【摘要】 目的:分析中药薤白挥发油对人胃癌细胞的诱导凋亡作用及其途径,为胃癌患者康复期的中药辅助治疗提供理论依据。方法:实验于2005-04/08在哈尔滨医科大学微生物教研室病毒实验室完成。①胃癌细胞SGC-7901的生长抑制实验:将对数生长期的胃癌细胞以每孔5×104加入培养板中,每孔体积100μL,4个实验孔加入不同浓度穴依次为50,100,200,400mg/L雪的薤白挥发油(由薤白的干燥鳞茎提取),以二甲基亚砜作为阴性对照。采用噻唑蓝比色法检测吸光度,细胞生长抑制率=穴1-实验组吸光度雪/对照组吸光度×100%。应用流式细胞仪检测细胞凋亡率。②免疫细胞化学法检测p53,bcl-2蛋白表达:将对数生长的胃癌细胞SGC-7901以每孔2×105加入培养板,加入薤白挥发油(200mg/L);在对照孔中加入等量的细胞培养液。应用免疫细胞化学法进行细胞凋亡的定性与定量指标测定。结果评估:p53免疫反应定位于细胞核,bcl-2免疫反应定位于胞质和胞膜,细胞染色呈棕褐色或棕黄色为阳性,连续观察5个高倍视野,计算阳性细胞百分率。结果:①薤白挥发油50,100,200,400mg/L组SGC-7901细胞的生长抑制率分别为15.02%,32.33%,60.17%和93.41%;细胞凋亡率分别为10.03%,21.33%,47.65%和88.78%,均显著高于对照组(F=5.93,5.84,P<0.05)。②薤白挥发油组SGC-7901细胞中p53蛋白的阳性率显著高于对照组眼(56.06±7.31)%,(10.23±5.19)%(F=5.63,P<0.05)演。但两组bcl-2蛋白表达差异无显著性意义(P>0.05)。结论:薤白挥发油对人胃癌细胞的杀伤作用主要是通过诱导细胞凋亡来实现,且这种凋亡作用呈剂量依赖性,提升p53蛋白表达可能是诱发胃癌细胞凋亡的作用途径之一。更多还原

【Abstract】 AIM:To analyze the apoptotic effect induced by bulbus allii macrostemi volatile oil on SGC-7901 gastric cancer cells and its approach,and provide theoretic basis for adjunctive therapy in patients with gastric cancer during the rehabilitation period. METHODS: The experiment was performed at the virus laboratory,Department of Microorganism,Harbin Medical University from April to August 2005. ①Growth inhibiting trial of SGC-7901 gastric cancer cells: The gastric cancer cells in logarithmic phase was kept in culture plate,100 μL 5×104 cells in each hole. The bulbus allii macrostemi volatile oil (extracted from dried squamous bulb of bulbus allii macrostemi) of different concentration (50,100,200 and 400 mg/L in order) was put into the 4 experimental holes. Dimethyl sulfoxide (DMSO) was used as negative control. Absorbance (A) was detected with MTT. Cell growth inhibiting rate referred to (1-A of experimental group)/A of control group×100%. Apoptosis rate was determined with flow cytometer. ②The p53 and bcl-2 protein expression detected with immunocytochemical method: The SGC-7901 gastric cancer cells in logarithmic phase was kept in culture plate,2×105 cells in each hole,and added with 200 mg/L bulbus allii macrostemi volatile oil. Cell culture fluid of the same volume was added in the control hole. Qualitation and quantitation of apoptosis were determined with immunocytochemical method. Result evaluation: immune reaction of p53 located at nucleus,while immune reaction of bcl-2 located at kytoplasm and cell membrane. Brown or buffy showed the cell was positive. After observing continuously for 5 high power sights,the percentage of positive cells was calculated. RESULTS: ①Growth inhibiting rates of SGC-7901 cell in the 50,100,200,400 mg /L bulbus allii macrostemi volatile oil groups were 15.02%,32.33%,60.17% and 93.41%,respectively. Cell apoptosis rates were 10.03%,21.33%,47.65% and 88.78%,respectively. They were significantly higher than those of the control group (F=5.93,5.84,P < 0.05). ②Positive rate of p53 protein of SGC-7901 cell in the bulbus allii macrostemi volatile oil group was obviously higher than that in the control group [(56.06±7.31)%,(10.23±5.19)%],(F= 5.63,P < 0.05)]. But there was insignificant difference in bcl-2 protein expression between the two groups (P > 0.05). CONCLUSION: The bulbus allii macrostemi volatile oil exerts its cytotoxic effect on SGC-7901 gastric cancer cells by inducing apoptosis. This apoptosis shows dose-dependent effect. Increasing the expression of p53 protein may be one of its mechanisms of inducing gastric cancer cell apoptosis.更多还原