2-甲氧基雌二醇对SKM-1细胞凋亡中bcl-2/bax表达的影响

目的 探讨2-甲氧基雌二醇(2-ME)对骨髓增生异常综合征SKM-1细胞凋亡过程中bcl-2/bax表达的影响.方法 2-ME与SKM-1细胞共同培养,流式细胞术分析细胞凋亡率和细胞周期;RT-PCR技术检测bcl-2和bax mRNA表达.结果 1～8 μmol/L 2-ME作用后,SKM-1细胞凋亡率上升呈剂量依赖性;细胞被阻滞于G2/M期;细胞bcl-2 mRNA表达量下调,bax mRNA表达量上调.结论 2-ME诱导SKM-1细胞凋亡与细胞G2/M期阻滞和bcl-2/bax比率下降有关.     

15-脱氧前列素2抑制MG63细胞活性并诱导其凋亡的机制

目的研究过氧化物酶体增殖活化受体(PPAR)γ的内源性配体15-脱氧前列素2(15d-PGJ2)对骨肉瘤细胞株MG63的增殖活性的影响及诱导其凋亡可能的作用机制。方法采用人骨肉瘤细胞MG63,分别用终浓度为0.1、1、5、10和50μmol/L15d-PGJ2处理,对照组不给予药物,四甲基偶氮唑蓝(MTT)法测定药物作用24 h、48 h、72 h及96 h细胞增殖的变化;流式细胞术(FCM)结合Annexin V/PI双染色观察终浓度5μmol/L和20μmol/L15d-PGJ2作用48 h和72 h MG63细胞周期变化及诱导细胞凋亡和坏死情况;逆转录聚合酶链式反应(RT-PCR)检测终浓度20μmol/L15d-PGJ2处理48 h对MG63细胞中PPARγm RNA的表达影响;通过免疫细胞化学技术检测终浓度20μmol/L15d-PGJ2作用48 h,MG63细胞中凋亡相关蛋白caspase3,p53和Bax/bcl-2的表达情况。结果 MTT结果显示:1各浓度15 d-PGJ2作用24 h、48 h、72 h及96 h,对MG63细胞增殖均有抑制作用,48 h达到最佳抑制效果。48 h、72 h及96 h抑制率两两比较均无差异(P>0.05),且50μmol/L对细胞的抑制率几乎达100%;2在各时间点,各浓度组间比较均有差异(P=0.000),在0.1⁵0μmol/L浓度组区间,相同时间点的抑制率有随浓度升高而递增的趋势。流式细胞术检测结果显示:不同浓度的各细胞周期分布差异有统计学意义(P=0.000)。加药组G0/G1期细胞比例均高于对照组,S和G2/M期相应减少,且细胞凋亡率增高,表明加药组可使细胞G0/G1期阻滞且诱导细胞凋亡;RT-PCR检测PPARγm RNA的表达,并同时逆转录稳定的内参片段β-actin,以二者的面积灰度值比值作为该m RNA的相对表达量,20μmol/L 15d-PGJ2作用48 h,PPARγm RNA的相对表达量(2.06±0.35)与对照组(0.94±0.23)比较有差异(P<0.05);免疫细胞化学结果显示:药物干预组凋亡相关蛋白caspase3和Bax表达与对照组比较显著增加,而p53和bcl-2表达下降(P<0.05)。结论 1PPARγ激动剂15d-PGJ2能明显抑制人骨肉瘤MG63细胞的生长,引起细胞G1期阻滞并诱导细胞凋亡。215d-PGJ2可能通过增加PPARγm RNA表达,上调凋亡相关蛋白caspase3和Bax及下调p53和bcl-2表达而诱导细胞凋亡。     

IL-11促进胃癌细胞增殖的机制研究

目的研究IL-11对胃癌细胞增殖的促进作用及其分子机制。方法将培养的BGG823胃癌细胞株随机分为干预组和对照组,对照组使用不含药物的DMEM进行处理,干预组采用含有20μg/L、50μg/L rh IL-11的DMEM进行处理。处理后24 h、48 h时,PI染色后采用流式细胞仪测定细胞周期中G0/G1、S期、G2/M期细胞所占比例,抽提RNA后采用荧光定量PCR测定细胞中PCNA、Cyclin D1、CDK4、MMP2、MMP9的m RNA含量。结果处理24 h、48 h后,干预组细胞中G0/G1细胞的比例明显低于对照组,S期和G2/M期细胞的比例明显高于对照组(P0.05);50μg/L组细胞中G0/G1细胞的比例明显低于20μg/L组,S期和G2/M期细胞的比例显著高于20μg/L组(P0.05);干预组细胞中PCNA、Cyclin D1、CDK4的m RNA含量明显高于对照组(P0.05);50μg/L组细胞中PCNA、Cyclin D1、CDK4的m RNA含量明显高于20μg/L组(P0.05);干预组细胞中MMP2、MMP9的m RNA含量明显高于对照组(P0.05);50μg/L组细胞中MMP2、MMP9的m RNA含量明显高于20μg/L组(P0.05);且均与干预时间呈依赖性关系(P0.05)。结论 IL-11能够促进胃癌细胞的增殖以及细胞周期的发展,上调PCNA、Cyclin D1、CDK4、MMP2、MMP9的表达是IL-11发挥促增殖作用可能的分子机制。     

丹参酮ⅡA诱导PC-3细胞凋亡的细胞周期调控机制

目的研究丹参酮ⅡA诱导PC-3人前列腺癌细胞凋亡的细胞周期调控机制。方法 PC-3人前列腺癌细胞与不同浓度的丹参酮ⅡA共同培养48 h后,MTT法检测细胞增殖能力;流式细胞仪分析细胞周期;检测细胞周期调控因子CyclinD1和CDK4的表达。结果与对照组相比,0.25、0.5、1 mg/L丹参酮ⅡA组G0/G1期细胞数明显增多,S期细胞数明显减少(P<0.01);CyclinD1和CDK4的表达量均随丹参酮ⅡA剂量增加而减少(P<0.05,P<0.01)。结论丹参酮ⅡA拮抗PC-3人前列腺癌细胞增殖,其主要机制可能与CyclinD1及CDK4低表达所致细胞周期抑制有关。     

棕榈酸对大鼠胰腺腺泡细胞AR42J细胞凋亡的影响

目的探讨棕榈酸(palmitic acid,PA)对大鼠胰腺腺泡细胞AR42J的凋亡率的改变及凋亡相关基因bcl-2、bax、caspase-3mRNA表达的影响。方法 PA浓度分别为0 mmol/L(正常对照组)、0.1 mmol/L、0.5 mmol/L、1.0 mmol/L刺激AR42J细胞后,应用流式细胞术AV/PI双染法检测细胞的凋亡率,荧光定量PCR法检测凋亡相关基因bcl-2、bax、caspase-3 mRNA的表达水平。结果与正常对照组(0 mmol/L)相比,PA在0.1mmol/L低浓度时对AR42J细胞不具损伤作用,从0.5 mmol/L开始,PA对AR42J细胞的凋亡率呈浓度、时间依赖性增强,组间比较差异有统计学意义(P0.05)。荧光定量PCR结果显示PA下调抗凋亡基因bcl-2 mRNA的表达,同时上调促凋亡基因bax、caspase-3 mRNA的表达。结论不同浓度PA对AR42J细胞的损伤作用不同,主要表现为凋亡与剂量、时间相关,其机制可能是抑制bcl-2 mRNA的表达及促进bax、caspase-3 mRNA的表达。     

硫化氢在外源性SB203580预处理人卵巢癌细胞增殖与凋亡中的作用

目的探讨硫氢化钠(Na HS,H2S供体)通过p38丝裂原活化蛋白激酶(p38MAPK)信号通路对人卵巢癌细胞(SKOV3细胞株)增殖与凋亡的调控作用。方法体外培养人卵巢癌细胞,随机分为正常对照组、Na HS (0. 01 mol/L)组、SB203580(0. 1 mol/L)组、Na HS联合SB203580组,每组均经药物干预处理48 h后收集细胞,细胞增殖采用四甲基偶氮唑蓝(MTT)法检测,细胞周期分布与细胞凋亡率均采用流式细胞术检测。结果 (1)与正常对照组比较,Na HS组、SB203580组、Na HS联合SB203580组人卵巢癌SKOV3细胞抑制率均显著升高,G1期细胞比例均明显增加,S期细胞均明显减少,且细胞凋亡率均明显增加,差异有统计学意义(均P0. 01)。(2)SB203580组人卵巢癌SKOV3细胞抑制率、细胞周期各时相分布、细胞凋亡率均与Na HS组比较差异无统计学意义(P0. 05),而与Na HS组比较,Na HS联合SB203580组人卵巢癌SKOV3细胞抑制率明显增加,G1期细胞比例明显增加,S期及G2/M期细胞比例明显减少,且细胞凋亡率明显增高,差异有统计学意义(均P0. 01)。(3)与SB203580组相比,Na HS联合SB203580组人卵巢癌SKOV3细胞抑制率明显增加,G1期细胞比例明显增加,S期及G2/M期细胞比例明显减少,且细胞凋亡率显著增高,差异有统计学意义(均P0. 01)。结论 H2S可能通过抑制p38MAPK信号通路抑制人卵巢癌细胞的增殖,促进人卵巢癌细胞凋亡,且H2S可协同SB203580调节人卵巢癌细胞的增殖与凋亡。     

柠檬酸钠与三氧化二砷联合应用对胃癌SGC-7901细胞的作用

目的探讨柠檬酸钠(sodium citrate,CT)与三氧化二砷(As_2O_3,,AS)联合应用对胃癌SGC-7901细胞凋亡及作用机制。方法用sodium citrate(终浓度为5 mmol/L,CT5)和As_2O_3(终浓度依次为5 μmol/L,AS5)处理体外传代培养的胃癌SGC-7901细胞株。应用DAPI细胞核染色和流式细胞仪检测细胞凋亡和细胞周期的变化;RT-PCR法观察抗凋亡相关基因bcl-2表达的变化。结果 sodium citrate与As_2O_3联合应用相对于单用sodium citrate或As_2O_3可显著提高诱导胃癌细胞凋亡率(P0.05);与空白组相比,用药后G_0/G_1期细胞比例下降,G_2/M期细胞比例上升,细胞周期阻滞于G_2/M期,抗凋亡基因bcl-2表达明显下降。结论 sodium citrate与As_2O_3联合应用具有协同抗胃癌细胞的作用,其机制可能与细胞周期阻滞、增强诱导胃癌细胞凋亡以及调控凋亡相关基因的表达有关。     

60Coγ射线分割照射对宫颈癌细胞周期及相关蛋白的影响

目的 观察不同剂量60Coγ射线分割照射对宫颈癌Hela细胞周期和细胞周期素(cyclin) D1、B1及细胞周期蛋白依赖激酶(CDK)1、4的影响.方法 0、2、4、6 Gy 60Co γ射线分割照射宫颈癌Hela细胞,1次/d,共5 d.用流式细胞术检测细胞周期和凋亡指数,RT-PCR测定cyclin D1、B1及CDK1、4.结果 2、4、6 Gy 60Co γ射线分割照射后各照射组G0/G1和G2/M期细胞数量逐渐增多,细胞凋亡指数随照射剂量增加而增加,与对照组相比,P均<0.05;cyclin D1、CDK1在所有照射组中均不表达,而对照组细胞正常表达;各照射组细胞cyclin B1和CDK4的表达与对照组相比,P均<0.05. 结论 2、4、6 Gy 60Co γ射线分割照射后Hela细胞阻滞在G0/G1、G2/M期,并诱导细胞凋亡,照射后细胞周期相关蛋白表达均受到抑制.     

姜黄素对肝星状细胞增殖与凋亡的影响

目的　观察姜黄素对体外培养肝星状细胞 (HSC)增殖与凋亡的影响。方法　不同浓度姜黄素处理HSC株HSC T6 ,MTT法检测细胞增殖 ,流式细胞仪、透射电镜和琼脂糖凝胶电泳法检测细胞凋亡。结果　在 2 0～ 10 0 μmol/L浓度范围内 ,姜黄素可剂量依赖性地抑制HSC增殖 (P <0 .0 1)。 2 0、4 0、6 0 μmol/L姜黄素处理HSC 2 4h后 ,细胞周期分析发现S期细胞减少 ,G2 /M期细胞显著增加 (P <0 .0 1) ;流式细胞术检测到明显的亚G1峰 ,各组的凋亡指数 (% )分别是 15 .3± 1.9,2 6 .7± 2 .8,37.6± 4 .4 ,与对照组 (1.9± 0 .6 )相比 ,差异有显著性 (P <0 .0 1) ;4 0 μmol/L姜黄素作用 12、2 4、36、4 8h ,凋亡指数 (% )分别是 12 .0± 2 .4、2 6 .7± 3.5、33.8± 1.8和 4 9.3± 1.6 ,与对照组相比 ,差异有显著性 (P <0 .0 1) ;透射电镜观察到细胞皱缩、核染色质浓缩沿核膜排列和凋亡小体形成等 ;琼脂糖凝胶电泳可见到明显的DNA梯带形成。结论　姜黄素可显著抑制HSC增殖 ,使细胞周期停滞于G2 /M期 ,并诱导其凋亡 ,其作用具有时间和剂量依赖性     

熊果酸对食管癌KYSE105细胞增殖、凋亡的影响

目的 探讨熊果酸(ursolic acid,UA)对人食管癌KYSE105细胞增殖、细胞凋亡的影响及其分子机制.方法 应用0～50μmoL/L熊果酸处理KYSE105细胞24～72 h后,采用四甲基偶氮唑蓝法(MTT)检测细胞增殖抑制率;流式细胞术(FCM)检测KYSE105细胞周期及凋亡率;RT-PCR检测B细胞淋巴瘤/白血病-2(B cell lymphoma/lewkmia-2,bcl-2)、Bcl-2相关X蛋白(Bcl-2 as-sociated X protein,bax)及细胞周期素D1(cyclinD1)mRNA的表达.结果熊果酸作用后KYSE105细胞生长明显减慢,抑制率8.55%～89.48%(P<0.05),呈时间-剂量效应关系,使细胞周期阻滞在G0/G1期,凋亡率16.62%～21.54%(P<0.05).bax mR-NA表达升高,bcl-2、cyclinD1 mRNA表达下调(P<0.05).结论 熊果酸能明显抑制KYSE105细胞的增殖,诱导其凋亡,其机制可能与抑制bcl-2、cyclinD1表达,同时促进bax表达有关.     

血小板衍生生长因子在苯乙酸钠调控PC-3细胞增殖和凋亡中的作用

目的 探讨凋亡相关基因血小板衍生生长因子（PDGF）在苯乙酸钠（NaPA）调控人前列腺癌细胞株（PC-3）细胞增殖和凋亡过程中的作用.方法 在PC-3细胞培养液中加入不同浓度的NaPA,采用流式细胞术（FCM）检测PC-3细胞增殖和凋亡情况,同时用RT-PCR检测PDGFmRNA表达.结果 在体外经不同剂量NaPA处理后的PC-3细胞,随着NaPA浓度的增加则增殖指数（PI）逐渐降低（P＜0.001）;凋亡细胞数逐渐增多（P＜0.001）;在前列腺癌PC-3细胞凋亡过程中,PDGFmRNA水平随NaPA剂量的增加,表达量降低（P＜0.001）.结论 提示PDGF基因可能在由NaPA调控的前列腺癌细胞系PC-3细胞增殖和凋亡过程中起着重要作用.     

瘦素对人胆管癌细胞QBC939增殖和凋亡的影响

目的观察瘦素对人胆管癌细胞QBC939增殖和凋亡的影响并初步探讨其可能的机制。方法在培养液中加入不同浓度的瘦素后,用四甲基偶氮唑盐（MTT）法检测细胞增殖,流式细胞仪观察细胞周期及凋亡情况,实时荧光定量PCR法检测增殖相关基因Cyclin D1和凋亡相关基因Bax、Bcl-2的表达情况,同时检测Caspase-3的活性。结果 MTT法显示瘦素可以促进QBC939细胞的增殖;流式细胞仪检测结果显示瘦素能明显降低G0/G1期细胞比例并提高S期细胞比例,细胞凋亡率也有明显降低;实时荧光定量PCR法显示瘦素（50 ng/ml）处理QBC939细胞24 h后,Cyclin D1mRNA的表达明显增高,Bcl-2 mRNA的表达量明显增高,而Bax mRNA的表达量下降,差异有统计学意义;而且瘦素作用QBC939细胞后能降低细胞的Caspase-3酶活性。结论瘦素可以明显的促进人胆管癌细胞QBC939从细胞周期G0/G1期向S期转换,进而促进细胞增殖,此外瘦素还可以抑制其凋亡。     

Interferon tau-induced hepatocyte apoptosis in sheep

Clinical applications of Type I interferon (IFN) are limited by adverse side effects mediated largely by unknown mechanisms. This study examined the mechanisms of acute hepatic injury in lambs treated with systemic administration of IFN-tau, a Type I IFN. Liver tissues were collected at 24, 48, or 96 hours after treatment with either IFN-tau or saline. Histopathology revealed acute hepatopathy including cellular swelling, cytoplasmic aggregates, and apoptosis in all IFN-tau-treated lambs, which were accompanied by elevation of aspartate transaminase (AST) (P <.01). The number of apoptotic hepatocytes in IFN-tau-treated lambs was higher than for control lambs (P <.001). Immunohistochemistry for proliferating cell nuclear antigen (PCNA) revealed that IFN-tau induced hepatocyte growth arrest at the G0/G1 phase of the cell cycle and that the majority of hepatocytes in S or G2 phase were eliminated by apoptosis. We investigated expression of bax-alpha and bcl-2, acting as pro- and antiapoptotic molecules, in IFN-tau-induced apoptosis. Northern blot analysis revealed increased expression of bax-alpha messenger RNA (mRNA) in IFN-tau-treated lambs (P <.01) compared with control lambs, consistent with the expression pattern for bax-alpha protein. However, there was no detectable difference in expression of bcl-2 proteins between control and IFN-tau-treated lambs. The levels of bax-alpha associated with the mitochondria also increased during IFN-tau treatment. Bax-alpha immunostaining showed scattered immunoreactive hepatocytes with morphological hallmarks of apoptosis. These results suggest that IFN-tau induces growth arrest as well as apoptosis by regulating bax-alpha expression. These pathological effects of IFN-tau on sheep liver indicate potential mechanisms of Type 1 IFN-induced hepatotoxicity in animals and humans.     

Mechanism of apoptotic effects induced selectively by ursodeoxycholic acid on human hepatoma cell lines

AIM: To investigate the effects of ursodeoxycholic acid (UDCA) on apoptosis and proliferation of hepatoma cell lines. METHODS: Human hepatoma cell lines HepG2 and BEL 7402 were cultured in medium supplemented with different concentrations of UDCA, normal human hepatic line L-02 was used as control. Cell proliferation, apoptosis and gene expression were detected using methyl thiazolyl tetrazolium (MTT) assay, flow cytometry, Western blot, DNA ladder assay, electron microscopy, and immunocytochemistry. RESULTS: Ursodeoxycholic acid inhibited the proli- feration of HepG2 and BEL7402 cell lines in a dose- dependent manner. Ursodeoxycholic acid can change cell cycle distribution of HepG2 and BEL7402, the proportion of cells in G0-G1 phase increased whereas the proportion of S phase cells and G2-M phase cells decreased. Ursodeoxycholic acid arrested the cell cycle in G0-G1 phase by down-regulating the cell cycle related proteins cyclin D1, D3 and retinoblastoma protein (pRb). The apoptotic rates of HepG2 and BEL7402 treated with UDCA (1.0 mmol/L) were significantly higher than those of control. In the HepG2 and BEL7402 treated with UDCA, expression of bcl-2 decreased whereas expression of Bax increased, the nuclear fragmentation and chromosomal condensed, cells shrank and lost attachment, apoptotic bodies and DNA ladders appeared. UDCA had no effect in inducing apoptosis on L-02 cell lines. CONCLUSION: UDCA can selectively inhibit proliferation and induce apoptosis of HepG2 and BEL7402 cell lines by blocking cell cycle and regulating the expression of Bax/bcl-2 genes.     

不同用药顺序吉非替尼、多西他赛联用对非小细胞肺癌细胞的体外抑制作用

目的探讨不同用药顺序吉非替尼、多西他赛(DXT)联用对非小细胞肺癌(NSCLC)细胞的体外抑制作用。方法选择人肺癌细胞株PC-9、PC-9/G、A549,用MTT法、流式细胞技术分别检测不同顺序吉非替尼、DXT联用对3种细胞存活率、周期及凋亡的影响;用药顺序分先用DXT、后用吉非替尼(D→G组)及先用吉非替尼、后用DXT组(G→D组)。结果与G→D组比较,D→G组细胞存活率显著减少,可使细胞阻滞在G2～M期,并增强DXT诱导的细胞凋亡(P均<0.05);G→D组可使细胞阻滞在G0～G1期,并减少DXT诱导的细胞凋亡。结论先用DXT、后用吉非替尼为抑制NSCLC细胞增殖的最佳联用方案。     

Effect of arsenic trioxide on rat hepatocarcinoma and its renal cytotoxicity

AIM: To study the effect of arsenic trioxide (As2O3) on rat experimental hepatocarcinoma and its renal cytotoxicity.METHODS: The hepatocarcinoma model was established by diethaylnitrosamine perfusion in stomach of 120 Wistar rats, and the treatment began at the end of 20 weeks.Before the treatment, the rat models were randomly divided into 5 groups. In the treatment groups, three doses of As2O3 were injected into rat abdominal cavity, the total time of drug administration was 4 weeks. Cisplatin control or the blank group was injected into abdominal cavity with equal amount of cisplatin or saline at the same time,respectively. On the 7th, 14th and 28th day after the treatment, the hepatocarcinoma nodules were obtained and the morphologic changes of hepatocarcinoma cells were observed under light and electron microscopes;Immunohistochemistry (S-P methods) was employed to detect the expression of bcl-2, bax and PCNA in hepatocarcinoma tissues; flow cytometry (TUNEL assay)was used to detect the apoptosis of liver cancer cells and the change of cytokinetics. On the 28th day, the kidneys were obtained and their histologic changes were observed under light microscope, and immunohistochemistry (SP stain) was also employed to detect the expression of bcl-2and PCNA. Cisplatin and saline solution were used as the control.RESULTS: As2O3 could induce the apoptosis of rat liver cancer cells and exhibited typical morphologic changes.The incidence of apoptosis of hapatocarcinoma cells was elevated (P=0.001). The elevation was the most higher in the group of middle-dose of As2O3 (1 mg.kg-1), significantly higher than that of the other arsenic groups and the controls (P=0.001). Large dose of As2O3 (5 mg.kg-1) was able to arise the incidence of apoptosis, but also produced a large amount of necrosis and inflammatory reaction. Middle dose of As2O3 dramatically increased the cell number in G2/M phase (P=0.0001), and apoptosis happened apparently.The expression of bcl-2 and bax was related to the dose of As2O3. With the up-regulation of apoptotic incidence, the ratio of bcl-2/bak decreased. But the incidence of apoptosis was not the highest status and the ratio of bcl-2/bax was at the lowest when the highest-dose of As2O3 was used.There was significant difference among the PCNA indexes (PCNA L1) of the five groups. Of them, three arsenic groups all showed decrease of different degrees, and this downregulation was most obvious in group A. There was significant difference among the three groups (P=0.016).Under the light microscope, the rat kidney in the cisplatin group exhibited tubular epithelium swelling and degeneration, protein casts in collecting tubules; While all arsenic groups didn't show the significant changes (P=0.013).In the arsenic groups, the expression of bcl-2 in the renal tubular epithelium was increased (P=0.005), no obvious changes happened to PCNA L1. But in the group of cisplatin,the PCNA L1 increased significantly (P=0.001).CONCLUSION: AS2O3 can induce apoptosis of rat hepatocellular carcinoma cells. And there is optimum dose;too high dose will induce the cytotoxic effect, while certain dose of As2O3 is able to block the cell cycle at G2/M phase.As2O3 had the most remarkable influence on G2/M cells,and it can also induce apoptosis to cells at other phases.As2O3 can restrain the proliferation of rat hepatocellular carcinoma cells, in a dose-time dependent manner.Compared with cisplatin, As2O3 didn't show obvious renal toxicity, which was related to the increasing expression of bcl-2 in renal tubular epithelium, the inhibition of apoptosis and the anti-oxidation effects.     

tBHQ对无机砷致HaCaT细胞细胞周期调节失控的拮抗作用

目的探讨叔丁基对苯二酚(tert-butylhydroquinone,tBHQ)对亚砷酸钠(sodium arsenite,NaAsO2)致人皮肤角质细胞系HaCaT细胞周期阻滞及其调控蛋白异常改变的拮抗作用。方法流式细胞仪法测定细胞周期;Western Blot检测细胞内Cyclin D1和CDK4的蛋白表达情况。结果 NaAsO2单独作用HaCaT细胞,G0/G1期细胞比例明显下降(P<0.01),S期和G2/M期细胞比例较对照组明显增加(P<0.01);tBHQ预处理后,tBHQ(50μmol/L)预处理的NaAsO2(25、50μmol/L)组细胞G0/G1期细胞比例有所上升(P<0.01);tBHQ(50μmol/L)预处理的NaAsO2(50μmol/L)组S期细胞比例明显下降(P<0.01);G2/M期细胞比例整体呈下降趋势。NaAsO2单独作用于HaCaT细胞,细胞周期相关蛋白Cyclin D1和CDK4的蛋白表达随NaAsO2染毒浓度的增加而明显下降(P<0.01);tBHQ预处理后,Cyclin D1和CDK4蛋白表达均明显高于相同浓度NaAsO2单独作用组(P<0.01)。结论 tBHQ对无机砷致人皮肤角质细胞细胞周期异常变化具有一定的拮抗作用。     

蛋白酶的活化和bcl-2基因的表达对家兔缺血再灌注心肌细胞凋亡的影响

目的探讨半胱氨酸天冬氨酸蛋白酶-3的活化和bcl-2基因的动态表达对家兔缺血再灌注心肌细胞凋亡的影响。方法42只家兔建立心肌缺血再灌注模型,随机分为对照组(6只)和实验组(36只)。采用原位杂交、免疫组织化学技术等,观察家免缺血再灌注后半胱氨酸天冬氨酸蛋白酶-3 mRNA、bcl-2 mRNA动态表达的变化规律对心肌细胞凋亡细胞的数目影响。结果缺血再灌注损伤激发了半胱氨酸天冬氨酸蛋白酶-3的活化,促进了凋亡的发生,二者的曲线变化呈正相关(r=0．9286,P<0．001)。半胱氨酸天冬氨酸蛋白酶-3、bcl-2之间形成一种抗衡的网络调控构象,负责凋亡的促进和凋亡的抑制。半胱氨酸天冬氨酸蛋白酶-3 mRNA、bcl-2mRNA、细胞凋亡三者的表达,在交界区比梗死区更明显。结论半胱氨酸天冬氨酸蛋白酶-3 mRNA活化诱导的细胞凋亡是家兔缺血再灌注引起心肌细胞损伤的主要原因。交界区以凋亡为主要变化的动态演变,为临床治疗提供了一个时间窗。     

As2O3对肾癌细胞增殖、凋亡的影响及意义

目的观察As2O3对肾癌细胞株786-0增殖、凋亡及细胞内X染色体连锁凋亡抑制蛋白（XIAP）表达的影响,探讨其意义。方法取对数生长期786-0,分别经0．5、1．0、2．0μmol／L的As2O3处理后,采用MTT比色法检测细胞生长情况,流式细胞仪测算细胞凋亡率,蛋白质印迹法及RT—PCR法检测细胞中的XIAP及其mRNA。结果0．5、1．0、2．0μmol／LAs2O3均可抑制786-0增殖。随着作用时间的延长,细胞生长抑制率升高（P均〈0．01）;随着As2O3浓度的增加,细胞凋亡率升高（P均〈0．01）、细胞内XIAP及其mRNA表达明显下调（P均〈0．05）。结论As2O3可抑制786-0增殖,并诱导其凋亡。这一作用与As2O3抑制786-0中XIAP及其mRNA表达有关。     

棓丙酯对高糖诱导人肾小管上皮细胞损伤的保护作用

目的 观察樯丙酯对高糖诱导人肾小管上皮细胞损伤的保护作用.方法 传代培养人肾小管上皮细胞（HK-2）,分为对照组、高渗组、高糖组、处理组1（桔丙酯浓度2μg/ml）和处理组2（棓丙酯浓度4μg/ml）,观察各组细胞增殖能力、凋亡率、活性氧（ROS）水平以及bcl-2和BaxmRNA的表达情况.结果 高糖刺激HK-2细胞后,细胞增殖能力下降、凋亡增加、ROS水平升高、bcl-2 mRNA表达减少、Bax mRNA表达增多.棓丙酯可以提高细胞增殖能力,减少凋亡,降低ROS水平,上调bcl-2 mRNA表达以及下调BaxmRNA表达;处理组2上述改变较处理组1明显（P均＜0.05）.结论 桔丙酯改善高糖诱导的人肾小管上皮细胞损伤,其机制可能与减轻氧化应激、上调bcl-2 mRNA表达以及下调Bax mRNA表达有关.