单核细胞增多型李氏忒菌诱导小鼠胸腺细胞凋亡

以流式细胞仪 (FCM)分析 ,DNA琼脂糖凝胶电泳分析及细胞形态学改变为指标 ,研究单核细胞增多型李氏忒菌 (LM)感染对小鼠胸腺细胞凋亡的诱导作用。结果发现 ,LM能诱导小鼠胸腺细胞产生典型的细胞凋亡形态学改变 ;FCM分析显示特征性的凋亡峰 ;琼脂糖凝胶电泳分析显示胸腺细胞出现典型的DNA“梯状带” ;胸腺细胞凋亡于LM(5× 10 5CFU)感染后 8h出现 ,48h达高峰 ;胸腺萎缩于LM感染后 16h出现 ,48h达最低水平 ;胸腺细胞凋亡百分率随LM感染剂量增加而增高。提示 ,LM以时间和剂量依赖方式诱导小鼠胸腺细胞凋亡。     

七味白术散对人轮状病毒感染乳鼠胸腺细胞的程序性死亡和IL-2、IL-2R表达的影响

观察七味白术散对人轮状病毒（HRV）感染乳鼠胸腺细胞的程序性死亡和IL－2、IL－2R表达的影响。NIH乳鼠经口感染HRV建立感染模型,采用DNA凝胶电泳、细胞形态学、ELISA方法观察处理对HRV感染乳鼠胸腺细胞凋亡以及IL－2、IL－2R表达的影响。七味白术散100g/L可明显抑制乳鼠胸腺细胞,药物干预10h未出现明显的细胞凋亡梯带,处理各组胸腺细胞24h均可见典型凋亡形态和DNA梯状带,而新鲜的胸腺细胞则无。七味白术散治疗组乳鼠血清中IL－2、IL－2R表达上调。七味白术散能延缓HRV感染乳鼠胸腺细胞凋亡,促进IL－2、IL－2R的表达,显示出七味白术散一种新的免疫调节效应。     

HSF1抑制热应激所致RAW264.7巨噬细胞凋亡

目的:探讨热休克因子1(heat shock factor 1,HSF1)对热应激所致Raw2 6 4.7巨噬细胞凋亡的影响。方法:采用热应激(4 2.5℃±0.5℃)处理稳定表达小鼠HSF1基因的Raw2 6 4.7巨噬细胞1h,3 7℃分别恢复6,9,1 2,2 4 h,采用流式细胞术,hoechst3 3 2 5 8染色和DNA琼脂糖凝胶电泳检测细胞凋亡。结果:流式细胞术结果显示,热应激后对照组(转空载体)细胞凋亡核百分率较热应激前明显升高,9 h达峰值(约为6 0%),此时荧光染色可见3 0%的细胞出现核固缩,凋亡小体等典型的凋亡形态学改变;并于热应激后6,9,1 2 h均能检测到清晰的DNA梯状条带。与转空载体对照组相比,HSF1过表达能显著降低热应激所致凋亡及明显抑制DNA的断裂。结论:HSF1可以抑制热应激所致的Raw2 6 4.7巨噬细胞凋亡。     

沙眼衣原体感染细胞抗凋亡及Bim的表达

目的:研究沙眼衣原体(D血清型)感染的Hela229细胞Bim蛋白质的表达及抵抗凋亡的情况。方法:Western-blot检测沙眼衣原体感染和未感染的Hela229细胞Bim蛋白质的表达水平。凋亡诱导剂etoposide作用Hela229细胞后,琼脂糖凝胶电泳检测DNA Ladder带;流式细胞仪检测凋亡率。结果:Hela229细胞在未感染沙眼衣原体时可检测到Bim的表达;在感染24小时、48小时后均未检测到Bim的表达。经etoposide作用后,未感染的Hela229细胞检测到DNALadder带;流式细胞仪检测的凋亡率为90.64%。感染24小时的Hela229细胞,未检测到DNA Ladder带;凋亡率为11.50%,与未感染的Hela229细胞诱导后的凋亡率比较有统计学意义(P<0.05)。结论:沙眼衣原体感染的Hela229细胞Bim蛋白质的表达下降;并能抵抗etoposide诱导的细胞凋亡。     

斑蝥酸钠体外诱导肝癌细胞凋亡的实验研究

目的 :探讨斑蝥酸钠体外诱导肝癌细胞的凋亡。方法 :应用MTT法、流式细胞仪、透射电镜和琼脂糖凝胶电泳综合分析肝癌细胞的变化。结果 :斑蝥酸钠作用 4 8h使肝癌细胞生长明显抑制 ;细胞凋亡率达 4 0 .0 2 %±2 .0 5 % ,并出现典型的凋亡峰 ,细胞增殖于G2 +M期阻滞 ;细胞核染色质出现凝集、边集等超微结构变化 ;细胞DNA电泳也显示了凋亡特有的“梯子”状条带。结论 :斑蝥酸钠体外可诱导肝癌细胞的凋亡 ,细胞生长阻断在G2 +M期。     

人参总皂苷诱导HL-60细胞凋亡作用及对Caspase-3、Fas表达的影响

目的 探讨人参总皂苷诱导HL-60细胞凋亡的作用机制及其对凋亡相关基因Caspase-3、Fas表达的影响.方法 采用人参总皂苷0、100、200及400 μ g/ml作用于HL-60细胞,并在不同时间内用MTT法检测人参总皂苷对HL-60细胞的抑制率;48h后予普通光镜、荧光显微镜检测HL-60细胞形态的变化;予流式细胞仪、DNA凝胶电泳检测HL-60细胞凋亡;予RT-PCR法检测Caspase-3、Fas基因表达的变化.结果 人参总皂苷抑制HL-60细胞生长呈剂量及时间依赖性;在100～400 μ g/ml时,HL-60细胞凋亡逐渐增加,可见典型的凋亡细胞形态和明显的DNA梯度;在该浓度范围内,Caspase-3、Fas表达逐渐增加.结论 一定浓度的人参总皂苷可诱导HL-60细胞发生凋亡,且细胞凋亡率与人参总皂苷浓度呈一定的量效依赖关系,Caspase-3、Fas基因的表达变化在人参总皂苷诱导HL-60细胞凋亡中可能起重要作用.     

Induction of SiHa Cells Cells Apoptosis by Nanometer Realgar Suspension and Its Mechanism

The effects of nanometer realgar suspension on proliferation and apoptosis of human uterine cervix cancer cell line SiHa cells and oncogenic genes HPV16E6/E7 were investigated. A "micro-jet efflux" strategy was used for the preparation of nanometer realgar suspension. SiHa cells were treated with nanometer Realgar suspension in various concentrations (6.25,12.5,25 and 50mg/L) for different durations (12,24,48 and 72h). The inhibitive effect of nanometer realgar suspension on growth of SiHa cells was detected by MIT method. Special morphological changes of apoptosis were observed by transmission electron microscopy (TEM) and DNA fragments electrophoresis. The apoptotic rate was quantified by flow cytometry (FCM). The expression of HPV16E6/E7 mRNA and protein was assayed by RT-PCR and Western blot respectively. The results showed after being treated with 25-50mg/L nanometer realgar suspension for 48h, the survival rate of SiHa cells was decreased, and apoptotic rate markedly increased in a time- and concentration-dependent manner. TEM and DNA electrophoresis revealed the special morphological changes of apoptosis. The apoptotic rate of SiHa cells treated with nanometer realgar suspension was significantly higher than in the control group (P<0.01), and G0/G1 phase arrest appeared following treatment with nanometer realgar suspension in 25 and 50mg/L for 48h.RT-PCR and Western blot assay indicated that nanometer realgar suspension reduced the HPV16E6/E7 gene expression. Nanometer realgar suspension could inhibit the proliferation and induce apoptosis of SiHa cells. The mechanism may be related to the down-regulation of the HPV16E6/E7 gene expression.     

模拟缺血/再灌注诱导人离体胃黏膜上皮细胞的凋亡

目的模拟缺血/再灌注诱导人离体的胃黏膜上皮细胞损伤模型,观察再灌注不同时间点胃黏膜上皮细胞凋亡情况并分析其规律。方法采用人胃黏膜上皮细胞株进行体外培养并传代,细胞以缺氧（5%CO2、1%O2、94%N2）＋无糖KR液模拟缺血、缺氧,复氧、复糖模拟再灌注诱导胃黏膜上皮细胞凋亡,用流式细胞仪、Ho-echst33258染色法观察正常对照组及模拟损伤组于模拟缺血1 h,再灌注3、6、12 h细胞凋亡的情况及变化规律。结果①胃黏膜上皮细胞12～24 h贴壁,24 h后有少量上皮样细胞从周边爬出,48 h后细胞呈指数样生长,72 h后细胞已基本能达到融合。②模拟胃缺血/再灌注后细胞凋亡率于缺血1 h、再灌注6 h时出现高峰,且与正常对照组相比差异有统计学意义（P〈0.01）。经Hoechst33258染色,荧光显微镜下观察可见细胞于缺血1 h、再灌注6h时产生典型的凋亡形态学变化如核浓染、核碎裂、边集等。结论成功模拟了缺血/再灌注诱导人离体的胃黏膜上皮细胞损伤模型;该损伤模型可引起胃黏膜上皮细胞的凋亡,且在缺血1 h、再灌注6 h时为高峰。     

Induction of SiHa Cells Apoptosis by Nanometer Realgar Suspension and Its Mechanism

The effects of nanometer realgar uterine cervix cancer cell line SiHa cells and suspension on proliferation and apoptosis of human oncogenic genes HPV16E6/E7 were investigated. A ＂micro-jet effiux＂ strategy was used for the preparation of nanometer realgar suspension. SiHa cells were treated with nanometer Realgar suspension in various concentrations （6.25, 12.5, 25 and 50 mg/L） for different durations （12, 24, 48 and 72 h）. The inhibitive effect of nanometer realgar suspension on growth of SiHa cells was detected by MTT method. Special morphological changes of apoptosis were observed by transmission electron microscopy （TEM） and DNA fragments electrophoresis. The apoptotic rate was quantified by flow cytometry （FCM）. The expression of HPV 16E6/E7 mRNA and protein was assayed by RT-PCR and Western blot respectively. The results showed after being treated with 25--50 mg/L nanometer realgar suspension for 48 h, the survival rate of SiHa cells was decreased, and apoptotic rate markedly increased in a time- and concentration-dependent manner. TEM and DNA electrophoresis revealed the special morphological changes of apoptosis. The apoptotic rate of SiHa cells treated with nanometer realgar suspension was significantly higher than in the control group （P〈0.01）, and G0/G1 phase arrest appeared following treatment with nanometer realgar suspension in 25 and 50 mg/L for 48 h. RT-PCR and Western blot assay indicated that nanometer realgar suspension reduced the HPV16E6/E7 gene expression. Nanometer realgar suspension could inhibit the proliferation and induce apoptosis of SiHa cells. The mechanism may be related to the down-regulation of the HPV16E6/E7 gene expression.