated with 5-Aza-2'-dc for 12h followed by exposure to paraquat for 12h) and control group (group D).Reactive oxygen level in V79 cells was measured by DCFH-DA flow cytometry and expression of Bcl-2 and Bax was detected by Western blot.Results Reactive oxygen levels and expression levels of Bcl-2 and Bax in V79 cells were significantly different (P＜0.05) in 5-Aza-2'-dc and paraquat treatment group (group C),compared with 5-Aza-2'-dc treatment group (group A),paraquat treatment group (group B) and control group (group D).Expression levels of Bcl-2 and the ratio of Bcl-2 and Bax were lower while reactive oxygen levels and expression levels of Bax were higher in group C than in groups A,B and D.Conclusion 5-Aza-2'-dc regulates DNA methylation by the imbalancing the reactive oxygen metabolism and apoptosis,thus up-regulating the toxic effect of paraquat on V79 cells.
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Aim To investigate the protective effects of sphingosine 1-phosphate ( S1 P ) postconditioning on hypoxia/reoxygenation( H/R) injury in human umbili-cal vein endothelial cells ( HUVEC ) and its mecha-nisms. Methods HUVECs cells were divided into five groups: normal ( control) group, S1P low concentra-tion group ( L ) , S1 P medium concentration group (M), S1P high concentration group ( H) and H/R group. MTT method was used to measure cell survival. Using flow cytometric analysis, the rate of cell apopto-sis was determined. The activities of total superoxide dismutase ( T-SOD) , copper/zinc superoxide dismuta-se ( CuZn-SOD ) , manganese superoxide dismutase ( Mn-SOD) activity, nitric oxide ( NO) and malondial-dehyde ( MDA ) content in cell culture medium were measured with colorimetry. Mitochondrial membrane potential in cells was observed with fluorescence micro-scope. Bax/Bcl-2, eNOS protein expression levels in HUVECs cells were observed with Western blot. Re-sults Compared with H/R group, S1P low, medium and high concentrations in the intervention group could significantly increase the cell survival rate after H/R injury, and increase activity of T-SOD, CuZn-SOD, Mn-SOD and decrease content of MDA. Moreover, S1 P could significantly increase NO content and in-crease eNOS protein expression, decrease apoptosis rate and inhibit the reduction of mitochondrial mem-brane potential. Conclusions S1P can decrease cell apoptosis rate of HUVECs after H/R injury with a cer-tain concentration dependence. The protection of S1P for cell apoptosis of HUVECs after H/R injury may be related to decreasing the intracellular MDA content and improving intracellular SOD activity, increasing mito-chondrial membrane potential and enhancing expres-sion of Bcl-2, anti-apoptotic protein.
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Objective To explore the effect of carnosine in the expression of B cell lymphomal/leukemia-2 (bcl-2) and bcl-2-associated X protein (bax) after focal cerebral ischemia in rats. Methods Thirty male SD rats (SPF scale) were ran?domly divided into 3 groups:sham-operated group, model group and carnosine treated group (n=10 for each group). The mid?dle cerebral artery occlusion model (MCAO) was induced in model group and carnosine treated group. Rats were received carnosine [1 000 mg/(kg·d), orally] in carnosine treated group, and the other rats were received the same volume of normal sa?line (NS) in shame-operated group and model group. The neurological deficit score was used to evaluate the neurological function at 24 h and 72 h after MCAO. Morphological changes were observed by HE staining. TCC staining was used to label infarct volume, and immunohistochemistry was used to detect the expression of bcl-2 and bax. Results Compared with model group, the score of neurological function and infarct volume were significantly declined in carnosine treated group at 72 h after injury (P<0.05 or P<0.01). The changes of ischemic impairment were lighter in carnosine treated group than that of model group. Compared with sham-operated group, the expression levels of bcl-2 and the ratio of bcl-2/bax were de?creased while the expression of bax was increased in model group (P<0.05). Compared with model group, carnosine could sig?nificantly increase the expression of bcl-2 and the ratio of bcl-2/bax, and reduce the expression of bax (P<0.01 or P<0.05). Conclusion Carnosine can enhance bcl-2 expression, decrease bax expression and increase the ratio of bcl-2/bax, which is likely to be one of the mechanisms of neuroprotection.
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Flavonoids are products of secondary metabolism of plants. They are present in herbs and trees and also act as natural chemopreventives and anticancer agents. Ligaria cuneifolia (Ruiz & Pav.) Tiegh., Loranthaceae, is a hemiparasite species that belongs to Argentine flora. Phytochemical studies have disclosed the presence of quercetin, catechin-4β-ol and pro-anthocyanidine as polyphenolic compounds in the active extracts. We previously demonstrated that ethyl acetate extract was capable of reducing cell proliferation and inducing apoptotic death of lymphoid tumor cells. The aim of the current study is to determine whether or not catechin, isolated from L. cuneifolia extracts can induce leukemia cell death and to determine its effect on the cytoplasmatic proteins that modulate cell survival. Our results show that catechin can reduce proliferation of murine lymphoma cell line LB02. The effect is mediated by apoptosis at concentrations upper to 100 µg/mL. Cell death is related to the loss of mitochondrial membrane potential (ΔΨm) and a down regulation of survivin and Bcl-2 together with the increase of pro-apoptotic protein Bax. In summary, the current study indicates that catechin present in the extract of L. cuneifolia is in part, responsible for the anti-proliferative activity of whole extracts by induction of ΔΨm disruption and modulation of the anti-apoptotic proteins over expressed in tumor cells. These results give new findings into the potential anticancer and chemopreventive activities of L. cuneifolia.
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Objective To confirm whether in post-resuscitation myocardial dysfunction involved in myocyte apoptosis mechanism in porcine model of cardiac arrest and apoptosis index varied from different modalities of cardiopulmonary resuscitation or not.Methods A total of 22 WZSP inbred small swine were randomly (random number) divided into sham operation group (SHAM) (n =6),defibrillation first group (DF,n =8) and chest compression first group (CF,n =8).Eight minutes after ventricular fibrillation was set up,standard CPR was carried out subsequently after defibrillation in porcine models of cardiac arrest in DF group and defibrillation after standard CPR in CF group,and hemodynamics were monitored.Twentyfour hours after restoration of spontaneous circulation (ROSC),animals were sacrificed,and myocardial specimens were examined with electron microscopy,Western blot,quantitative RT-PCR,and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay.The experimental data were analyzed by SPSS 17.0.Student's t test was employed for comparisons between two groups.Differences within groups at different time intervals were compared with repeated measures ANOVA.A two-tailed value of P ＜ 0.05 was considered statistically significant.Results Myocardial function was significantly impaired after ROSC.Levels of Bcl-2,Bax and caspase-3 protein was markedly increased in the CF and DF groups than those in the SHAM group (P ＜ 0.05) at 24 h after ROSC,while Bcl-2/Bax was significantly reduced in the CF and DF group compared with the SHAM group (P ＜ 0.05),and much more apoptotic cells were observed in cardiac arrest animals in comparison with sham-operation animals (P ＜ 0.05).Six hours after ROSC,hemodynamic indicators improved significantly in group DF than those in group CF,but Bcl-2,Bax and caspase-3 protein levels and apoptotic index were not significantly different bewteen the DF group and CF group (P ＞ 0.05).Conclusions Caspase-3-mediated apoptosis might be one of the main pathological mechanisms of postresuscitation myocardial injury in a porcine model of cardiac arrest,but there was no statistically significant difference in apoptosis index between two resuscitation modalities,showing no one modality was superior over another.
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@#Objective To observe the effects of Tongxinluo on cerebral ischemic tolerance and expression of Bcl-2 and Bax in rats with cerebral infarction.Methods The neurophysical assessment and the volume of the cerebral infarction in Tongxinluo group,middle cerebral artery occlusion(MCAO)group,preconditioning and MCAO(PC-MCAO)group were determined at different time point.The expression of Bcl-2 and Bax were measured with immunohistochemitry and the apoptosis of the nerve cells were detected with TUNEL.Results Among these groups in the corresponding time,the neurophysical marks of Tongxinluo group was the lowest and the cerebral infarction volume was the smallest at 24 h,3 d and 5 d points.The Bcl-2 positive neurons were the highest,while the Bax positive neurons and the apoptosis rate of nerve cells were the lowest around the necrotic areas in Tongxinluo group(P<0.05).Conclusion Tongxinluo can improve the cerebral ischemic tolerance and increase the expression of Bcl-2 and decrease that of Bax around the necrotic area after cerebral ischemia in rats.
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@#Objective To investigate the possible protective effect of neurotropin on the corresponding spinal cord motoneuron after sciatic nerve injury early in rats.Methods 108 male Sprague Dawley rats were divided randomly into Groups A,B and C with 36 animals in each group,and all animals were cut both side of sciatic nerve,instantly make the operation of end-to-end anastomose of epineurium for abscised sciatic nerve.Group A was control group.Group B was treated with neurotropin and Group C was treated with normal saline after operation.The rats were sacrificed on 1st d,4th d,9th d,14th d,21st d and 28th d after operation.The corresponding spinal cord segments(L4～L6)were gotten to make Bcl-2 and Bax immunohistochemical staining and TUNEL staining.Results The values of Bcl-2/Bax on 9th d,14th d and 21st d after operation of Group B were different from that of Group A and Group C.The number of positive cells stained by TUNEL staining on 9th d was significantly different among Group A,Group B and Group C(v<0.05).The expression of positive TUNEL staining cells reached a peak on 14th d after operation,the number of Group B was less than that of Group A and Group C.Conclusion After instantly make the operation of end-to-end anastomose of epineurium for abscised sciatic nerve,neurotropin can protect the spinal cord motoneurons to avoid apoptosis excessively in some extent.