摘要
目的探讨硫化氢（H：S）在大鼠恶性胶质瘤生长中的作用及其机制。方法将40只雄性成年sD大鼠随机分为对照组、硫氢化钠（NariS）组、肿瘤组、肿瘤＋NaHS组，每组10只。对照组脑内注射人工脑脊液1周后，腹腔注射生理盐水；NaHS组脑内注射人工脑脊液1周后，腹腔注射NaHS溶液；肿瘤组脑内注射c6细胞悬液1周后，腹腔注射生理盐水；肿瘤＋NaHS组脑内注射c6细胞1周后，腹腔注射NaHS。每组大鼠均正常饲养3周，观察记录大鼠的一般状况和体重增长情况。处死大鼠，测定大鼠脑组织中H：S的含量，组织病理学检查瘤体细胞形态变化并测量瘤体体积，免疫组化法检测缺氧诱导因子lot（HIF．1ct）和CD34的表达以及瘤内微血管密度（MVD）。结果与对照组相比，NariS组各项指标无显著变化；肿瘤组大鼠一般状况明显较差，体重增加缓慢；肿瘤＋NaHS组大鼠机体消耗症状明显加重，部分大鼠体重出现负增长。肿瘤组大鼠瘤体体积为（32．0±6．9）mm。，肿瘤＋NaHS组为（67．84-11．9）mm3，NaHS处理后的大鼠瘤体体积显著增大（P〈0．001）。与肿瘤组相比，肿瘤＋NariS组大鼠肿瘤组织中HIF-let和CD34表达量均显著增加。肿瘤组和肿瘤＋NaHS组大鼠肿瘤组织中MVD分别为（41．2±7．9）／mm。和（97．04-10．8）／mm2，差异有统计学意义（P〈0．001）。对照组、NaHS组、肿瘤组、肿瘤＋NaHS组大鼠脑内H：S含量分别为（29．12±0．94）nmol／g、（29．73±O．76）nmol／g、（35．25±1．03）nmol／g和（40．81±1．21）nmol／g。统计分析显示，对照组和NaHS组H＇s含量差异无统计学意义（P〉0．05），肿瘤组H：S含量显著高于对照组（P〈0．05），肿瘤＋NailS组含量显著高于肿瘤组（P〈O．05）。结论恶性胶质瘤的生长伴随内源性H：S含量的增加，外源性H，S能促进恶性胶质瘤的生长，其机制与H：S促进肿瘤细胞增殖和瘤内新生血管形成有关。
Objective To address the hypothesis that hydrogen sulfide （H2S） is a functionally significant stimulator in the development of glioblastoma （GBM） and explore the mechanism of stimulation. Methods Forty adult Sprague-Dawley （SD） rats were given intracerebral injection of rat C6 glioma cell suspension, and an intraperitoneal injection of sodium hydrosulfide （NariS） , an exogenous H2S donor. The 40 rats were randomly divided into 4 groups of 10 rats in each： the control group, NariS group, C6 glioma group （intracerebral implantation of C6 glioma cells） and C6-NaHS group （intracerebral implantation of C6 glioma cells and intraperitoneal injection of NariS）. Food and water were freely available during all phases of the experiment. Physical symptoms were observed and the tumor size was measured. Histological changes were examined by pathology. Immunohistochemical staining was used to analyze the expression of HIF-lot and integrated optical density （IOD） was used to determine the tumor microvessel density （MVD）. The H2S content in the brain was measured. Results The physical symptoms of tumor-bearing rats became more serious after NariS injection. The H2S level in the C6 glioma group was higher than that in the control group [ （35.25 ±1.03 ） nmol/g vs. （ 29.12 ± 0.94） nmol/g, P 〈 0.05 ], and the highest H2 S level was found in the C6-NaHS group. The pathological examination showed that the implanted tumors were predominantly spheroid with a distinct border and no capsule could be detected. Neovascular proliferation was also observed. Foci of tumor necrosis, intratumoral hemorrhage, pseudopalisades and tumor cavity were clearlyobserved. The glioma cells had scant eosinophilic cytoplasm and enlarged hyperchromatic nuclei. All these phenomena were more markedly in the C6-NaHS group compared with that in other three groups. The mean tumor volume was significantly different between the C6 and C6-NaHS rats [ （32. 0 ± 6. 9） mm3 vs. （67.8 ± 11.9） mm3, P 〈 0. 001 ]. Immunohistoehemical analysis exhibited that the hypoxia-inducible factor-lalpha （HIF-la） and CD34 expression were significantly increased after the intraperitoneal injection of NariS in the C6-NaHS rats （comparing the IOD between C6-NaHS group and C6 group, H1F-la： 133 962.9 ± 451.4 vs. 38 569.8 ± 408.6, P 〈 0. 001 ; CD34 ： 73 368.6 ±404.8 vs. 14 570.6 ± 748.7, P 〈 0. 001 ）. Moreover, compared with the C6 group, there were higher MVD in the C6-NaHS group [ （41.2 ± 7.9）/ramz vs. （97.0 ±10.8）/ram2, P 〈 0.001 ]. Conclusions H2 S serves as a stimulator in the development of rat glioblastoma and exogenous Hz S strongly promotes the tumor growth. The stimulating mechanisms include the increase of HIF-lot expression and neovascular formation. H2S may be a significant regulator in the development of
出处
《中华肿瘤杂志》
CAS
CSCD
北大核心
2012年第4期254-258,共5页
Chinese Journal of Oncology
基金
国家自然科学基金（31171053）
