【作者】 冯军；

【导师】 赵洪洋；

【作者基本信息】 华中科技大学 ， 外科学， 2007， 博士

【摘要】 第一部分立体定向建立大鼠C6脑胶质瘤模型目的:应用不同数量C6胶质瘤细胞接种大鼠脑内建立大鼠脑胶质瘤模型,观察其生长特性,比较各种接种量的特点。方法:分别取1.0×10~5个/10μl,1.0×10~6个/10μl,1.0×10~7个/10μl体外培养的大鼠C6胶质瘤细胞单细胞悬液10μl,立体定向接种于Wistar大鼠脑右侧尾状核区,观察不同接种量实验鼠的生存状态、成瘤情况及脏器转移灶,并采用免疫组织化学方法检测其胶质纤维酸性蛋白、血管内皮生长因子的表达和微血管密度。结果:各种接种量的实验组成瘤率均为100%,未见颅外转移病灶,瘤细胞病理性核分裂像多见,胶质纤维酸性蛋白呈散在阳性表达,血管内皮生长因子呈强阳性表达,瘤组织内有出血和坏死以及丰富的微血管,在组织病理学上接近人脑胶质瘤。结论:C6细胞立体定向接种Wistar大鼠脑右侧尾状核建立脑深部胶质瘤动物模型,大鼠成瘤率高,颅内生长稳定,肿瘤组织病理学及形态学特性与人脑胶质瘤相似,可作为临床胶质瘤基础研究的理想模型。实验范围内部分时段各种接种剂量肿瘤生长速度差异有显著性意义,可根据病因学、实验治疗或药效学等不同研究需要选择不同接种量。第二部分C6胶质瘤大鼠放射性脑水肿与CD18mRNA表达的关系目的:研究C6胶质瘤大鼠放射性脑损伤后脑水肿的程度以及与血液中性粒细胞CD18mRNA表达水平改变的关系。方法:雄性Wistar大鼠40只,体质量250 g -300 g,随机分成A、B两组,每组20只,A组为接受放射线照射组,B组为对照组。两组均颅内种植胶质瘤C6细胞,A组在种植肿瘤细胞15天后予Co60照射,照射后第7天所有实验大鼠均取全脑测含水量;取1mm3大小正常脑组织和肿瘤组织,电镜下观察毛细血管;颈总动脉取血,离心,取中性粒细胞层,逆转录-聚合酶链式反应分析测定血液中性粒细胞CD18mRNA表达水平。结果:脑组织含水量:A、B两组间差异有显著性意义(P<0.05);血液中性粒细胞CD18mRNA表达水平:A、B两组间差异有显著性意义(P<0.05)。未接受放射线的胶质瘤组织,电镜下见毛细血管管腔比正常脑组织多见。接受放射线的胶质瘤组织毛细血管受到不同程度的损伤,血管周围水肿。结论:放射线可以导致脑水肿的发生,放射性脑水肿的发生与血液中性粒细胞CD18mRNA的改变相关。第三部分甲基强的松龙及地塞米松治疗放射性脑水肿与CD18mRNA表达的关系目的:研究甲基强的松龙与地塞米松对C6胶质瘤大鼠放射性脑损伤后脑水肿程度的影响以及对血液中性粒细胞CD18mRNA表达水平的改变。方法:雄性Wistar大鼠100只,体质量250 g -300 g,随机分成5组,每组20只,分别为接受照射同时30mg/kg/d甲基强的松龙治疗组(A组)、接受照射同时10mg/kg/d甲基强的松龙治疗组(B组)、接受照射同时5mg/kg/d地塞米松治疗组(C组)、接受照射但未接受激素治疗的实验对照组(D组)和未接受照射和激素治疗的正常对照组(E组)。各组均颅内种植肿瘤,A、B、C、D四组于种植15天后给予Co60照射,同时所有实验鼠自照射前3天开始静脉注射激素或生理盐水,直至照射后6d,每天1次,照射后第7天取大鼠脑组织测含水量,颈总动脉取血,离心取中性粒细胞层,逆转录-聚合酶链式反应分析测定血液中性粒细胞CD18mRNA表达水平。结果:脑组织含水量: D组、E组分别与A、B、C组比较差异均有显著性意义(P<0.05); D、E组两组间比较,差异有显著性意义(P<0.05);C组分别与A、B组比较,差异均有显著性意义(P<0.05);A、B两组间比较,差异无显著性意义(P>0.05)。血液中性粒细胞CD18mRNA表达水平:D、E组分别与A、B、C组比较,差异均有显著性意义(P<0.05);D、E组两组间比较,差异有显著性意义(P<0.05);C组与A、B组比较,差异均有显著性意义(P<0.05);A、B两组间比较,差异无显著性意义(P>0.05)。结论:甲基强的松龙与地塞米松均可通过抑制CD18mRNA表达的方式减轻放射性脑水肿,但甲基强的松龙效果较好。第四部分放射性脑水肿的发生与甲基强的松龙治疗的疗效观察目的:研究放射性脑水肿的发病规律和甲基强的松龙与地塞米松治疗放射性脑水肿的效果。方法:回顾63例放射性脑水肿患者,统计各种性质占位所占的比例,统计放射线治疗到出现放射性脑水肿症状的时间,比较甲基强的松龙(A组)和地塞米松(B组)治疗放射性脑水肿前后临床症状改变情况和影像学检查脑水肿范围改变情况。结果:63例患者中,共有脑膜瘤17例(26.98%),动静脉畸形12例(19.05%),海绵状血管瘤12例(19.05%),脑实质内胶质瘤10例(15.87%),脑室内占位6例(9.52%),垂体腺瘤2例(3.17%),听神经瘤2例(3.17%),松果体区占位1例(1.59%),转移瘤1例(1.59%)。放射性脑水肿症状出现于接受放射线治疗后0月-36月,平均8.1个月,A组治疗有效率92.30%,B组治疗有效率88.89%,其差别没有统计学意义(P>0.05),A组治疗前后脑水肿范围差异有显著性意义(P<0.05),B组治疗前后脑水肿范围差异无显著性意义(P>0.05)。结论:窦旁脑膜瘤、海绵状血管瘤、动静脉畸形和胶质瘤等发生放射性脑水肿的危险性较大;放射性脑水肿多发生于患者接受放射线治疗后1年内,尤其是第1个月内和第5个月到第7个月;甲基强的松龙和地塞米松对放射性脑水肿都具有治疗作用,在影像学上,甲基强的松龙效果更好。更多还原

【Abstract】 PartⅠEstablishment of Rat Cell Line C6 Brain Glioma Model with StereotaxisObjectiveTo establish an brain glioma models of C6 cells in Wistar rats with different methods and to study and compare the growth patterns of the tumors formed in the wistar rat brain.MethodsRat C6 glioma cells were cultured in vitro. 10μl of cell suspension containing 1.0×10⁵ C6 cells/10μl or 1.0×10⁶ C6 cells/10μl or 1.0×10⁷ C6 cells/10μl was stereotaxically inoculated into the right caudate nucleus of the rat brain. The moving and survival satates of rats with gliomas were observed. The examinations of the tumors formation, volumes, metastassis and histopathology were performend. The expression of glial fibrillary acidic protein and vascular endothelial growth factor in the tumors and the microvessel density were detected by immunohistochemistry.ResultsAll the tested rats of different inoculate dose developed brain tumors without extracranial metastasis. The rat C6 gliomas resembled histopathclogical features of human glioma. The tumors contained many cells with remarkable cytoplasmic and nuclear polymorphism, and pathological mitosis figures. The scattered positive expression of vascular endothelial growth factor was observed in tumor cells. There was the strongly positive expression of glial fibrillary acidic protein in the tumors.ConclusionsThe successful rate of C6 brain tumor model established by the intracerebral inoculation of C6 cells is very high in rats. The tumor established in the experiment is similar to human glioma in histopathological features and it can be used as a good model to study the glioma. The tumors model established by inoculation of different dose of C6 cells can be selected according to different needs of different experiments because their growth speeds are different from each other. PartⅡThe Relationship Between the Radioactive Cerebral Edema of the Rat of C6 Brain Glioma Model and the Expression of CD18 mRNAObjectiveTo study the extent of radioactive cerebral edema and the expression of CD18 of the rat neutrophilic granulocyte.Methods40 male Wistar rats were divided into 2 groups at random （n=20）. Group A was irradiated with Co60, group B was normal control. Planted glioma cells into brains of rats of group A and B, and treated groups A with Co60 after 15 days. The water content of the brains of the rats and the expression of CD18 of the rats neutrophils were measured 7d after irradiation. Collect the normal brain tissue and the tumor tissue about 1mm3 and observed blood capillary on electron microscope.ResultsThe water content of the rats brain: There were significant differences between group A and B （P<0.05）.The level of CD18 mRNA: There were significant differences between group A and B （P<0.05）. The blood capillary of tumor tissue was much than the normal brain tissue, the blood capillary of tumor tissue of irradiated rat was injuried and have edemata around it.ConclusionRadioactive ray can lead to the brain edema and the development of radioactive cerebral edema is related by change of the expression of CD18 in rats. PartⅢThe Relationship Between the Therapy of Radioactive Cerebral Edema with Methylprednisolone and Dexamethasone and the Expression of CD18 mRNAObjectiveTo study the effect of methylprednisolone and dexamethasone to radioactive cerebral edema and the expression of CD18 of the rat neutrophilic granulocyte. MethodsOne hundred male Wistar rats were divided into 5 groups atrandom （n=20）. Group A was irradiated with 30 mg/kg methylprednisolone, group B was irradiated with 10 mg/kg methylprednisolone, group C was irradiated with 5 mg/kg dexamethasone, group D was irradiated without glucocorticoid, and group E was normal control. Planted glioma cells into brains of rats of group A、B、C、D and E, and treated groups A、B、C、D with Co60 after 15 days. All groups gave glucocorticoid or normal saline beginning 3d before and ending 6d after irradiation（iv, qd）, The water content of the brains of the rats and the expression of CD18 of the rats neutrophils were measured 7d after irradiation.ResultsThe water content of the rats brain: There were significant differences between group D、E and group A、B、C（P<0.05） , group D and group E（P<0.05）, group C and group A、B（P<0.05）; There was no significant difference between group A and B（P>0.05）. The level of CD18 mRNA: There were significant differences between group D、E and groups A、B、C（P<0.05）; group D and group E（P<0.05）, group C and group A、B（P<0.05）; There was no significant difference between group A and B（P>0.05）.ConclusionMethylprednisolone and dexamethasone can both protect the radiation-induced injury and brain edema changes by inhibiting the expression of CD18 in rats. and the methylprednisolone is more efficient than dexamethasone. PartⅣThe Development of Radioactive Cerebral Edema and the Curative Effect of MethylprednisoloneObjectiveTo study the method of radioactive cerebral edema and the effectiveness of methylprednisolone and dexamethasone.Methods63 patients of radioactive cerebral edema were reviewed, the percentage of various kinds entity, the time from radiotherapied to find radioactive cerebral edema were statisticsed. The change of clinical symptom and the ambit of cerebral edema treated with methylprednisolone（group A） or dexamethasone（group B） were compared.ResultsIn 63 patients, have 17 meningeoma（26.98%）, 12 arteriovenous malformation（19.05%）, 12 cavernous angioma（19.05%）, 10 glioma（15.87%）, 6 tumor in cerebral ventricle（9.52%）, 2 hypophyseoma（3.17%）, 2 acoustic nerve tumor（3.17%）, 1 tumor of apophysis cerebri（1.59%）, 1 metastatic tumor（1.59%）. 0mon-36mon from radiotherapied, the symptoms of radioactive cerebral edema appeared. Average is 8.1 month. The effective power of group A is 92.30%, The effective power of group B is 88.89%, showing no significant difference. But other than in group B, significant differences of ambit of cerebral edema between pretherapy and post-treatment were detected in group A（P<0.05）.ConclusionsParasagittal sinus meningioms, arteriovenous malformation, cavernous angioma, glioma and tumor in cerebral ventricle have more chance of radioactive cerebral edema. Radioactive cerebral edema often betide 1 year after radiotherapiy, especially in 1st month and 5^th mon-7^th mon. Methylprednisolone and dexamethasone can both protect the radiation-induced injury. And the methylprednisolone is more efficient than dexamethasone in imageology.更多还原