【作者】 蒲娅；

【导师】 高永翔；

【作者基本信息】 成都中医药大学 ， 中西医结合基础， 2009， 硕士

【摘要】 目的：研究青藤碱干预肿瘤细胞凋亡的信号转导机制。观测不同剂量青藤碱干预肿瘤细胞凋亡的效果。为青藤碱在肿瘤的治疗提供实验依据。方法：实验动物随机分高、中、低3个剂量组及空白对照组,传代接种后7d的S180瘤源小鼠,断椎处死,无菌条件下抽取瘤液,放入无菌容器内,加1：3无菌生理盐水,每只小鼠均右腋皮下接种肿瘤细胞悬液0.2 mL整个操作在30 min内完成。肿瘤种植24 h后随机分成青藤碱高、中、低剂量组,对照组注射生理盐水,每组10只,每天注射1次,连续给药7d。每日观察小鼠生长情况。停药次日断椎处死,称取体重及瘤重。用公式计算抑瘤率。取少量瘤组织制备单细胞悬液,用western blot方法检测青藤碱不同剂量对COX-2、ERK、BCL-2蛋白的表达情况。用流式细胞术检测S180腹水瘤细胞的凋亡。结果：1.空白对照组的平均瘤重为1.415±0.30g,中高剂量组平均瘤重分别为0.799±0.34g、0.625±0.16g,中高剂量组抑瘤率分别为43.4%(p<0.05)、55.8(p<0.01)。说明大剂量的青藤碱明显抑制了小鼠移植S180瘤细胞的生长。2.中高剂量的青藤碱可抑制COX-2蛋白表达。与空白组比较,实验组COX-2蛋白表达下降(P<0.05)；3.中高剂量实验组p-ERK蛋白表达下降。与空白组比较,(P<0.05)。低剂量组下降不明显；4.中高剂量实验组BCL-2蛋白表达下降明显。与空白组比较,(P<0.05)；5.与空白组比较,实验组细胞凋亡增加(P<0.05),说明青藤碱作用后,细胞凋亡率升高。结论：1.选择性COX-2抑制剂青藤碱对S180瘤细胞的增殖具有抑制作用；2.青藤碱能够抑制P-ERK的表达,诱导S180瘤细胞凋亡；3.低毒性且容易从中药中提取的选择性COX-2抑制剂青藤碱有望在肿瘤的治疗发挥重要作用,具有良好的应用前景。更多还原

【Abstract】 Goal:The research cocculus tunbergii alkali intervention tumour cell perishes weakly signal transduction mechanism. Observes effect which the different dosage cocculus tunbergii alkali intervention tumour cell perishes weakly. Provides the experiment for the cocculus tunbergii alkali in the tumor prevention and the treatment to rest on.method:Experimental animal along with fuselage high, middle and low 3 monitoring teams and a blank control group, after transfer of generation vaccination, the 7d S180 lump source mouse, breaks zhui the execution, under the aseptic condition extracts the lump fluid, puts in the aseptic vessel, adds 1:3 aseptic physiological saline, each mouse right armpit hypodermic vaccination tumour cell hangs fluid 0.2 mL, the entire operation completes in 30 min. After the tumor plants 24 h, divides into the cocculus tunbergii alkali high, middle and low monitoring team stochastically, the control group, the control group injects the physiological saline, each group of 10, inject every day 1 time, gives medicine 7 d continuously. Observes the mouse growth situation every day. Stops the medicine to break zhui the execution next day, the name takes the body weight and the lump is heavy. Damps the lump rate with the formula computation. Takes the few lump tissue preparation single cell to hang the fluid, method examines the cocculus tunbergii alkali different dosage with western the blot to COX-2, AKT, the Caspase3 protein expression situation. Examines the S180 ascites lump cell with the class type cell technique to perish weakly.Conclusion:1. the selective COX-2 inhibitor cocculus tunbergii alkali has the inhibitory action to the S180 lump cell’s multiplication; 2. the cocculus tunbergii alkali can suppress P-ERK the expression, responded through the Caspase family cascade, activates Caspase-3 the activeness, induces the S180 lump cell to perish weakly; 3. and the low toxicity withdraws easily from the traditional Chinese medicine the selective COX-2 inhibitor cocculus tunbergii alkali hopefully plays the influential role in the tumor prevention and the treatment, has the good development application prospect.更多还原