外源性硫化氢诱导大鼠神经胶质瘤血管形成机制的研究

Study mechanism induced by exogenous hydrogen sulfide on promotes and stimulates the angiogenesis of rat glioblastoma

目的研究外源性硫化氢(H2S)对大鼠胶质瘤血管形成的诱导机制。方法成年SD大鼠40只,随机分为4组,对照组、硫氢化钠(Na HS)组、肿瘤组、肿瘤+Na HS组,每组10只。肿瘤组、肿瘤+Na HS组,大鼠纹状体内注射C6胶质瘤细胞,建立胶质瘤模型;对照组、Na HS组,大鼠大脑不做任何处理。4组大鼠正常饲养3周后,对其进行断头取脑,用敏感硫电极法检测各组大鼠脑组织及瘤内H2S的含量;Real time PCR法检测胱硫醚-β-合成酶(CBS)、3-巯基丙酮酸硫转移酶(3-MST)的mRNA转录水平;ELISA及蛋白免疫印迹法检测缺氧诱导因子-1α(HIF-1α)、血管内皮生长因子(VEGF)、基质金属蛋白酶-9(MMP-9)的表达;同时计数瘤内微血管密度(CD105-MVD)。结果肿瘤组、肿瘤+Na HS组,均可见明显的肿瘤组织,与对照组相比,瘤内毛细血管数明显增多;肿瘤+Na HS组,瘤体内有更多的HIF-α、VEGF、MMP-9的表达,且MVD明显高于对照组、Na HS组、肿瘤组(均P<0.01)。结论外源性H2S能诱导并促进大鼠恶性胶质瘤内的血管生成,其发生机制可能与H2S促进CBS、3-MST、HIF-α、VEGF及MMP-9的表达密切相关。

Objective To study the mechanism induced by exogenous H2 S on rat glioma formation. Methods Fourty adult SD rats were ran- domly divided into control group, Naris group, tumor group, tumor ＋ Naris group of four groups, each group 10. Tumor group, tumor ＋ Naris group injected rat striatum C6 glioma cells to established glioma model. Control group, Naris group received no treatment. Four groups of rats fed normally, three weeks after taking the brain sensitive sulfur electrode applied to detect the rat brain tumor and HzS content of its decapitated. The cystathionine - beta - synthase （ CBS）, mRNA transcription level of 3- mercaptopyruvate sulfurtransferase （3MST） were determined by real time PCR assay. The hypoxia - inducible transcription factors - 1α （ HIF - 1α）, vascular endothelial growth factor （VEGF） and the expre- ssion of matrix metallo proteinase - 9 （ MMP - 9 ） by ELISA and Western blot. The intratumoral micro vessel density （MVD） was counted by CD105- MVD. Results Compared with the control group, Naris group, tumor group, tumor ＋ Naris group were clearly visible tumor tissue, significantly increased the number of tumor capillaries. Tumor ＋ Naris group has more CBS, 3MST, HIF - α, VEGF expression of MMP - 9. And tumor ＋ Naris group was significantly higher than the Naris group, tumor group （P 〈 0.01 ）. Conclusion Exogenous H2S may induce and promote angiogenesis in rat glioma within its mechanism of H2S promote CBS, 3MST, closely related to the HIF-α VEGF expression of matrix metalloproteinase MMP- 9.