普萘洛尔治疗增殖期血管瘤的实验研究

发布时间：2018-10-24 15:45

【摘要】：目的：探讨普萘洛尔对增殖期血管瘤的治疗作用及可能机制。方法：将手术切除的婴幼儿增殖期血管瘤新鲜组织块移植于裸鼠皮下，建立血管瘤动物模型；在移植后的第45天，将移植瘤组织均成活的22只实验裸鼠随机分为2组：组1(普萘洛尔治疗组)，组2(生理盐水对照组)，每组11只，分别灌注普萘洛尔溶液和生理盐水干预治疗，观察比较移植瘤的生长情况；在首次药物干预后的第15天，将实验裸鼠用颈椎脱臼法处死，切取移植的瘤体组织。免疫组化法检测移植瘤组织中血管内皮细胞MVD值，以及CD34、Ki-67、Bcl-2、VEGF的表达情况，TUNEL法检测移植瘤组织中细胞凋亡情况；酶联免疫吸附法（ELISA）检测cAMP浓度及PKA活性变化。结果：1.在药物干预后l周，普萘洛尔组与生理盐水组移植瘤体积比较，两组瘤体的体积有明显差异(P0.05)；在药物干预后2周，普萘洛尔组移植瘤体积明显小于生理盐水组，两组差异显著(P0.01)。普萘洛尔组瘤体组织颜色苍白，血供较差，，体积明显缩小，质地变硬。生理盐水组瘤体组织颜色红润，血供良好，体积无明显缩小，较有弹性。光镜下见，普萘洛尔组瘤体组织以纤维脂肪组织为主，血管内皮细胞稀疏分散，血管壁变形、塌陷，管腔结构有破坏征象；而生理盐水组瘤体组织毛细血管增生丰富，未见血管壁及管腔结构有破坏征象。2.免疫组化检测：普萘洛尔组MVD值，CD34、Ki-67、Bcl-2、VEGF的表达均明显低于生理盐水组，组间差异明显(P0.05)；TUNEL法检测普萘洛尔组细胞凋亡指数明显高于生理盐水组，两组间差异显著(P0.01)。3.酶联免疫吸附法检测：普萘洛尔组cAMP浓度和PKA活性明显低于生理盐水组，两组间差异明显(P0.05)；且普萘洛尔组cAMP浓度和PKA活性变化呈正相关(γ=0.607，P0.01)。结论：1.普萘洛尔对增殖期血管瘤有明显的治疗作用。2.普萘洛尔可诱导血管瘤内皮细胞凋亡，抑制其增殖，促进瘤体消退。3.普萘洛尔可能通过抑制cAMP／PKA信号通路对ERK／MAPK信号通路的激活，从而抑制增殖期血管瘤VEGF的分泌。
[Abstract]:Objective: to investigate the therapeutic effect of propranolol on proliferative hemangioma and its possible mechanism. Methods: the fresh tissue mass of proliferative hemangioma was transplanted into nude mice subcutaneously, and the animal model of hemangioma was established on the 45th day after transplantation. Twenty-two nude mice were randomly divided into two groups: group 1 (propranolol treatment group) and group 2 (saline control group), each group (n = 11) were treated with propranolol solution and saline respectively. At the 15th day after the first drug intervention, the nude mice were killed by cervical dislocations and the transplanted tumor tissues were removed. Immunohistochemical method was used to detect the expression of MVD and CD34,Ki-67,Bcl-2,VEGF in vascular endothelial cells, TUNEL method was used to detect apoptosis, and (ELISA) was used to detect the concentration of cAMP and the activity of PKA. The result is 1: 1. The volume of transplanted tumor in propranolol group was significantly different from that in saline group at 1 week after drug intervention (P0.05), and the volume of transplanted tumor in propranolol group was significantly smaller than that in saline group at 2 weeks after drug intervention. There was a significant difference between the two groups (P0.01). In propranolol group, the tumor tissue was pale in color, poor in blood supply, significantly reduced in volume and hardened in texture. In the saline group, the color of the tumor tissue was ruddy, the blood supply was good, the volume was not obviously reduced, and the tumor tissue was elastic. Under light microscope, the tumor tissue of propranolol group was mainly fibrous adipose tissue, vascular endothelial cells were sparsely dispersed, vascular wall was deformed, collapsed, and the structure of the lumen was damaged, while the normal saline group was rich in capillary hyperplasia. No signs of damage to vascular wall and lumen structure were found. 2. The expression of MVD and CD34,Ki-67,Bcl-2,VEGF in propranolol group was significantly lower than that in saline group, and the difference was significant (P0.05). The apoptosis index of propranolol group was significantly higher than that of normal saline group (P0.05). The difference between the two groups was significant (P0.01). Enzyme linked immunosorbent assay (Elisa) showed that the concentration of cAMP and the activity of PKA in propranolol group were significantly lower than those in saline group (P0.05), and there was a positive correlation between cAMP concentration and PKA activity in propranolol group (纬 = 0.607, P0.01). Conclusion 1. Propranolol has obvious therapeutic effect on proliferative hemangioma. 2. Propranolol can induce endothelial cell apoptosis, inhibit the proliferation of hemangioma, and promote tumor regression. Propranolol may inhibit the secretion of VEGF in proliferative hemangioma by inhibiting the activation of ERK/MAPK signaling pathway by cAMP/PKA signaling pathway.
【学位授予单位】：泸州医学院
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R739.5

【参考文献】