目的 利用分析各种浓度环氧化酶-2(COX-2)特异度抑制剂塞来昔布对食管癌EC109细胞系的作用,进而对COX-2蛋白表达的影响及对细胞凋亡能力的作用,进一步探讨塞来昔布对食管癌细胞凋亡的作用及机制。方法 使用0 μmol/L、20 μmol/L、60 μmol/L、100 μmol/L四个浓度的塞来昔布处理EC109细胞24 h,酶联免疫吸附剂测定(ELISA)法测定COX-2蛋白表达;流式细胞仪测定EC109细胞凋亡情况。结果 与0 μmol/L塞来昔布组比较,20 μmol/L、60 μmol/L、100 μmol/L塞来昔布组EC109细胞内COX-2蛋白表达不断降低(1.581±0.116;1.226±0.089,0.846±0.076,0.521±0.082)(P<0.05);而细胞凋亡率逐步上升(1.700±0.557,13.400±1.735,18.766±1.301,28.100±1.997)(P<0.05)药物浓度依赖于梯度。结论 塞来昔布是一种COX-2抑制剂,可能以浓度梯度的形式抑制COX-2蛋白的表达,从而促进EC109细胞的凋亡。

Objective The effects of celecoxib, a specific COX-2 inhibitor at various concentrations, on EC109 cell line of esophageal cancer were analyzed, and the effect and mechanism of celecoxib on apoptosis of esophageal carcinoma cells were further studied. Methods EC109 cells were treated with celecoxib at concentrations of 0 μmol/L, 20 μmol/L, 60 μmol/L and 100 μmol/L for 24 h. The protein of COX-2 in EC109 cells was determined by enzyme-linked immunosorbent assay (ELISA). Assay of EC109 cell apoptosis were determined by flow cytometry. Results Compared with the 0μmol/L celecoxib group, the expression of COX-2 protein in EC109 cells of 20μmol/L, 60μmol/L, 100μmol/L celecoxib group gradually decreased(1.581±0.116; 1.226±0.089, 0.846± 0.076, 0.521±0.082) (P<0.05); and the apoptotic rate gradually increased (1.700±0.557; 13.400±1.735, 18.766±1.301, 28.100±1.997) (P<0.05) in a drug concentration gradient-dependent manner. Conclusion The COX-2 inhibitor celecoxib may inhibit the expression of COX-2 protein in a concentration gradient and promote the apoptosis of esophageal cancer EC109 cells.

目的 在原来研究的基础上进一步研究Wnt-1信号通路蛋白-3（WISP-3）在高氧诱导肺上皮细胞凋亡中的作用。方法 通过Western blot检测和免疫组化检测不同肺上皮细胞中WISP-3的蛋白表达量。利用质粒转染和siRNA的方法在Beas-2B细胞中高表达和基因沉默WISP-3,通过细胞活性检测和流式细胞学技术检测高氧刺激后细胞的凋亡情况。结果 与空气对照相比,高氧刺激使肺上皮细胞的WISP-3蛋白表达量下降;WISP-3基因沉默或高表达使高氧诱导的肺上皮细胞凋亡增加或减少。结论 高氧刺激下,肺上皮细胞中WISP-3表达下降,WISP-3对高氧诱导的肺上皮细胞凋亡具有保护作用。

Objective To explore how Wnt-1 inducible signaling pathway protein-3 （WISP-3） participate in and play a regulatory role in the process of hyperoxia induced apoptosis in lung epithelial cells. Methods The expression of WISP-3 was detected via Western blot and immunohistochemistry. High expression and low expression of WISP-3 were performed by plasmid transfection and siRNA. Cell viability and flow cytometry were executed to detect the hyperoxia-induced apoptosis in Beas-2B. Results Compared to the group of air control,the expression of WISP-3 protein in lung epithelial cells decreased obviously after hyperoxia. Cell survival decrease and apoptosis increased after hyperoxia in Beas-2B cells with low expression of WISP-3. Vice versa. Conclusion The expression of WISP-3 decreased after hyperoxia in lung epithelial cells. The role of WISP-3 in this process may be protective.

目的 氧化苦参碱对视网膜母细胞瘤细胞SM-106凋亡的诱导作用及机制。方法 以不同作用时间(24 h、48 h、72 h)和不同作用浓度(12.5 μl/mL、25 μl/mL、50 μl/mL、100 μl/mL)氧化苦参碱处理视网膜母细胞瘤细胞SM-106,分别采用流式细胞仪及western blot检测视网膜母细胞瘤细胞SM-106细胞凋亡及其凋亡因子(Bax、Bcl-2)蛋白表达。结果 氧化苦参碱可促进SM-106细胞体外凋亡,上调Bax蛋白表达及Bax/Bcl-2蛋白表达比值,下调Bcl-2蛋白表达,并呈现剂量及时间依赖性。结论 氧化苦参碱可诱导视网膜母细胞瘤细胞SM-106凋亡,调控凋亡因子Bax、Bcl-2的表达是其可能作用机制。

Objective To evaluate the apoptosis and its mechanism of retinoblastoma cells SM-106 induced by oxymatrine. Methods Retinoblastoma cells SM-106 were treated with different time(24 h、48 h、72 h)and different concentrations(12.5 μl/mL, 25 μl/mL, 50 μl/mL or 100 μl/mL) of oxymatrine. The apoptosis and protein expression of apoptosis factors (Bax and Bcl-2) were respectively determined by flow cytometry and western blot. Results Oxymatrine significantly promoted the SM-106 cells apoptosis in vitro, raised Bax protein expression and Bax/Bcl-2 protein expression ratio, reduced the Bcl-2 protein expression, and showed the dose and time dependent. Conclusion Oxymatrine is able to induce the apoptosis in retinoblastoma cells SM-106. Regulating apoptosis related gene Bax and Bcl-2 expression may be the mechanism of apoptosis.

目的 观察葛根素对新西兰白兔视网膜缺血/再灌注损伤组织中细胞凋亡的保护作用。方法 新西兰白兔30只随机分为缺血再灌注对照组和葛根素治疗实验组,各组右眼应用前房灌注加压法建立视网膜缺血再灌注模型,分别于再灌注后第12、24、72 h处死动物,摘除眼球,制作石蜡切片,用TUNEL法检测细胞凋亡,计算凋亡指数。结果 对照组缺血再灌注12 h在神经节细胞层和内核层可见凋亡细胞;24 h神经节细胞层细胞数有所减少,视网膜神经节细胞层、内核层及外核层均见凋亡细胞明显增多;72 h神经节细胞层细胞数明显减少,神经节细胞层、内核层及外核层仍见凋亡细胞,但较24 h有所减少。葛根素治疗视网膜的凋亡细胞在各个时间段的表达规律与对照组相似,但凋亡细胞计数在12 h,24 h,72 h均较对照组明显减少,两组间差异有统计学意义。结论 葛根素能减轻缺血-再灌注损伤的视网膜细胞凋亡,对视网膜有保护作用。

Objective To observe the protective effects of Puerarin on apoptosis of ischemic injury in rabbit retina. Methods Retinal ischemia was induced in rabbits by increasing intraocular pressure to 120 mmHg for 60 minutes. TdT-mediated biotin-dUTP nick end labelling(TUNEL) staining technique was used to examine the apoptosis of retinal ganglion cells in the control group and the puerarin treatment group. Results The number of apoptotic cells in 12, 24 and 72h after reperfusion in the puerarin treatment group was obvious lower than that in the control group(P<0.05). Conclusion Puerarin has protective effects in protecting against apoptosis in ischemia reperfusion injury of rabbit retina.