槐耳清膏诱导人直肠癌HR8348细胞凋亡的实验研究

目的　探讨槐耳清膏在体外对人直肠癌HR83 48细胞的生长抑制作用和凋亡诱导作用及其作用机理。方法　将处于对数生长期的HR83 48细胞用含槐耳清膏的培养基培养 3 6h ,用四氮唑蓝 (MTT )比色法检测吸光度 (OD)值 ,计算抑制率 ;用甲基绿 派若宁染色法及TUNEL法检测细胞凋亡指数 ;用免疫组织化学方法检测bcl 2、bcl xl、bax、bak及 p5 3基因表达的情况。 结果　槐耳清膏对HR83 48细胞的抑制率随浓度增加而上升 ,浓度为 4.0mg/ml时抑制率最大 (71.1% ) ,与 5 FU组 (浓度为 10 μg/ml)相比差异无显著性意义 (P>0 .0 5 )。甲基绿 派若宁染色法和TUNEL法均可见到典型的细胞凋亡、凋亡小体或出泡现象 ,凋亡指数随槐耳清膏浓度的增加而增加 ,当浓度达 4.0mg/ml时 ,凋亡指数迅速上升 ,甲基绿 派若宁法为 0 .162 0± 0 .0 12 8,TUNEL法为 0 .2 612± 0 .0 15 8,均大于 5 FU组的凋亡指数(0 .0 780± 0 .0 0 95和 0 .10 2 8± 0 .0 13 1) ,P<0 .0 5。槐耳清膏组bcl 2、bcl xl、bak、p5 3蛋白表达较空白对照组明显增强(P<0 .0 5 ) ,而bax变化不明显 (P>0 .0 5 )。槐耳清膏组bak/bcl 2和bak/bcl xl比值显著大于空白对照组。结论　槐耳清膏在体外对人直肠癌HR83 48细胞具有显著的抑制作用和凋亡诱导作用。槐耳清膏     

五羟黄酮调节肝癌HepG2细胞bax基因表达诱导细胞凋亡的实验研究

目的:探讨三磷酸肌醇( IP3) 和bax基因表达变化在五羟黄酮(Quercetin)诱导肝癌细胞凋亡中的作用。方法:以肝癌HepG2 细胞培养72 h为对照,以20，40，60，80μmol/L Quercetin作用于 HepG2 细胞72 h 时和60μmol/L Quercetin 作用于 HepG2 细胞 6，12，24，48，72 h，应用同位素试剂盒检测细胞IP3含量,RT-PCR分析bax mRNA表达，Western blotting 分析细胞bax蛋白表达,流式细胞仪检测细胞凋亡率。结果:各浓度的Quercetin作用于肝癌HepG2 细胞72 h，IP3含量显著低于对照组(P<0.01)，bax mRNA和bax蛋白表达显著高于对照组，细胞凋亡率显著高于对照组(P<0.01); 60 μmol/L Quercetin 作用于肝癌HepG2 细胞6，12，24，48，72 h,各时相IP3含量显著低于对照组(P<0.01）;12 h后bax mRNA和bax蛋白表达显著高于对照组，24 h后各时相细胞凋亡率为显著高于对照组（P<0.01）。结论:Quercetin能减少IP3生成,上调bax基因表达,诱导肝癌细胞凋亡。     

生长抑素类似物诱导胆管癌细胞凋亡和bcl-2/bax表达的研究

目的探讨生长抑素类似物SMS2 0 1 995 (SMS)对人胆管癌SK ChA 1细胞凋亡和凋亡调控基因bcl 2和bax的影响。方法用DNA凝胶电泳、流式细胞仪检测AnnexinV标记凋亡细胞、免疫组化和原位杂交方法研究SMS(10 0ng/ml)处理 6、12和 2 4h后人胆管癌SK ChA 1细胞凋亡和bcl 2 /bax表达的变化。结果SMS作用SK ChA 1细胞 6h对DNA无明显影响 ,作用 12和 2 4h后DNA凝胶电泳出现呈典型梯状条带。SMS作用 6、12和 2 4h后 ,AnnexinV标记的凋亡细胞分别为 (2 2±5 ) % ,(39± 7) %和 (5 8± 10 ) %。同时 ,SMS可使细胞bcl 2蛋白表达下降 ,使bax蛋白和mRNA表达升高。结论SMS能诱导SK ChA 1细胞发生凋亡 ,bax和bcl 2凋亡基因介导了SMS对SK ChA 1细胞凋亡的发生     

bcl-2 mRNA、baX mRNA表达在三氧化二砷诱导肝癌细胞凋亡中的意义

目的 分析三氧化二砷(As2O3)诱导肝癌细胞凋亡中凋亡相关基因bcl—2 mRNA、bax mRNA表达的意义。方法 采用电镜、流式细胞仪、电泳及半定量RT—PCR方法检测不同剂量三氧化二砷诱导人肝癌细胞株bel—7402后凋亡的出现及bcl-2mRNA、bax mRNA的表达。结果 0．5にmol/L As2O3处理肝癌细胞未见凋亡，8μmol/L As2O3诱导肝癌细胞出现明显凋亡。对照组和药物组未检测到bcl-2 mRNA的表达，随浓度增加，bax mRNA的表达上调。结论 As2O3通过诱导凋亡抑制肝癌细胞株bel-7402的增殖，凋亡诱导基因bax mRNA的表达上调与此过程有关。bcl—2mR—NA表达与此过程无关。     

H_2O_2诱导肝细胞凋亡时p53、bcl-2mRNA的表达及其意义

目的　探讨H2 O2 诱导人类正常肝细胞系 (L0 2细胞 )凋亡时p5 3、bcl 2mRNA表达的意义及丹参有效成分Rxa对其表达的影响。方法　以正常L0 2细胞为对照 (L组 ) ,以 10 μmol/LH2 O2 诱导L0 2细胞凋亡为细胞模型 (LH组 ) ,观察Rxa处理 (LaH组 ,Rxa 2mmol/L)对 p5 3、bcl 2mRNA表达 (原位杂交结合图像分析处理 )的影响。结果　LH组 2～ 6hp5 3mRNA表达随凋亡细胞指数增高而增强 ,6～ 8h表达下降 ;bcl 2mRNA表达弱。相比较LaH组各时限检测点p5 3mRNA表达均低于LH组 ,bcl 2mRNA表达较强 (P <0 .0 1)。结论　Rxa可能通过 p5 3的细胞周期调控、DNA损伤修复作用和bcl 2的抗凋亡作用减轻L0 2细胞过氧化损伤。     

阻塞性黄疸大鼠肝组织细胞凋亡及相关基因bcl-2、bax的表达

目的　探讨阻塞性黄疸肝损害的调控机制。方法　用末端脱氧核苷酸转移酶介导生物素标记(TUNEL)技术和免疫组织化学方法检测 4 0只阻塞性黄疸大鼠肝脏组织细胞凋亡状态及凋亡相关基因bcl 2和bax蛋白的表达。结果　胆总管结扎后 ,随结扎时间的延长细胞凋亡增加 ,结扎 14d后细胞凋亡达高峰 ,凋亡指数 (AI)达 5 8.2 3± 1.5 8,各组AI差异有显著性 (P <0 .0 5 )。在阻塞性黄疸过程中 ,bcl 2蛋白表达越强 ,bax蛋白表达越弱 ,AI亦越少。结论　bcl 2和bax蛋白均参与了阻塞性黄疸肝组织中细胞凋亡的调节。     

骨关节炎软骨细胞凋亡调控基因的研究

目的　比较分析正常人及老年性骨关节炎患者软骨细胞bax和bcl 2的表达及细胞凋亡状况。　方法　取 9例骨关节炎患者的关节软骨做实验标本 ,以 6例无骨关节炎病史的意外死亡者关节软骨作为正常对照 ;采用逆转录 /聚合酶链反应 (RT PCR)方法检测bax和bcl 2mRNA表达 ,免疫组化检测bax和bcl 2蛋白 ;应用TUNEL方法进行凋亡细胞原位检测。　结果　骨关节炎患者和正常对照软骨细胞都能表达bax和bcl 2mRNA ;骨关节炎关节软骨细胞baxmRNA表达量较正常对照显著增高 (P <0 0 1) ,bcl 2mRNA表达量也高于正常对照组 (P <0 0 5 ) ,两组间bax/bcl 2表达量的比值差异无显著性意义 (P >0 0 5 ) ;免疫组化可检测到相应表达水平的蛋白 ;骨关节炎软骨细胞凋亡 (4%～ 14% )多于正常对照 (0～ 2 % )。　结论　软骨细胞凋亡受bax和bcl 2共同调节 ;bax和bcl 2的共同调节结果可能是OA患者软骨细胞凋亡增加 ,但凋亡率不高、病理过程进展缓慢的一个重要的原因     

细胞凋亡与增殖在先天性胆总管囊肿发病中的作用

目的　研究不同类型 (囊状、梭状型 )先天性胆总管囊肿壁细胞凋亡与增殖的关系 ,了解细胞凋亡 /增殖在胆总管囊肿中的作用。方法　以胆总管囊状扩张组 32例、梭状扩张组 35例及胆总管结石致胆管扩张组 (对照组 ) 2 5例为研究对象。术中取部分胆总管壁送检 ,观察胆总管壁损伤情况。用免疫组织化学法检测细胞凋亡的相关指标bcl 2、bax及细胞增殖指标PCNA的表达 ;用TUNEL法检测细胞凋亡情况。结果　囊状扩张组胆总管壁粘膜上皮细胞破坏严重 ,梭状扩张组镜下所见与囊状扩张组相似 ,但程度较轻。对照组胆总管粘膜上皮细胞破坏最轻 ,其凋亡细胞呈散在分布 ,上皮细胞和成纤维细胞凋亡阳性率较低 ,分别为 (2 .74± 1 .0 0 ) %和 (2 .95± 0 .87) % ;bcl 2、bax表达率较低 ,水平相似 (P>0 .0 5) ;PCNA表达率较高 ,分别为 (3 .74± 1 .0 0 ) %和 (3 .71± 1 .77) %。囊状扩张组与梭状扩张组囊壁残留上皮中的bcl 2和PCNA表达率低 ,分别为 (0 .99± 0 .51 ) %和 (0 .90± 0 .38) % ;bax呈高表达 ,bcl 2呈低表达 ,凋亡细胞阳性率较高 ,分别为 (1 3 .94± 4 .77) %和 (7.51± 3 .46) % ;成纤维细胞中的PCNA表达率较高 ,分别为 (9.91± 2 .91 ) %和(9.70± 3 .1 8) % ,bax呈低表达 ,bcl 2呈高表达 ,凋亡细胞阳性率较低     

bcl-2和bax基因与细胞凋亡的关系及在脊髓损伤中的作用

目的探讨bcl2和bax基因与细胞凋亡的关系及在脊髓损伤中的作用。方法采用Allen法挫伤大鼠T10节段脊髓,随机分为7组,其中6组为实验组,1组为对照组,每组在SCI后2、4、8、24h、3、7d取材,每一时间点4只。免疫组织化学法检测bax和bcl2基因表达,采用原位末端标记法检测凋亡细胞。结果正常组大鼠脊髓中未见凋亡细胞。实验组在损伤2h出现阳性细胞,伤后8h阳性细胞数目达高峰为84.53±8.00,以后逐渐减少。bax基因表达的高峰也在伤后8h为0.311±0.012,说明bax基因促进了细胞的凋亡,而此时bcl2基因开始出现表达增多,至伤后24h表达达高峰,之后缓慢降低,而与此相对应的是细胞凋亡开始逐渐减少(P<0.01)。结论凋亡相关基因bax与bcl2可能参与了脊髓继发性损伤,细胞凋亡在脊髓继发性损伤过程中起着关键性作用。     

凋亡抑制蛋白bcl-2/bax基因活性水平在乳腺癌发展中的调节作用

我们利用具有表达突变型 p5 3乳腺癌伴有肺转移模型 ,通过α 干扰素诱导凋亡 ,观察细胞内bcl 2和bax的蛋白水平变化 ,探讨 p5 3诱导的细胞凋亡中凋亡抑制蛋白bcl 2 /bax基因所起的作用。一、材料与方法1.材料 :健康雌性津白小鼠 2只 ,8周龄 ,体重 18～ 2 0g ,天津市肿瘤研究所动物室提供。可移植性小鼠乳腺癌(BCML TA2 99)模型 ,由天津肿瘤所实验肿瘤室建立并传代。bcl 2、bax、p5 3免疫组织化学碱性磷酸酶 (SABC)试剂盒(即用型 )及 3 ,3’ 二氨基联苯胺 (DAB)显色剂、磷酸盐缓冲液 (PBS)均购自天津灏祥生物制品科技有限责任公司。α…     

二十二碳六烯酸联合5氟尿嘧啶对胃癌细胞生长及bcl-2、bcl 2l12和bax表达的影响

目的 评估二十二碳六烯酸(DHA)联合5氟尿嘧啶(5-Fu)对胃癌细胞SGC 7901生长以及bcl-2、bcl 2l12和bax基因表达的影响.方法 采用台盼蓝拒染方法检测两药物单用和联用对细胞活力的影响,用联合系数判断两药合用效果,倒置显微镜下观察细胞生长状况,PI染色流式细胞术检测亚二倍体峰比并拟合细胞周期曲线,Annexin-V/PI双标记方法检测早期细胞凋亡,RT-PCR方法检测bcl-2、bcl 2l12和bax基因的表达.结果 DHA可抑制胃癌SGC 7901细胞生长,且呈剂量和时间依赖性(P<0.05),24 h、48 h的半数抑制浓度分别为67.81 μg/ml、45.76 μg/ml;DHA联合5-FU对细胞生长的抑制具有协同作用(CI<1,P<0.01),倒置显微镜下可见两药联合处理后细胞稀疏;亚二倍体峰比、Annexin-V早期凋亡率示DHA、5-FU均能诱导细胞凋亡且联合用药后细胞凋亡更明显(DHA、5-FU、联合组的亚二倍体峰比分别为5.2%、6.2%、13.9%;早期细胞凋亡率分别为4.00%、5.37%、13.11%);细胞周期曲线在联合组停滞于G0/G1和S期;RT-PCR显示DHA、5-FU可下调bcl-2和bcl 2l12表达,两药联合表达时下调更显著,bax表达则无明显改变.结论 DHA能抑制胃癌SGC 7901细胞增殖,联合5-FU后对细胞生长抑制和周期阻滞具有协同作用,可能通过下调bcl-2和bcl 2l12基因诱导胃癌SGC 7901细胞发生凋亡.     

Influence of p53 and bcl-2 on chemosensitivity in benign and malignant prostatic cell lines

The administration of cancer chemotherapeutic agents results in an increase in the apoptotic cells in the tumor: therefore, it has been assumed that anticancer drugs exhibit their cytotoxic effects via apoptotic signaling pathways. Characteristics that confer sensitivity to drug-induced apoptosis are, a functional p53 protein and expression of the apoptosis-promoting protein, bax. The role of p53 and bax/bcl-2 in drug-induced apoptosis was assessed in six prostate cell lines, 1532T, 1535T, 1542T, 1542N, BPH-1 and LNCaP using TD(50) concentrations of etoposide, vinblastine and estramustine. Cell death was monitored morphologically by fluorescent microscopy, and by flow cytometry (Annexin-V assay). Apoptotic morphology was rather low and ranged from 0.1% to 12.1%, 3.0% to 6.0% and 0.1% to 8.5% for etoposide, estramustine and vinblastine, respectively. Annexin-V binding and flow cytometry indicated apoptotic propensities of 0% to 4%, 0% to 3% and 0% to 5%, respectively. The percentage of cells responding to drug-induced apoptosis was, on average, higher in the tumor cell lines than in the normal cell lines, but showed no correlation with p53 status. The percentage of cells showing necrosis, assessed by Annexin binding and Propidium Iodide permeability in aqueous medium, tended to be much higher, and was found to be at the level of 5% to 30%. Immunoblotting demonstrated that bax and bcl-2 proteins were expressed at a basal level in all cell lines, but did not increase after exposure to TD(50) doses of the three drugs. The ratio of bax and bcl-2, measured by laser scanning densitometry, was not altered by the drug-induced DNA damage. The results suggest that apoptosis is not a major mechanism of drug-induced cell death in prostate cell lines and appears to be independent of p53 status and bax/bcl-2 expression.     

丁酸钠对人胰腺癌细胞ASPC-1生长的影响及其机制的研究

目的 探讨丁酸钠对人胰腺癌ASPC-1细胞生长的影响并探讨其作用机制.方法 使用不同浓度丁酸钠处理ASPC-1细胞.MTT法观察肿瘤细胞的增殖;流式细胞仪检测细胞凋亡和细胞周期的变化;Western印迹法检测细胞内p21、p53、bcl-2、bax蛋白表达的变化,实时定量PCR检测p21和bcl-2的表达.结果 丁酸钠能明显抑制ASPC-1细胞的增殖,其作用具有明显的时间和剂量依赖性.丁酸钠处理24 h后ASPC-1细胞凋亡率明显提高(P＜0.05),S期细胞比例显著降低(P＜0.05),G0/G1期细胞比例显著升高(P＜0.05).p21、bax蛋白表达明显上调(P＜0.05);bcl-2蛋白的表达显著降低(P＜0.05);p53蛋白表达无明显变化(P＞0.05).结论 丁酸钠可以通过诱导胰腺癌细胞的凋亡和周期阻滞而抑制癌细胞生长;其发生机制可能与下调抗凋亡基因bcl-2、上调促凋亡基因bax和肿瘤抑制基因p21的表达有关.

Abstract：

Objective To investigate the effects of sodium butyrate(NaBT) on proliferation of human pancreatic cancer cell line ASPC-1 and explore the possible mechanism. Methods The methylthiazolyl tetrazolium assay (MTT) method was used to detect cell proliferation and draw a curve. The cell apoptosis and cell cycle were determined with flow cytometry. Western blot was used to study the effect NaBT on the pancreatic cancer cells and explore its mechansim. Real-time PCR was employed to assess the expression levels of p53, p21, bcl-2 and cell cycle regulation gene p21. Results After incubation with different concentrations of NaBT for 24 to 72 h, ASPC-1 cell proliferation was inhibited dramatically. NaBT induced an increase of G0/G1 phase cells and a significant decrease in the ratio of S phase cells. The expression of p21 and bax was up-regulated at protein and mRNA level. The expression of bcl-2 was down-regulated at protein and mRNA level. There was no significant difference in the expression of p53 at protein and mRNA level. Conclusion TSA-induced growth inhibition is associated with a block in the G0/G1 phase and apoptosis, which may occur through down-regulating the expression of apoptosis gene bcl-2 and up-regulating the expression of cell cycle regulation gene p21and pro-apoptotic gene bax.     

Expression of bcl-2 homologue mRNAs in rat liver allograft: rejection-induced cell apoptosis is associated with upregulation of bax and bcl-xs expression

Abstract Apoptosis is considered to play an important role in rejection of organ transplants, although the precise mechanism has not been elucidated. In this study, we screened for the expression of bcl-2 homologues (bcl-2, bax, bcl-xl, and bcl-xs) and Fas ligand (FasL) by RT-PCR method in grafts during acute rejection in rats following liver transplantation. Both bax and bcl-xs (inducers of apoptosis) mRNA levels increased steadily in the allografted group from postoperative day (POD) 2 to 8, while no remarkable changes of bcl-2 and bcl-xl expression (inhibitors of apoptosis) were recognized. Significant induction of FasL gene expression was observed in the allografted group on POD 4 and expression gradually decreased thereafter, although minimal FasL mRNA expression was seen in isografts. Our results indicated, for the first time, that rejection-induced cell apoptosis is closely associated with upregulation of bax and bcl-xs expression besides FasL, but not with down-regulation of bcl-xl.     

Wild-type p53-dependent etoposide-induced apoptosis mediated by caspase-3 activation in human glioma cells

OBJECT: In an attempt to understand the roles of several apoptosis-related genes in human glioma cells, the authors investigated the relationship of wild-type p53, interleukin-1beta-converting enzyme (ICE), caspase-3 (CPP32), bax, and bcl-2 to the apoptotic response of three glioma cell lines after treatment with etoposide. METHODS: A human glioma cell line (U-87MG) that expresses wild-type p53, one that expresses mutant p53 (T-98G), and a T-98G derivative (T-98G/p53) that was transfected with a wild-type p53 expression vector (pCDM8-p53/neo) were used. Cell growth inhibition in response to etoposide was quantified using a modified methylthiazol tetrazolium colorimetric assay. Induction of apoptosis was evaluated using Hoechst 33258 staining and a DNA fragmentation assay. To study the expression of the apoptosis-related proteins and messenger RNAs in the three glioma cell lines, Western blotting and polymerase chain reaction were performed. A caspase assay and Western blot analysis were used to assess CPP32 and ICE protease activity. A CPP32 inhibition assay was used to determine whether a specific CPP32 inhibitor, DEVD-CHO, affects the apoptosis induced by etoposide in malignant glioma cells. Etoposide significantly inhibited the growth of U-87MG and T-98G/p53 cells in a dose-dependent manner compared with the growth of the T-98G cells. Treatment with low concentrations of etoposide resulted in the increased expression of wild-type p53; it also initiated CPP32 activity and induced apoptosis in the U-87MG cells. Apoptosis was not induced in T-98G cells at low concentrations of etoposide, although it was induced at high concentrations. Furthermore, low concentrations of etoposide also induced apoptosis in the T-98G/p53 cells by enhancing the expression of transfected wild-type p53, decreasing the expression of bcl-2, and activating CPP32 activity. However, etoposide did not alter the expression of bax and did not initiate ICE activity in these three glioma cell lines. Etoposide-induced apoptosis can be suppressed by the CPP32 inhibitor DEVD-CHO. CONCLUSIONS: These findings indicate that wild-type p53, CPP32, and bcl-2 may mediate apoptosis induced by etoposide. Forced expression of wild-type p53 increases etoposide cytotoxicity in human glioma cells by inducing apoptosis and may have important therapeutic implications.     

NDGA对HepG2细胞5-LOX及其凋亡相关基因表达的影响

目的 探讨脂氧合酶(lipoxygenase)抑制剂NDGA对肝癌细胞HepG2细胞5-LOX及其凋亡相关基因hTERT、bcl-2及bax表达的影响.方法 用RT-PCR方法检测NDGA对HepG2细胞5-LOX及其凋亡相关基因hTERT、bcl-2及bax表达的影响.结果 25、50、100、200 μmol/L的NDGA作用HepG2细胞24、48 h后5-LOX表达逐渐减弱,hTERTmRNA、bcl-2的表达明显下调,而bax的表达逐渐上调.与未加NDGA干预的HepG2对照组比较,P＜0.05.结论 5-LOX在HepG2细胞中存在高表达,提示5-LOX的高表达在肝细胞癌发病过程中可能起着重要作用;而端粒酶hTERTmRNA、bcl-2及bax等凋亡相关基因可能参与了其发病过程.脂氧合酶抑制剂NDGA体外可有效抑制细胞5-LOX的表达,明显下调端粒酶hTERTmRNA、bcl-2及上调bax的表达.LOX可能是肝癌生物化学治疗的新靶点.     

绿脓杆菌菌毛株对肝癌细胞Hep-2生长的抑制作用

目的 观察绿脓杆菌菌毛株菌苗(PA-MSHA)对人肝细胞癌细胞株HepG-2的抑制作用及机制.方法 噻唑蓝(MTT)法检测细胞生长抑制情况.原位末端标记(TUNEL)法检测细胞凋亡情况;透射电镜观察细胞超微结构.Western blot法检测bcl-2、bax、Caspase-3蛋白表达;裸鼠移植瘤内及瘤周药物注射观察不同药物浓度及作用时间的体内抑瘤作用.结果 肿瘤细胞的抑制率与药物浓度和作用时间成正比;高、低浓度实验组和对照组凋亡率差异有统计学意义(P＜0.01);电镜观察可见经PA-MSHA作用后的HepG2细胞呈现典型的凋亡表现;PA-MSHA作用后可引起bax、Caspase-3蛋白表达上调,bcl-2蛋白表达下调;高浓度PA-MSHA可明显抑制裸鼠移植瘤的生长,17 d抑瘤率为60%.结论 PA-MSHA可明显抑制HepG-2细胞的生长,诱导凋亡是其另外一个作用机制.     

苦参碱对增生性瘢痕成纤维细胞凋亡及相关调控蛋白表达的影响

目的　研究苦参碱 (matrine,Mat)对增生性瘢痕成纤维细胞 (fibroblast,FB)凋亡及相关调控蛋白表达的影响。方法　于 2 4只新西兰白兔耳部腹侧面制作增生性瘢痕模型。所有动物随机分为Mat组和对照组 (每组 12只 ) ,分别采用Mat(5 0 g/L)和等渗盐水于各组兔耳瘢痕处行局部封闭注射 ,1次 / 4d。用药 2、4、6、8、12周后 ,采用DNA切口末端标记技术 (TUNEL)检测各组增生性瘢痕FB的凋亡细胞数量 ,利用免疫组织化学方法检测凋亡相关调控蛋白 p5 3、bcl 2、bax的表达。 结果与对照组比较 ,用药 2、4、6、8、12周后 ,Mat组FB凋亡数量均明显增加 (P <0 .0 5 ) ,bax表达明显增强 ,p5 3、bcl 2表达明显减弱 ,瘢痕逐渐趋于平复。　结论　Mat可促进兔耳增生性瘢痕FB的凋亡 ,使凋亡相关调控蛋白bax表达上调 ,p5 3、bcl 2表达下调     

喜树碱诱导人膀胱癌细胞RT4和MGH凋亡的实验研究

目的 研究DNA拓扑异构酶Ⅰ抑制剂喜树碱(CPT)诱导人膀胱癌细胞凋亡的作用。方法 体外培养的膀胱癌RT4和MGH细胞用CFr处理，用FACScan流式细胞仪分析细胞凋亡的发生及变化，用Western blotting的方法检测了CFT处理不同时间段，bcl-2与bax表达的变化，以及天冬氨酸特异性半胱氨酸蛋白酶-3(Caspase-3)和多聚腺苷二磷酸核糖聚合物酶(PARP)的活性的改变。结果 CPT处理膀胱癌细胞24h后，细胞凋亡率明显上升，且CPT诱导的凋亡呈时间依赖性和剂量依赖性。CFT诱导膀胱癌细胞RT4和MGH后均未见bcl-2和bax表达水平的改变，但是可以观察到凋亡执行蛋白Caspase-3和PARP的激活。结论 CPT能够诱导膀胱癌细胞RT4和MGH发生凋亡，其抑制膀胱癌细胞的作用可能与肿瘤细胞发生凋亡有关。     

Apoptosis occurs after cerebral contusions in humans

OBJECTIVE: Animal model systems have shown that head trauma can induce cell death in regions of the brain away from the site of the impact via a process of apoptosis. We sought to determine whether there was evidence of cellular apoptosis in clinically collected materials from human head trauma patients, as well as to attempt to determine the pathway by which it may occur. METHODS: Thirty-one sequential specimens of brain tissue excised during emergency craniotomy for evacuation of cerebral contusions with mass effect were examined. Non-necrotic pericontusional tissues were detected in 11 samples. These were examined for the presence of apoptotic cells by the terminal deoxynucleotide transferase-mediated nick end labeling method as well as by immunohistochemistry to detect possible expression of the apoptosis-related genes p53, bcl-2, and bax. RESULTS: Bax expression was detected in all patients, whereas bcl-2 expression was noted in six patients. Terminal deoxynucleotide transferase-mediated nick end labeling-positive cells were noted in eight patients. One instance of p53-positive immunostaining was observed. Patients with bcl-2 expression had a better survival rate than patients in whom no bcl-2 expression was noted (P = 0.01). CONCLUSION: Although necrosis seemed to be the main finding in cerebral contusions, these results support the hypothesis that apoptosis does occur in patients after traumatic brain injury, and this may contribute to the secondary injury processes that are seen with head injury. Patients in whom anti-apoptotic bcl-2 is induced seem to have a better prognosis. This may have important clinical significance in the development of bcl-2 homologs or bax inhibitors to prevent apoptosis.