Elemene reversed the multidurg resistance of A549 /DDP lung cancer cells via mitochondrial apoptosis pathway

目的:探讨榄香烯乳(ElEMEnE,ElE)逆转人肺腺癌耐顺铂(CISPlATIn,ddP)细胞A549/ddP的耐药性及作用机制。方法:采用MTT法检测榄香烯乳单用的细胞毒作用及与ddP合用时耐药逆转作用。荧光探针JC-1结合激光共聚焦显微镜检测线粒体膜电位的变化。dCfH-dA荧光探针结合流式细胞仪检测细胞内活性氧(rEACTIVE OXygEn SPECIES,rOS)水平。用谷胱甘肽试剂盒结合分光光度法检测计算gSH/(gSSg+gSH)比值。蛋白质印迹法检测胞质中CyTO C、PrO-CASPASE-3、CASPASE-3和bCl-2家族蛋白表达情况。结果:不同浓度榄香烯乳抑制A549/ddP细胞株生长,呈时间-剂量依赖性效应,联合顺铂能提高A549/ddP细胞株对顺铂的敏感性而逆转耐药。不同浓度榄香烯乳联合顺铂使A549/ddP细胞株线粒体膜电位下降,rOS浓度增加,gSH/(gSSg+gSH)比值降低,上调胞质中CyTO C、CASPASE-3、bAd蛋白表达,下调PrO-CASPASE-3、bCl-2蛋白表达。结论:榄香烯乳逆转A549/ddP细胞株耐药性可能与其损伤线粒体膜,活化胞内氧化还原体系,诱导线粒体凋亡路径有关。Objective: To explore the mechanism that elemene( ELE) reversed the multidurg resistance( MDR)of A549 /DDP lung adenocarcinoma cell.Methods: MTT assay was used to determine the growth inhibition of human lung adenocarcinoma A549 /DDP cells in vitro.Mitochondrial membrane potential( MMP) was monitored by JC- 1fluorescence probe with laser confocal scanning microscopy,the intercellular reactive oxygen species( ROS) level was measured by 2',7'- dichlorfluorescein- diacetate( DCFH- DA) staining and flow cytometry and the ratio of GSH /( GSSG +GSH) was calculated according to detection results of GSH kit.The expression of Cytochrome C,Caspase-3 and the Bcl- 2 family proteins and in the case of cyclosporine A and DEVD- CHO,the expression of Caspase- 3expression were measured by Western blot.Results: MTT results showed that different concentrations ELE could inhibit the proliferation of A549 /DDP cells in a time- and dose- dependent manner.Intriguingly,ELE plus cisplatin enhanced the sensitivity of A549 /DDP cells to cisplatin and reversed A549 /DDP cells dury resistance.Different concentrations ELE decreased mitochondrial membrane potential,increased intracellular ROS concentration and decreased GSH /( GSSG + GSH) ratio of A549 /DDP cells in a time- and dose- dependent manner.Furthermore,the combination with both ELE and cisplatin also enhanced the protein expression of Cytoplasmic C,Caspase- 3 and Bad,and reduced the protein levels of Bcl- 2 and Pro- caspase- 3 in the cisplatin- resistant A549 /DDP cancer cells.Conclusion: ELE reversed the MDR of A549 /DDP cell line may demage mitochondrial membrane,active intracellular redox system and induce the mitochondrial apopotosis pathway.福建省自然科学基金面上项目(编号:2010D014

The efect of elemene reversing the multidurg resistance of A549/DDP lung cancer cells

目的 :探讨榄香烯乳(ElEMEnE)逆转人肺腺癌耐顺铂(CISPlATIn,ddP)细胞A549/ddP的耐药性及可能的作用机制。方法 :采用MTT法检测榄香烯乳单药作用时的细胞毒作用及与ddP联合作用对A549/ddP细胞耐药的逆转作用;HOECHST 33342法进行细胞核染色,荧光显微镜下观察细胞形态的改变;fCM检测细胞凋亡率;蛋白质印迹法检测细胞质中细胞色素C(CyTOCHrOME C,CyT C)、PrO-CASPASE-3、CASPASE-3和抗凋亡因子b细胞淋巴瘤-2(b CEll lyMPHOMA-2,bCl-2)家族蛋白表达的情况。结果 :不同浓度榄香烯乳对A549/ddP细胞均有一定的抑制作用,呈时间-剂量依赖性效应,联合ddP能提高耐药细胞株A549/ddP对ddP的敏感性而逆转其耐药性。HOECHST 33342法和fCM法检测结果显示,榄香烯乳联合ddP能更大程度上诱导A549/ddP细胞的凋亡,提高耐药细胞的凋亡率,同时还上调A549/ddP细胞质中CyT C、CASPASE-3和bAd蛋白的表达,而下调PrO-CASPASE-3和bCl-2蛋白的表达。结论 :榄香烯乳能逆转A549/ddP细胞耐药性可能与其损伤线粒体膜,使线粒体释放CyT C到细胞质、同时活化CASPASE-3、上调促凋亡蛋白bAd和下调抗凋亡蛋白bCl-2表达以启动凋亡路径有关。Objective: To investigate the effect of elemene on the drug resistance of cisplatin(DDP)- resistant human lung adenocarcinoma A549/DDP cells, and its possible mechanism.Methods: The growth inhibition of A549/DDP cells treated with elemene alone or combined with different concentrations of DDP was detected by MTT assay.The morphological changes of apoptosis of A549/DDP cells treated with different concentrations of elemene(20 and 40 μg/mL) for 24 h were observed by Hoechst 33342 staing under fluorescence a microscope.The apoptosis rate of A549/DDP cells treated with different concentrations of elemene(20 and 40 μg/mL) after 24 h were detected by FCM(flow cytometry).The expressions of cytochrome C, pro-caspase-3, caspase-3 and the Bcl-2 family proteins were measured by Western blotting analysis.Results: MTT result showed that different concentrations elemene could inhibit the proliferation of A549/DDP cells in a time- and dose- dependent manner.Intriguingly, elemene plus DDP enhanced the sensitivity of A549/ DDP cells to DDP and reversed the resistance of A549/DDP cells.Elemene was also a strong inducer of apoptosis in this model system, and a synergistic effect on induction of cell death was observed when the tumor cells were treated with both agents.The result showed that elemene could enhance A549/DDP cell apoptosis.Furthermore, the combination of elemene and DDP also enhanced the protein expressions of cytochrome C, caspase-3 and Bad, and reduced the protein levels of Bcl-2 and pro-caspase-3 in the A549/DDP cells.Conclusion: The reversal of the multi-drug resistance of A549/DDP cell line by elemene may result from its effect on induction of apoptosis.Elemene treatment can impaire mitochondrial membrane, release cytochrome C into cytoplasm, activate caspase-3,up-regulate pro-apoptotic protein Bad and down-regulate anti-apoptotic protein Bcl-2, and finally caused apoptosis.福建省自然科学基金面上项目(编号:2010D014

The effect of elemene on reversal multidurg resistance and the expression of P-glyco-protein in A549/DDP

目的:探讨榄香烯乳(ElEMEnE,ElE)逆转人肺腺癌A549/ddP细胞株耐药性及其机制。方法:采用MTT法检测榄香烯乳单用的细胞毒作用及与顺铂(CISPlATIn,ddP)合用时耐药逆转作用;采用流式细胞术检测榄香烯乳对A549/ddP细胞内罗丹明-123(rHOdAMInE-123,rH123)蓄积的影响;采用WESTErn blOT检测榄香烯乳对耐药细胞A549/ddP细胞膜上P-gP蛋白表达的影响。结果:不同浓度榄香烯对A549/ddP细胞均有一定的抑制作用,呈时间-剂量依赖性效应。单用ddP的IC50为15.46μg/Ml,合用20μg/Ml榄香烯24H,IC50为4.15μg/Ml,逆转耐药倍数为3.63。同时,榄香烯增加A549/ddP细胞内rH123的蓄集,降低细胞膜上PgP的表达,且呈剂量依赖性,表明榄香烯能减少A5 4 9/ddP细胞对药物的外排和抑制P-gP蛋白的表达。结论:榄香烯在体外逆转肿瘤细胞耐药性可能与其抑制P-gP的功能和表达有关。Objective: To investigate the elemene( ELE) reversal of human lung adenocarcinoma A549/DDP cell resistance and its mechanism.Methods: Cytotoxicity of ELE alone or in combination with DDP in A549 / DDP cells was determined by MTT assay.Based on flow cytometric technology,the effect of ELE on the uptake of rhodamine-1 2 3( Rho1 2 3) in A5 4 9 / DDP cells was detected by measuring Rho123 mean fluorescence intensity( MFI).P- gp protein expression on the A549 / DDP cell membrane were detected by Western blot.Results: Different concentration elemene inhibited the A549 / DDP cell proliferation in a time- dose- dependent manner( P < 0.05).Alonely used DDP,IC 50 was 15.46μg / ml; combined 20μg / ml elemene for 24h,DDP IC 50 was 4.15μg / ml,the reversal activity was 3.63 times higher than that of DDP.Elemene increased intracellular accumulation of Rh123 of the A549 / DDP cells, reduced the expression of P- gp on the cell membrane,in a dose- dependent manner,indicating that elemene reduce A549 / DDP cells for drug efflux and inhibition of P- gp protein pump function.Conclusion: The reversal activity of DDP is possible related to inhibition of P- gp function and expression in vitro,which lead to an increased intracellular accumulation of anticancer drugs.福建省自然科学基金面上项目(编号:2010D014

Calcium channel α_1 expression and significance of axillary sweat glands in primary palmar hyperhidrosis along with axillary hyperhidrosis patients

目的探讨钙离子通道α1在原发性手汗症并腋窝多汗患者腋窝汗腺组织中的表达情况。方法选择2011年10月至2012年9月在我院确诊为原发性手汗症并腋窝多汗的患者28例和同期正常对照组8例,通过手术时切取患者腋窝皮肤汗腺组织1块,显微镜下观察汗腺组织结构,采用rEAl TIME PCr技术和WESTErn blOT蛋白印迹技术测定钙离子通道α1在原发性手汗症并腋窝多汗患者腋窝汗腺组织中的表达情况。结果光镜下两组汗腺组织结构无差异,1.25万倍电镜下可见手汗症组汗腺分泌颗粒数量明显高于对照组,差异有统计学意义(P<0.05)。原发性手汗症并腋窝多汗患者腋窝汗腺组织中钙离子通道α呈高表达,其亚单位α1MrnA和蛋白水平均高于对照组,差异有统计学意义(P<0.05)。结论原发性手汗症并腋窝多汗患者腋窝汗腺组织中汗腺分泌颗粒数量明显增多,钙离子通道α1表达增高,这些可能是手汗症的发病原因之一。Objective To investigate the expression level of the calcium channelα1 in patients with primary palmar hyperhidrosis along with axillary hyperhidrosis.Methods We choosed 28 patients who were diagnosed as primary palmar hyperhidrosis along with axillary hyperhidrosis from October 2011 to September 2012 in our hospital and 8 cases of normal control group during this period.When the patients were operated,we cut out a block of axillary skin sweat gland tissue.We observed sweat gland structure by light and electron microscope,and used Real time PCR technology and Western blot protein imprinting technology to determine the expression level of calcium channelα1 in patients with primary palmar hyperhidrosis along with axillary hyperhidrosis.Results Sweat gland structure had no difference by light microscope.The sweat gland secretion granules of experimental group were obviously higher than that of control group by 12 500 times electron microscopy.The expression levels of calcium channelα1 mRNA and its protein were higher in patients with primary palmar hyperhidrosis along with a axillary hyperhidrosis than those in normal group,and the difference was statistically significant(P<0.05).Conclusion Sweat gland secretion granules obviously increased,the expression level of calcium channelα1 increased in the axillary sweat gland tissues of the patients with primary palmar hyperhidrosis along with axillary hyperhidrosis,these might be one of the causes of primary palmar hyperhidrosis.国家自然科学基金(81070906); 福建省自然科学基金项目(2013J01303); 福建医科大学重大科研项目计划(09ZD016); 福建医科大学苗圃科研基金(2010MP006

Aripiprazole versus other atypical antipsychotics for schizophrenia

BACKGROUND: In most western industrialised countries, second generation (atypical) antipsychotics are recommended as first line drug treatments for people with schizophrenia. In this review we specifically examine how the efficacy and tolerability of one such agent - aripiprazole - differs from that of other comparable second generation antipsychotics. OBJECTIVES: To evaluate the effects of aripiprazole compared with other atypical antipsychotics for people with schizophrenia and schizophrenia-like psychoses. SEARCH METHODS: We searched the Cochrane Schizophrenia Group Trials Register (November 2011), inspected references of all identified studies for further trials, and contacted relevant pharmaceutical companies, drug approval agencies and authors of trials for additional information. SELECTION CRITERIA: We included all randomised clinical trials (RCTs) comparing aripiprazole (oral) with oral and parenteral forms of amisulpride, clozapine, olanzapine, quetiapine, risperidone, sertindole, ziprasidone or zotepine for people with schizophrenia or schizophrenia-like psychoses. DATA COLLECTION AND ANALYSIS: We extracted data independently. For dichotomous data we calculated risk ratios (RR) and their 95% confidence intervals (CI) on an intention-to-treat basis based on a random-effects model. Where possible, we calculated illustrative comparative risks for primary outcomes. For continuous data, we calculated mean differences (MD), again based on a random-effects model. We assessed risk of bias for each included study. MAIN RESULTS: We included 12 trials involving 6389 patients. Aripiprazole was compared to olanzapine, risperidone and ziprasidone. All trials were sponsored by an interested drug manufacturer. The overall number of participants leaving studies early was 30% to 40%, limiting validity (no differences between groups).When compared with olanzapine no differences were apparent for global state (no clinically important change: n = 703, 1 RCT, RR short-term 1.00 95% CI 0.81 to 1.22; n = 317, 1 RCT, RR medium-term 1.08 95% CI 0.95 to 1.22) but mental state tended to favour olanzapine (n = 1360, 3 RCTs, MD total Positive and Negative Syndrome Scale (PANSS) 4.68 95% CI 2.21 to 7.16). There was no significant difference in extrapyramidal symptoms (n = 529, 2 RCTs, RR 0.99 95% CI 0.62 to 1.59) but fewer in the aripiprazole group had increased cholesterol levels (n = 223, 1 RCT, RR 0.32 95% CI 0.19 to 0.54) or weight gain of 7% or more of total body weight (n = 1095, 3 RCTs, RR 0.39 95% CI 0.28 to 0.54).When compared with risperidone, aripiprazole showed no advantage in terms of global state (n = 384, 2 RCTs, RR no important improvement 1.14 95% CI 0.81 to 1.60) or mental state (n = 372, 2 RCTs, MD total PANSS 1.50 95% CI -2.96 to 5.96).One study compared aripiprazole with ziprasidone (n = 247) and both the groups reported similar change in the global state (n = 247, 1 RCT, MD average change in Clinical Global Impression-Severity (CGI-S) score -0.03 95% CI -0.28 to 0.22) and mental state (n = 247, 1 RCT, MD change PANSS -3.00 95% CI -7.29 to 1.29).When compared with any one of several new generation antipsychotic drugs the aripiprazole group showed improvement in global state in energy (n = 523, 1 RCT, RR 0.69 95% CI 0.56 to 0.84), mood (n = 523, 1 RCT, RR 0.77 95% CI 0.65 to 0.92), negative symptoms (n = 523, 1 RCT, RR 0.82 95% CI 0.68 to 0.99), somnolence (n = 523, 1 RCT, RR 0.80 95% CI 0.69 to 0.93) and weight gain (n = 523, 1 RCT, RR 0.84 95% CI 0.76 to 0.94). Significantly more people given aripiprazole reported symptoms of nausea (n = 2881, 3 RCTs, RR 3.13 95% CI 2.12 to 4.61) but weight gain (7% or more of total body weight) was less common in people allocated aripiprazole (n = 330, 1 RCT, RR 0.35 95% CI 0.19 to 0.64). Aripiprazole may have value in aggression but data are limited. This will be the focus of another review. AUTHORS' CONCLUSIONS: Information on all comparisons are of limited quality, are incomplete and problematic to apply clinically. Aripiprazole is an antipsychotic drug with a variant but not absent adverse effect profile. Long-term data are sparse and there is considerable scope for another update of this review as new data emerges from the many Chinese studies as well as from ongoing larger, independent pragmatic trials