芪癀胶囊对裸鼠原位移植骨肉瘤的作用及机制研究

发布时间：2018-03-25 01:14

本文选题：芪癀胶囊　切入点：骨肉瘤　出处：《福建中医药大学》2014年硕士论文

【摘要】：目的：观察芪癀胶囊对裸鼠原位移植骨肉瘤生长的作用,并探讨其机制,为扶正祛邪法应用于骨肉瘤治疗提供依据。方法：采用32只4周龄BALB/c-nu/nu裸鼠,麻醉后右侧胫骨注射JMR106-01细胞悬液(5×106个/100μl)制成骨肉瘤原位移植模型,模型鉴定成功后随机分为四组：生理盐水组(1ml生理盐水灌胃,b.i.d)、芪癀胶囊组(1ml芪癀胶囊溶液灌胃,15.6mg/ml, b.i.d)、甲氨蝶呤组(1ml甲氨蝶呤溶液灌胃,3.9mg/ml,q.o.w)、联合治疗组(按芪癀胶囊及甲氨蝶呤各自给药方法灌胃),每组8只,连续灌胃4周。观察四组裸鼠骨肉瘤生长情况：每周测量一次原位骨肉瘤体积,绘制骨肉瘤的生长曲线,计算抑瘤率：其次,4周后取原位骨肉瘤组织,常规切片,HE染色,镜下观察骨肉瘤病理学情况；第三,通过免疫组化二步法检测各组裸鼠原位瘤块中Survivin, VEGF蛋白的表达情况。结果： 1.骨肉瘤生长抑制情况：与生理盐水组相比,其他三组的骨肉瘤生长均有明显的减缓趋势(P0.05),其中,芪癀胶囊加甲氨蝶呤联合抑制骨肉瘤生长疗效最佳；芪癀胶囊组、甲氨蝶呤组和联合治疗组抑瘤率分别是18.01%、24.64%、37.90%,两两比较P0.05。 2.四组原位骨肉瘤Survivin、VEGF蛋白的表达情况：Survivin平均灰度值(IOD)分别为：73.39±1.27、49.29±0.91、37.98±0.85、23.04±0.53；VEGF的IOD值分别为：85.28±1.42、55.04±1.07、34.96±0.89、18.21±0.42。三治疗组与生理盐水组相比IOD值明显降低,P0.05；两两比较P0.05。结论：芪癀胶囊对原位移植骨肉瘤具有抑制作用,芪癀胶囊与甲氨蝶呤联合治疗时,两者具有协同作用,使抑瘤效果更佳。其机制可能是通过抑制骨肉瘤细胞中Survivin和VEGF的高表达,从而诱导骨肉瘤细胞凋亡、降低骨肉瘤血管内皮细胞的增殖和侵袭,从而达到抑制骨肉瘤生长的效果。
[Abstract]:Objective: to observe the effect of Qihuang capsule on the growth of orthotopic transplantation osteosarcoma in nude mice and to explore its mechanism. Methods: the orthotopic transplantation model of osteosarcoma in 32 4-week-old BALB/c-nu/nu nude mice was established by injecting 5 脳 106 / 100 渭 l of JMR106-01 cell suspension into the right tibia after anesthesia. After successful identification of the model, the rats were randomly divided into four groups: saline group (1 ml) was given intragastric infusion of normal saline (1 ml), Qihuang capsule group (1 ml) was gavaged with 15.6 mg / ml, b.i.dg, methotrexate group (1 ml) of methotrexate solution was intragastrically perfused with 3.9 mg 路ml 路ml q.o.wn, and the combined treatment group (Qihuang capsule and Qihuang capsule) was treated by gavage. Methotrexate was administered intragastrically with 8 rats in each group. The volume of osteosarcoma in situ was measured once a week, the growth curve of osteosarcoma was drawn, and the tumor inhibition rate was calculated. The pathological changes of osteosarcoma were observed under microscope. Thirdly, the expression of survivin and VEGF protein was detected by immunohistochemical two-step method. Results:. 1. Growth inhibition of osteosarcoma: compared with normal saline group, the growth of osteosarcoma in the other three groups had a significant slowing trend (P 0.05), among which Qihuang capsule combined with methotrexate had the best effect on inhibiting the growth of osteosarcoma, and Qihuang capsule had the best effect on inhibiting the growth of osteosarcoma. The inhibitory rate of methotrexate group and combined treatment group were 18.01 and 24.647.90, respectively (P 0.05). 2. The expression of survivin protein in osteosarcoma in situ was: the mean gray value of survivin was: 73.39 卤1.270.27, 49.29 卤0.91g, 37.98 卤0.853.04 卤0.53. the IOD value of VEGF in the four groups was: 85.28 卤1.42n 55.04 卤1.0734.96 卤0.8918.21 卤0.422.The IOD value of the three treatment groups was significantly lower than that of the normal saline group (P0.055.05). Conclusion: Qihuang capsule has inhibitory effect on orthotopic transplantation of osteosarcoma, and Qihuang capsule and methotrexate have synergistic effect in the treatment of osteosarcoma. The mechanism may be to inhibit the growth of osteosarcoma by inhibiting the high expression of Survivin and VEGF in osteosarcoma cells, thus inducing the apoptosis of osteosarcoma cells and reducing the proliferation and invasion of vascular endothelial cells in osteosarcoma.
【学位授予单位】：福建中医药大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R273;R285.5

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