苦参碱联合替莫唑胺对大鼠脑胶质瘤MMP2、MMP9、VEGF表达的影响

目的探讨苦参碱联合替莫唑胺对大鼠脑胶质瘤基质金属蛋白酶(MMP)-2、MMP-9、血管内皮生长因子(VEGF)表达的影响。方法建立大鼠C6脑胶质瘤模型,采用MRI扫描、HE染色、免疫组织化学确定模型建立成功。将C6脑胶质瘤模型大鼠随机分成4组,每组各20只:(1)对照组,腹腔注射生理盐水(100 ml/kg);(2)苦参碱组,腹腔注射苦参碱(20 mg/kg);(3)替莫唑胺组,腹腔注射替莫唑胺(25 mg/kg);(4)联合组,腹腔注射苦参碱(20 mg/kg)和替莫唑胺(25 mg/kg);每天给药1次,连续14 d。药物治疗后14 d处死大鼠,取胶质瘤组织,采用免疫印迹法和实时荧光定量PCR检测MMP-2、MMP-9和VEGF蛋白和m RNA表达情况。结果大鼠注射胶质瘤C6细胞后7 d,MRI扫描可见脑胶质瘤的存在,HE染色发现组织表现出神经胶质瘤的侵袭性生长和细胞核分解,免疫组织化学结果显示也证明大鼠存在脑胶质瘤。与对照组相比,苦参碱组、替莫唑胺组与联合组MMP-2、MMP-9、VEGF蛋白和m RNA表达水平均显著降低(P0.05);联合组MMP-2、MMP-9、VEGF蛋白和m RNA表达水平均显著低于苦参碱组和替莫唑胺组(P0.05),而苦参碱组MMP-2、MMP-9、VEGF蛋白和m RNA表达水平均显著低于替莫唑胺组(P0.05)。结论苦参碱和替莫唑胺均可显著降低大鼠脑胶质瘤MMP-2、MMP-9、VEGF表达水平,而且,二者联用作用更强。

Effects of matrine combined with temozolomide on the expression of MMP2, MMP9 and VEGF in glioma tissues in rats with gliomas

Objective To investigate the effects of matrine combined with temozolomide on the expressions of matrix metalloproteinases (MMP)2, MMP9 and vascular endothelial growth factor (VEGF) in glioma tissues in rats. Methods C6 glioma models were established in 80 rats. The glioma models were evaluated by MRI, HE staining and immunohistochemical technique. The rats with C6 gliomas were randomly divided into 4 groups of 20 animals each, i.e. control group in which the rats were treated only by physiological saline, matrine group in which the rats were treated only by matrine, temozolomide group in which the rats were treated only by temozolomide, and combination group in which the rats were treated by matrine and temozolomide. The solution of drug corresponding to each group was intraperitoneally injected once a day for 14 days. The expressions of MMP2, MMP9 and VEGF protein and mRNA in the glioma tissues were detected respectively by western blot and qRT-PCR in all the groups. Results The successful establishment of C6 cell glioma model was proved by MRI, HE staining and immunohistochemical technique. The levels of MMP2, MMP9, VEGF protein and mRNA expressions were significantly lower in the combination group than those in matrine group (P<0.05), which were significantly lower than those in temozolomide group (P<0.05), which significantly lower than those in the control group (P<0.05). Conclusions Matrine can enhance the chemotherapeutic effect of temozolemide on gliomas in the rats and matrine combined with temozolemide can significantly reduce the expressions of MMP2, MMP9, VEGF protein and mRNA in the glioma tissues in the rats.