论文部分内容阅读
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 of exogenous hydrogen sulfide (H2S) on glioma angiogenesis in rats. Methods Forty adult SD rats were randomly divided into 4 groups: control group, sodium hydrosulfide (Na HS) group, tumor group and tumor + Na HS group, with 10 rats in each group. Tumor group, tumor + Na HS group, C6 glioma cells were injected into the striatum of rats to establish glioma model; control group, Na HS group, rat brain without any treatment. The rats in the four groups were sacrificed for 3 weeks and then decapitated. The contents of H2S in the brain tissue and tumor in each group were detected by the sensitive sulfur electrode method. The levels of cystathionine-β-synthase (CBS) and 3-mercapto-pyruvate sulfur transferase (3-MST) were detected by ELISA. Western blot and ELISA were used to detect the expression of HIF-1α, VEGF, Metalloproteinase-9 (MMP-9) expression; while counting intratumoral microvessel density (CD105-MVD). Results In the tumor group and NaHS group, obvious tumor tissues were observed. Compared with the control group, the number of capillary vessels in the tumor group was significantly increased. In the tumor + Na HS group, there were more HIF-α, VEGF, MMP -9 expression, and MVD was significantly higher than the control group, Na HS group, tumor group (all P <0.01). Conclusions Exogenous H2S can induce angiogenesis in rat glioblastoma. The possible mechanism is that H2S may promote the expression of CBS, 3-MST, HIF-α, VEGF and MMP-9.