CD39在普萘洛尔治疗血管瘤裸鼠模型中的作用机制探讨

发布时间：2018-06-30 02:16

本文选题：CD39 + 血管瘤　；参考：《泸州医学院》2013年硕士论文

【摘要】：目的：建立婴幼儿增殖期血管瘤的裸鼠模型，探讨CD39在普萘洛尔治疗增殖期血管瘤中的作用机制。方法：将在手术室无菌条件下手术切除的婴幼儿增生期毛细血管瘤新鲜标本，均匀切成小块（约3.0mm×3.0mm×3.0mm）移植入实验裸鼠臀背部皮下，每只皮下移植两个瘤体组织，建立婴幼儿血管瘤的裸鼠异种移植的动物实验模型。在移植45天后，将移植组织均成活的60只的实验荷瘤裸鼠进行随机对照分组实验，随机分为两个实验小组：组A（普萘洛尔治疗组），组B（生理盐水对照组），每组30只。无菌条件下，分别给予普萘洛尔溶液和生理盐水灌胃进行干预，隔日一次。在首次给予药物治疗前、治疗后7天、14天、21天、28天，使用颈椎脱臼法，分批将实验裸鼠处死，每次每组随机处死6只，切取瘤体组织。用游标卡尺分别测量两组实验裸鼠移植瘤横径b的和最大直径a,并根据公式V=π／6×a×b2估算肿瘤体积变化情况；将切取的瘤体组织石蜡包埋，制成切片，进行HE染色，分析瘤体组织特点，同时进行免疫组织化学染色，检测组织的CD39、Caspase-3和Ki-67的表达；TUNEL法检测移植瘤组织中的细胞凋亡情况；采用Pearson相关分析法进行CD39表达光密度值与细胞凋亡的相关性分析；酶联免疫吸附法（ELISA）检测移植瘤组织的ATP浓度；所有的实验数据统计学分析使用SPSS19.0统计软件进行。结果：治疗前，各组的实验裸鼠的血管瘤组织都生长良好，组间瘤体的体积无显著差异（P＞0.05）。治疗后7天、14天，生理盐水对照组的血管瘤瘤体体积继续生长、增大，而普萘洛尔治疗组血管瘤瘤体体积明显小于生理盐水对照组（P0.01）。干预后21天、28天，普萘洛尔治疗组瘤体进一步缩小，较前更加明显，而生理盐水组裸鼠瘤体也开始不同程度的缩小，但是幅度较小。在镜下观测瘤体标本HE染色切片，普萘洛尔治疗组瘤体组织血管减少、破坏，管间纤维脂肪组织浸润，血管内皮细胞分散、稀疏、萎缩，血管壁塌陷、变形，，管腔有闭塞、破坏征象；而生理盐水对照组血管瘤瘤体组织毛细血管丰富，密集，未见管腔和血管壁有萎缩、破坏征象。普萘洛尔治疗组的CD39光密度（MD）和Ki-67的阳性指数与生理盐水对照组相比均有减少，两组间有显著差异性（P0.05）。普萘洛尔治疗组的Caspase3和细胞凋亡指数明显高于生理盐水组，两组间有显著差异性（P0.01）；ELISA法检测ATP浓度，显示普萘洛尔治疗组高于生理盐水对照组，两组之间有显著差异性（P0.05）。 CD39表达与瘤体细胞凋亡呈负相关（r=-0.649,P0.01）。结论：1.普萘洛尔可促进婴幼儿增生期血管瘤裸鼠模型中的瘤体消退。2.普萘洛尔对增生期血管瘤的治疗机制与血管瘤内皮细胞上CD39的表达变化有关。可能其通过降低CD39对ATP的降解能力，发挥ATP的细胞毒性作用，从而促使瘤体细胞凋亡，达到治疗作用。
[Abstract]:Objective: to establish a nude mouse model of infantile proliferative hemangioma and explore the mechanism of CD39 in propranolol in the treatment of proliferative hemangioma. Methods: a fresh specimen of neonatal hyperplastic capillary hemangioma was removed under aseptic conditions in the operation room and cut into a small block (about 3.0mm x 3.0mm x 3.0mm) and transplanted into the hip back of nude mice Subcutaneous, each subcutaneous transplantation of two tumor tissues to establish an experimental model of xenotransplantation in nude mice of infantile hemangioma. After 45 days of transplantation, 60 experimental tumor bearing nude mice were randomly divided into two experimental groups: group A (propranolol treatment group) and group B (saline control) Group 30. Under aseptic conditions, propranolol and saline were given to the stomach for intervention, once every other day. Before the first drug treatment, 7 days, 14 days, 21 days, 28 days after the treatment, the experimental nude mice were executed in batches, and 6 rats were killed each time, and the tissues were measured with vernier calipers. The two groups tested the transverse diameter of B and the largest diameter of a in nude mice. According to the formula V= pi / 6 x a x B2, the tumor volume changes were estimated. The cut tumor tissues were embedded in paraffin, made into slices, stained with HE, and analyzed the tissue characteristics of the tumor. At the same time, immunohistochemical staining was carried out to detect the expression of CD39, Caspase-3 and Ki-67 in the tissue; TUNEL The apoptosis of the transplanted tumor tissue was detected by the method of Pearson correlation analysis. The correlation analysis between the CD39 expression of light density and the apoptosis was analyzed; the ATP concentration of the transplanted tumor tissue was detected by enzyme linked immunosorbent assay (ELISA); all the experimental data were statistically analyzed using SPSS19.0 software. Results: before treatment, each The hemangioma tissue of the experimental nude mice grew well, and there was no significant difference in the volume between the groups (P > 0.05). The volume of hemangioma in the saline control group continued to grow and increase on the 7 day after treatment, and the volume of the hemangioma in the propranolol group was significantly smaller than that of the saline control group (P0.01). The prognosis of the propranolol group was 21 days, 28 days, and proprannaphthalene after the treatment. The tumor in the group of lol treatment was further narrowed and more obvious than before, but the tumor body of nude mice in the saline group began to shrink in varying degrees, but the range was smaller. The HE staining section of the tumor specimen under the microscope showed that the tumor tissue of the propranolol group was reduced, damaged, intertubular fibrous tissue infiltration, vascular endothelial cells dispersed, sparsely and wilted. Contraction, vascular wall collapse, deformation, occlusion of the lumen, destruction of the signs, while the physiological saline control group hemangioma capillary rich, dense, no lumen and vascular wall atrophy, damage signs, the CD39 light density (MD) and Ki-67 in the propranolol group were less than the normal saline control group, and the two groups showed a significant decrease. The Caspase3 and apoptosis index of propranolol group were significantly higher than that of the normal saline group (P0.05). The two groups were significantly different (P0.01), and the ELISA method was used to detect ATP concentration, which showed that the propranolol group was higher than the normal saline control group, and there was a significant difference between the two groups (P0.05). The expression of CD39 was negatively correlated with the apoptosis of the tumor cells. (r=-0.649, P0.01). Conclusion: 1. propranolol can promote the elimination of tumor in the nude mice model of hyperplastic hemangioma. The therapeutic mechanism of.2. propranolol is related to the expression of CD39 on the endothelial cells of hemangioma. It may play a cytotoxic effect of ATP by reducing the ability to reduce CD39 to ATP and thus promote the cytotoxicity of ATP. The apoptosis of the tumor cells can achieve the therapeutic effect.
【学位授予单位】：泸州医学院
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R-332;R732.2

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