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Objective To know the Tanshinone Ⅱ A induce human small cell lung cancer cells (H446 cells) apoptosis,and discuss the possible molecular mechanism.Methods Determined by MTT method of different concentrations of Tanshinone Ⅱ A impact on human small cell lung cancer H446 cells proliferation;Determined by Hoechst 33258 method of different concentrations Tanshinone Ⅱ A influence on human small cell lung cancer H446 cells apoptosis;Determined by qPCR method of different concentrations of Tanshinone Ⅱ A impact on human small cell lung cancer H446 cells apoptosis gene;Determined by Westen blot method of different concentrations of Tanshinone lⅡ A effect on lung cancer H446 cells apoptosis.Results Different drug concentrations of Tanshinone Ⅱ A (0.3,0.6,1.25,2.5,5 and 10μg/ml) inhibit human small cell lung cancer H446 cells proliferation and depend by the concentration and time;With the increase of the concentration of the drug in the formation of human small cell lung cancer H446 cells apoptosis corpuscle number increase,significant difference was found in high concentration group (P ＜ 0.05);High concentration group of Tanshinone Ⅱ A promote apoptosis gene (Bax,caspase-9,caspase-3) expression,and inhibit apoptosis inhibiting genes (Akt,the Bcl-2) expression and there were significant differences (P ＜ 0.05);High concentration group of Tanshinone Ⅱ A promoting apoptosis protein (Bax,caspase-9,caspase-3)expression,and inhibit apoptosis inhibiting protein (Akt,the Bcl-2) expression and there were significant differences (P ＜ 0.05).Conclusion Tanshinone Ⅱ A may be through Akt-Bax/Bcl-2-caspase-9-caspase-3 signaling pathways induced human small cell lung cancer H446 cells apoptosis.
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Objective To investigate the etiologic roles of apoptosis-associated genes,environmental factors and their interactions in lumbar disc herniation (LDH).Methods A case-control trial was conducted.We recruited 128 outpatients with LDH as case group and 132 normal people matched by age and gender as control group.Peripheral venous blood samples were collected and DNA was extracted from leukocytes.By using a modified Brucker Autoflex MALDI-TOF mass spectrometer,we analyzed 3 genes with 9 polymorphic sites,namely,Fas-1377G/A rs2234767,Fas-670G/A rs1800682,Fas rs2147420,Fas rs2296603,Fas rs7901 656,Fas rs1 57101 9,FasL-844C/T rs7631 10,CASP-9-1263A > G rs4645978,and CASP-9-712C > T rs4645981.The correlations between polymorphism of Fas,FasL and CASP-9 genes and the risk of LDH were evaluated by non-conditional Logistic regression model.Multiple Logistic regression model was performed to assess the interaction between apoptosis-associated genes and environment factors,such as lumbar vertebral loads,bed type,spare-time exercises and spare-time activities. Results There were preferable balances in case and control groups in age and gender without significant differences.However,the two groups differed significantly (P G (rs4645978),and FasL-844C/T TT and CASP-9-1263A>G GG genotypes might be the high risk genotypes of LDH.The gene-environment interaction analysis revealed that super-multiplicative and sub-multiplicative interactions respectively between FasL-844TT genotype and lumbar vertebral loads (3-4 level),and between CASP-9-rs4645978 GG and lumbar vertebral loads (3-4 level).Conclusion FasL,CASP-9 genes and lumbar vertebral loads and their interactions play important roles in the pathogenesis of LDH.It suggests that the risk of LDH may be codetermined by environmental factors and inherited susceptibility genes,and that the mechanisms of interactions vary in different genotypes and the same or different environmental factors.
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Objective To explore and analyze the effects of allicin prodrug on proliferation of esophageal cancer cell line Eca9706 and the expression of apoptosis gene.Methods Different concentrations of allicin prodrugs were divided into a total of four groups:A1 group(10μg/mL),A2 group(20 μg/mL),A3 group (40 μg/mL),A4 group (normal saline,0 μg/mL),and respectively applicated in esophageal carcinoma cell line Eca9706.After culturing for 1 days,2 days,3 days,cell proliferation was detected by MTT and expression of p53 at the mRNA level was detected by RT-PCR.Results The optimum concentration of proliferation inhibition of allicin prodrug on esophageal cancer cell line Eca9706 was 20μg/mL and the best inhibition time was 2 days;the gene level of esophageal cancer cell line Eca9706 was inhibited with a dose-dependent.Conclusion Allicin prodrugs could effectively inhibit the proliferation of esophageal cancer cell line Eca9706,and control the proliferation of esophageal cancer cells by regulating the expression of apoptosis associated genes,so as to eliminate tumor cells.
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Objective To establish mouse poisoning model by inhaling benzene, and to investigate the induction effect of benzene on the apoptosis of mouse bone marrow cells and its mechanism, and to provide an experimental basis for study on bone marrow toxicity mechanism.Methods 24 male mice were randomly divided into four groups (n=6).The mice in one group were exposed to ambient air (control group)and the mice in the other three groups were exposed to different doses (400,800,1 600 mg·m-3 )of benzene (low,middle and high doses of benzene groups)for 1 5 d in the respective inhalation chambers. At the end of the experiment, the mice were killed. The bone marrow of the mice was obtained. The pathological changes of the bone marrow cells of the mice in various groups were observed under light microscope with HE staining.The apoptotic rates and mitochondrial membrane potential (MMP ) of the mice in various groups were detected by flow cytometry, and the expressions of mitochondrial-deperdent apoptosis related gene proteins were determined with immunohistochemistry method. Results The number of distal and central cells in different doses of benzene groups were significantly reduced,and accompanied by blood sinus expansion in high dose of benzene group.The apoptotic rates of the cells in middle and high doses of benzene groups were obviously higher than that in control group (Ρ<0.01),and there were also significant differences between high dose group and low,middle doses of benzene groups (Ρ<0.05).The MMP was significantly decreased with the increasing of benzene doses, and there were significant differences between middle,high doses of benzene groups and control group (Ρ<0.05).The number of Bax,CytC positive cells in different doses of benzene groups and the number of Caspase-9,Caspase-3 positive cells in middle and high doses of benzene groups were significantly increased compared with control group(Ρ<0.05);the number of Bcl-2 positive cells in different doses of benzene groups was decreased(Ρ<0.05),and number of Bcl-2 positive cells in middle and high doses of benzene groups was decreased compared with low dose of benzene group (P<0.05). Conclusion Benzene with certain dose can induce the apoptosis of mouse bone marrow cells, and promote the expressions of mitochondrial apoptosis related gene proteins. Benzene-induced apoptosis through mitochondrial-dependent apoptosis pathway may be an important mechanism of bone marrow toxicity induced by benzene.
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Objective To investigate the effect of arctigenin on the apoptosis of FBL-3 cells and its mechanisms.Methods The mouse erythroleukemia FBL-3 cells were taken as subjects.The untreated FBL-3 parental cells were taken as the control group,and the FBL-3 cells treated with 20 mg/L arctigenin for 24 h were taken as the experimental group.The effects of arctigenin on the apoptosis of the mouse erythroleukemia FBL-3 cells were determined by agarose gel electrophoresis and flow cytometry assay.The changes of apoptosis-related genes were analyzed by reverse transcriptase-polymerase chain reaction(RT-PCR).Results The agarose gel electrophoresis results revealed that the " DNA ladder" was displayed in the experimental group compared to the control group.The flow cytometry data demonstrated that the apoptosis number of FBL-3 cells in the experimental group was markedly increased compared to that in the control group (t =60.681,P =0.000).The RT-PCR assay results suggested that the expressions of Bcl-2 and IAP-1 gene were down-regulated in the experimental group compared to the control group (t =14.732,29.702,all P =0.000),while the expressions of Bax and Smac gene were up-regulated(t =6.721,8.499;P =0.003,0.001),but the expression of Bcl-XL did not change(t =0.209,P =0.844).Conclusions Arctigenin can induce the apoptosis of the mouse erythrolenkemia FBL-3 cells.Apoptosis of the mouse erythrolenkemia FBL-3 cells induced by arctigenin may be correlated to the down-regulation of Bcl-2 and IAP-1 and up-regulation of Bax as well as Smac.
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Objective To observe the effect of neural stem cell (NSC)transplantation on the hindlimb motor function and caspase-3 expression of motor cortex (MC) in spinal cord transected (SCT) rats. Methods S prague-Dawley (SD) rats were randomly divided into sham operated group, operation group( T9 transection), and subacute NSC transplantation group. The MC of each group( n= 8 )was harvested 7 days post operation (dpo), then western blot was employed to detect the level of caspase-3( β-actin was used as control ). The left 5 animals of each group were subjected to BBB score evaluation at 16th week,then the animals were sacrificed and the MC was harvested and performed immunostain by using caspase-3 rabbit antibody. Results Following NSC transplanta tion, BBB scores( ( 7.58 ± 0. 99 ) ) increased significantly than seen in the SCT animals( ( 5.16 ± 1.19) ). The expressional level of caspase-3 at 7day post operation was( (0.89 + 0.12)) in MC of SCT rats,while it decreased significantly to( (0.76 + 0.11 ) ) in NSC transplantation rats(P＜0.05). The immunoreactive stain of caspase-3 was seen in the cytoplasm of pyramidal neuron in the cortex. Conclusion NSC engraft can downregulate the expression of caspase-3, corresponding to a significant improvement in hindlimb motor function. These findings indicate that NSC transplantation probably regulate the expression of apoptosis genes in MC to promote neurological function recovery in rats subjected SCT.
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Objective To study the effect of PUMA gene mediated by recombinant adenovirus vector combined with radiation on the pancreatic carcinoma. Methods The PANC-1 cells were infected with Ad-PUMA (MOI = 10, 50 and 100, respectively) for 48 h. The expression of PUMA mRNA and protein was detected by RT-PCR and Western blot, respectively. PANC-1 cells were divided into 4 groups: control group, transfection group, irradiation group and combined treatment group. The cell growth inhibition rate and apoptotic rate of PANC-1 cells were assessed by MTT assay and flow cytometry. Human pancreatic carcinomas were transplanted subcutaneously in nude mice, which were randomized into 4 groups: control group, transfection group, irradiation group and combined treatment group. Tumor growth rate and apoptotie index at different time points were recorded in 35 days. Results The expression of PUMA mRNA and protein was increased with the increase of MOI of Ad-PUMA, which was does-dependant (MOI = 10, mRNA = 0.46 ± 0.02, protein = 0. 75 ±0.09;MOI=50, mRNA= 1.12±0.09, protein = 1.01 ±0.18;MOI= 100, mRNA= 1.50±0.08, protein=1.80 ± 0.15 ;P < 0.05). The proliferation of PANC-1 cells was suppressed significantly when transfected by Ad-PUMA in a dose-dependent manner(r = -0.986 55), which was more significant combined with radiation (r = -0.971 26, P < 0.05). Meanwhile, the apoptotie rate was increased in the same manner [for pre- and post-irradiation,which was (45.4 ± 5.26) % and (73.2 ± 6.62) %, respectively, P < 0.05]. From 7 to 35 d after PUMA gene transfection and radiotherapy, the tumor growth was significantly slower than those of irradiation group, transfection group and control group [35 d after therapy, the volume of tumor was (19.82 ± 6.45)mm3 ,(39.5 ± 9.23)mm3 , (33.6 ±3 10.3)mm3 and (52.0 ± 11.43)mm3 , respectively, P < 0.05]. And the apoptotic index was increased in the same manner (AI = 0.43 ± 0.05, 0.29 ± 0.10, 0.24 ± 0.05 and 0.00 ± 0.00, respectively, P < 0.05). Conclusions Recombinant adenoviral-mediated PUMA gene combined with irradiation could increase the cell-killing effect on pancreatic carcinoma. It is better than that of either one kind of therapy.
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@#ObjectiveTo explore the mechanism of estrogen inhibiting osteoblast apoptosis induced with serum hungry.MethodsOsteoblasts of the second or third generation from newly born SD rats calvaria were divided randomly into three groups: control group,serum hungry group,serum hungry with estrogen group.Cells of each group were incubated for 1,2,3,5,7 or 14 d,and then were stained immunohistochemically.The rates of positive cells of each group were analyzed.ResultsThere was a little positive expression of Bax,Bcl2 and Fas in control group.The expression of Bax and Fas were significantly increased(P<0.05)in serum hungry group,peak time was 14 d,but the expression of Bcl-2 were not affected.Compared with that of serum hungry group,the expression of Bax and Fas significantly decreased(P<0.05) in serum hungry and estrogen group,peak time was still 14 d,while that of Bcl-2 increased(P<0.05).ConclusionSerum hungry can increase the expression of Bax and Fas in osteoblast,that can be inhibited by estrogen.Estrogen can also increase the expression of Bcl-2 in osteoblast.All of these may play a role in inhibiting osteoblast apoptosis induced with serum hungry.
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@#ObjectiveTo determine the cerebral protection effect and mechanism of propofol on global cerebral ischemia and reperfusion damage in rats.Methods19 adult male Wistar rats were randomly divided into 3 groups, ischemia group (n=7), propofol group (n=7), and sham injury group (n=5). Global cerebral ischemia and reperfusion model were made by means of Pulsinelli's method. Rats in propofol group were anesthesia with propofol at the dosage of 1.5 ml/h for 30 min at the beginning of reperfusion. Apoptosis and necrosis rate were detected by cytometry. In the same time, bcl-2, Bax and p53 protein expression in hippocampus neurons were detected. ResultsThe apoptosis and necrosis rate in propofol group were significantly decreased as compared with ischemia group ( P<0.05). Bax and p53 protein expression in hippocampus neurons were also significantly decreased as compared with ischemia group (P<0.05), however, no significant findings in bcl-2 protein expression (P>0.05).ConclusionPropofol can decrease apoptosis and necrosis rate in cerebral ischemia reperfusion injured neuron, and the mechanism maybe related to decreasing the expression of Bax, p53 protein.
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Objective: To investigate antisense integrin?V and ?3 gene therapy in Rats mod- el of Pancreatic Carcinoma. Methods: The antisense integrin?V and ?3 expression plasmids wereelectrobloted into pancreatic tumours of rats induced by dimethylbenzanthracine,The tumor inhibit rate was calculated, the tumor microvessel density(MVD)with the immunohistochemical staining and tumor apoptosis with TUNEL were examined. Results: The tumor weight of the control group, ?V group, ?3 group an- d?V?3 group was (1.167?0.79)g(,0.953?0.26)g(,1.013?0.42)g(,0.788?0.56)g respectively. The dif- ference was significant among them(P
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@#ObjectiveTo research the apoptosis of breast cancer cells by application of small direct-current electric field(EF) and its related possible mechanism.MethodsSmall direct-current EFs were applied in human breast cancer cell line MCF7 in vitro to observe the effects on apoptosis of cancer cells by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL)and to detect the mRNA expression of p53, Rb and E2F1 genes by reverse transcriptase-polymerase chain reaction (RT-PCR).ResultsA lot of cancer cells were observed slaughed from culture dish by 200mV/mm of the field strength and the number of the apoptotic cancer cells was gradually significantly increased with the application of EFs(P<0.01). mRNA expression level for p53 and Rb genes was elevated but that for E2F1 gene was decreased overtly in cancer cells exposed to EFs.ConclusionApoptosis of breast cancer cells can be induced by small direct-current EFs and the change for expression of p53, Rb and E2F1 genes may be involved in the mechanism.
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Objective To introduce the relationship between the apoptosis hepatocyte and its genic mediation and the ischemia of portal vein. Methods The combination of related literatures and our research findings were made.Results Portal vein ischemia may induced hepatocyte apoptosis, p53 and bcl-2 gene alternatively adjust hepatocyte apoptosis. Expression of p53 gene is enhanced in hepatic tissue when hepatocyte apoptosis is not obvious, but after 24-72 h of portal vein ischemia, when hepatocyte apoptosis is obvious, enhanced expression of p53 gene or reduced expression of bcl-2 gene occur. There exists close relationship between portal vein ischemia and hepatocyte apoptosis. Conclusion Apoptosis hepatocyte is involved in organic atrophy after ischemia of portal vein, and p53 and bcl-2 gene alternatively adjust hepatocyte apoptosis. At present, the mechanism of apoptosis of hepatocyte induced by ischemia of portal vein is not clear, which needs further study.
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Objective To study the clinical significance of expression of survivin gene in transitional mucosa(TM) adjacent to rectal carcinoma.Methods Mucinhistochemical methods were used to observe the distribution of TM adjacent to rectal carcinoma, and using immunohistochemical methods to observe the expression of survivin gene product in TM,normal mucosa,atypical dysplasia and cancer tissue.Results The positive expression level of survivin gene product was lowest in normal rectal mucosa,and gradually increase in TM,dysplasia and carcinoma tissue(all P
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Objective:To investigate the regulatory mechanism of LBP on T lymphocytes excessive apoptosis and the mRNA expression of apoptotic genes in aged rats Methods:T lymphocyte apoptosis and the expression of pre apoptotic and anti apoptotic genes (Fas, FasL, TNFR1, Bax, Bcl2 and TNFR2) from young rats and old rats treated with or without LBP were studied using flow cytometry and fluorescence real time quantitative RT PCR Results:LBP could effectively downregulate the percentage of T cell apoptosis in aged rats At the same time LBP also downregulated the transcription level of TNFR1 gene and upregulated the transcription level of Bcl 2 in T cells from old rats Conclusion:The T cell excessive apoptosis in old rats could be effectively downregulated with LBP, by the by of decreasing the expression of pro apoptotic gene and enhancing the expression of anti apoptotic gene
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AIM : To investigate the effects of angiotensin II (AngII) in different concentrations and different action times on the rate of apoptosis and expression of apoptosis genes in vascular smooth muscle cells and the influences of fosinopril on it. METHODS : Flow cytometer was used to measure the apoptosis rate and the expression of apoptosis genes (Fas and Bcl 2) induced by AngII in different concentrations and different action times and the influence caused by fosinopril. RESULTS BZ : The rate of apoptosis and the expression of apoptosis genes were induced and increased by AngII with the increase of concentrations and action times compared with that in the control group. The expression of Fas was increased and Bcl 2 was decreased with the increase of the concentrations and action times of AngII in fosinopril group compared with that in the AngII group. CONCLUSION : The apoptosis rate and the expression of apoptosis genes are induced and increased by angiotensin II with the increase of the concentrations and the action times. Fosinopril has the regulation effect on the apoptosis rate and the expression of apoptosis genes in vascular smooth muscle cells.
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Objective To study the effect of mild hypothermia on neuronal apoptosis and Bcl 2,Bax during cerebral ischemia in middle cerebral artery occlusions(MCAO) in rats.Methods The model of MCAO was set up in rats using the Longa's way,and mild hypothermia was treated for 0.5h,1h,3h. After reperfusion(24h) the rat brains were cut into three sections for TUNEL staining and Bcl 2?Bax protein immunohistochemical staining.Results 1)The numbers of apoptotic cells and Bax cells in the mild hypothermia group(1h, 3h) decreased remarkable as compared with the control group( P
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Objective To investigate the possible mechanism of Gongqing Granule(Granule for Clearing the Uterus) on shortening the duration of vaginal bleeding after medical abortion.Methods The 120 unwanted pregnancy women within 49d of intrauterine gestation were randomly divided into three groups.Gongqing Granule combined with mifepristone and misoprostol was administered orally in Group A(treatment group).Qianzhi Capsule combined with mifepristone and misoprostol was administered orally to Group B(control group),and only mifepristone plus misoprostol was administered orally to Group C(blank group).Other 40 women(Group D,induced abortion group),were treated with vacuum aspiration for abortion.Chorionic villi and decidual tissue were collected and expression of Bax and Bcl-2 were detected by immunohistochemical method and Bax mRNA and Bcl-2 mRNA by situ hybridization.Results The expression of Bax,Bcl-2,Bax mRNA and Bcl-2 mRNA was found in all groups.The expression of Bax and Bax mRNA in the treatment group was higher than those in the control group,then the blank group,and then induced abortion group.While the expression of Bcl-2 and Bcl-2 mRNA of the treatment group was lower than those in the control group,then the blank group,and then induced abortion group(P
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Objective To study the effect and the mechanism of sibelium on apoptosis and transcription of bcl 2 clan and interleukin 1? converting enzyme(ICE) gene in limbic system after rats' middle cerebral artery occlusion(MCAO).Methods MCAO model was made, and determined the dynamic changes on the cell counts of apoptosis, bcl 2 clan and ICE gene positive transcription in temple ortex, hippocampus and hypothalamus in the controls of normal saline,sibelium 50 ?g intervenient group and sibelium 100 ?g intervenient group during rats MCAO or reperfusion.Results The apoptosis counts of every times in sibelium 100?g intervenient group was significant lower than another two groups( P 0.05).The bcl 2 gene positive transcription cell counts at every times in sibelium 100?g intervenient group were significant higher than another two gruops( P