脑瘤散对脑胶质瘤细胞GFAP及Bax/Bcl-2的影响

Effect of Naoliu powder on expression of GFAP and BAX/BCL-2 in G422 glioma cells

关键词:  脑瘤散  脑胶质瘤细胞  胶质纤维酸性蛋白  Bax基因  Bcl-2基因

Key words:  NaoLiu powder  glioma cells  glial fibrillary acidic protein  Bax gene  Bcl-2 gene

摘要:

[目的] 观察脑瘤散加环磷酰胺(CTX)对荷瘤鼠体内G₄₂₂脑胶质瘤细胞胶质纤维酸性蛋白(GFAP)表达的影响。[方法] 将G₄₂₂脑胶质瘤细胞株按4 mL:1 g的比例在组织研磨器中研磨成匀浆, 接种于小鼠右前肢腋下皮下, 每只接种0.2 mL.将60只小鼠于接种后次日按体质量分层随机分配法分为3组, 每组20只, 模型组给予脑瘤散加CTX, 阳性对照组给予CTX, 空白对照组只注射生理盐水。给药11 d后处死动物, 取肿瘤标本固定后切片, 免疫组织化学染色测定GFAP的含量, IDA-高清晰度数码显微图象分析系统检测阳性细胞的平均光密度及平均灰度, 同时计数阳性细胞数。免疫组织化学染色测定Bax, Bcl-2表达蛋白含量, 统计其阳性细胞率。[结果] 模型组、阳性对照组的GFAP表达较空白组明显增加, 模型组与阳性对照组之间差异明显, 差异有统计学意义。实验组Bcl-2蛋白的表达明显低于西药组和空白组, 实验组Bax蛋白的表达明显高于西药组和空白组。[结论] 脑瘤散与CTX联合应用, 使肿瘤细胞的损伤加重, 导致细胞坏死凋亡, 其机制可能与上调Bax蛋白的表达, 同时Bcl-2蛋白表达下降, 从而促使G₄₂₂脑胶质瘤细胞凋亡有关。

Abstract:

[Objective] To investigate the effects of Naoliu powder combined with Cyclophosphamide (CTX) on the expression of GFAP and BAX/BCL-2 in glioma cells of tumor-loading mice of G₄₂₂. [Methods] Homogenize of G₄₂₂ glioma cell was made by grinding them in grinding ground in proportion of 4 mL:1 g. They were inoculated into right forelimb of mice subcutaneously. Each mouse was vaccinated with 0.2 mL. Sixty mice were stratified randomly divided into 3 groups (20 mice each group) according to the weight mass at the next day after vaccination. The model group was given with Naoliu powder combined with CTX; positive control group was given with CTX; blank control group was given with normal saline only. Eleven days later the mice were killed and the tumor specimens were harvested, fixed and made into slice. The content of glial fibrillary acidic protein (GFAP) and Bcl-2/BAX was measured with immunohistochemical staining. IDA high definition digital microscopic image analysis system was used to test the mean OD and average gray density in GFAP positive cells. At same time the number of positive cell was detected. [Results] The expression of GFAP in model group and positive control group increased significantly compared with blank group. There was an obvious difference between model group and positive control group with statistical significance. The expression of Bcl-2 protein in the experimental group was significantly lower than Western medicine group and blank group, Bax protein expression in the experimental group was significantly higher than the Western medicine control group and blank group. [Conclusion] Naoliu powder combined with CTX can exacerbate the damage of tumor cells, leading to cell necrosis and apoptosis. The mechanism maybe associated with upregulation of the expression of glioma Bax protein, decreasing the expression of Bcl-2 gene simultaneously and thus contributing to the apoptosis of G₄₂₂ glioma cell.