普萘洛尔治疗婴幼儿血管瘤临床观察及作用机制的实验研究

发布时间：2018-06-23 05:06

本文选题：婴幼儿血管瘤 + 普萘洛尔　；参考：《山东大学》2014年博士论文

【摘要】：论文纲要血管瘤(hemangioma)是婴幼儿最常见的先天性良性肿瘤,新生儿发病率2-3%,1岁以下儿童的发病率在10%左右。普萘洛尔作为非选择性β-肾上腺素受体阻滞剂,主要用于治疗心脏疾病如高血压、心绞痛、心动过速等。2008年,法国医生Leaute-Labreze C等偶然发现该药可有效抑制血管瘤的增殖,并加速瘤体消退。之后,国内外众多学者也相续将其应用于血管瘤的治疗,广泛的临床应用证实了该药对血管瘤具有明显疗效。口服普萘洛尔治疗增生期血管瘤疗效显著,同时对消退期血管瘤也有一定效果,目前已逐步取代激素成为治疗婴幼儿血管瘤的一线用药。然而,儿童口服普萘洛尔尚存在很多潜在的副作用,尤其是口服初期,需住院检测心率、血压、血糖等指标,给患儿家长及医师带来诸多不便。能否改变给药途径,采取瘤体局部外涂普萘洛尔制剂的方式,以减轻口服药物所致的潜在不良反应,鉴于此,在第一章第一部分通过对25例门诊“等待观察”的浅表型婴幼儿血管瘤患者初步尝试应用1%普萘洛尔软膏外涂治疗,结果表明疗效显著且未发现可检测到的与外用普萘洛尔软膏相关的不良反应。此外,口服普萘洛尔治疗血管瘤,国内外文献较多集中在对疗效评估,而对服药期间因治疗所致的副作用或不良反应发生情况研究尚存在不足。故在本章第二部分,回顾性分析近年来我科接受口服普萘洛尔治疗的106例血管瘤患儿,以评估药物治疗相关的副作用或不良反应发生情况,为用药安全提供依据。 Folkman等学者认为,血管瘤是一种血管形成性质疾病,失控的血管生成是其增殖的主要因素。该过程包括了内皮细胞增殖、细胞外基质的选择性降解、细胞的迁移以及血管通透性的增加。细胞生长因子是一类能诱导或促进新血管形成的物质,可调控内皮细胞的增殖,影响血管形成,对血管瘤的发生发展起关键作用。类表皮生长因子域7(EGFL7)是一种内皮细胞特异性分泌因子,选择性地表达于肿瘤新生血管和增生性组织中,在血管生成过程中起关键作用。由此我们提出,①EGFL7是否在血管瘤组织中表达及在该病发生发展的不同时期是否存在差异表达?②普萘洛尔对血管瘤显著的临床疗效是否通过抑制EGFL7表达?上述问题是进一步研究普萘洛尔对血管瘤作用机制所要解决的问题。鉴于此,第二章第一部分利用免疫组织化学染色技术检测类表皮生长因子域7(EGFL7)在婴幼儿血管瘤组织中的表达,并探讨其在血管瘤发生发展过程中的作用和意义。本章第二部分,通过检测普萘洛尔干预下人脐静脉内皮细胞(HUVEC)增殖、成管、凋亡及EGFL7的表达影响,初步探讨普萘洛尔抑制血管瘤增生的作用机制。第一章普萘洛尔治疗婴幼儿血管瘤临床观察第一部分普萘洛尔外用软膏治疗浅表型婴幼儿血管瘤疗效评估目的探讨局部外涂普萘洛尔软膏治疗浅表型婴幼儿血管瘤的临床疗效及安全性。方法本组共25例(共28处血管瘤,女21例,男4例,年龄1月至10月,平均4个月)浅表型婴幼儿血管瘤患儿,门诊接受瘤体表面外涂1%普萘洛尔软膏,每日3次,均匀涂于瘤体表面,时间5周至59周(平均21周),依据服药前后瘤体表面张力、大小、颜色变化,按照显著有效、部分有效、无效3级评分法对近期疗效进行评价。同时,观察用药后副作用以评估用药安全性。结果28处血管瘤中,16(57%)例显著有效；9(33%)例部分有效；3(10%)例为无效。1%普萘洛尔软膏对浅表型婴幼儿血管瘤的总有效率为90%(95%可信区间72%-98%)。所有患儿均未出现可检测到的不良反应。结论外涂普萘洛尔软膏能有效促进浅表型婴幼儿血管瘤的消退,且未发现口服普萘洛尔常见的不良反应,可以作为婴幼儿血管瘤随访观察期间的一种安全、有效辅助治疗手段。第二部分口服普萘洛尔治疗婴幼儿血管瘤不良反应观察目的：观察口服普萘洛尔治疗婴幼儿血管瘤不良反应,评估其用药安全性。方法：2009年9月至2013年11月间山东省立医院烧伤整形美容科口服普萘洛尔治疗的106例婴幼儿血管瘤患儿,女性71例,男性35例,年龄1-14个月(平均5.1个月)。服药剂量每日1-1.5mg/kg,分3次给药,随访2-10个月,期间对出现的用药相关副作用或并发症进行安全性评估。结果：106例患儿服药期间及随访阶段瘤体均得到有效控制且有程度不同的消退。18例病人(17%,18/106,4例并发两种以上副作用)出现与用药相关的副作用,其中10(9.4%)例口服期间发生腹泻,7(6.6%)例出现一过性血压下降,2(1.9%)例溢奶,2(1.9%)例夜间睡眠差,易激和四肢末端发凉各1例(0.94%)。结论：小剂量(1-1.5mg/kg.day)、分次口服普萘洛尔治疗婴幼儿血管瘤疗效显著,且不良反应发生率低,用药安全,可作为婴治疗幼儿血管瘤的用药选择。第二章普萘洛尔抑制婴幼儿血管瘤增殖的机制探讨第一部分类表皮生长因子域7(EGFL7)在婴幼儿血管瘤组织中的表达及意义目的：检测类表皮生长因子域7(EGFL7)在不同时期婴幼儿血管瘤组织中的表达情况并探讨其在该病发生发展过程中的作用及意义。方法：收集2007年5至2013年9月山东大学附属省立医院烧伤整形美容科手术切除组织病理诊断为婴幼儿血管瘤的石蜡标本49例(男16例,女33例),依据Mulliken标准将标本分成增生期组、退期组、消退完成期组,以正常皮肤作为对照组。应用免疫组织化学法对各期婴幼儿血管瘤组织及正常皮肤中的类表皮生长因子域7(EGFL7)表达情况进行检测,利用计算机图像分析技术分别测量其平均光密度。结果：EGFL7阳性反应定位于血管瘤细胞胞浆中,在血管瘤增生期强阳性或阳性表达,消退期阳性表达,消退完成期和正常皮肤中弱阳性或阴性性表达。结论：EGFL7与增生期婴幼儿血管瘤关系密切,参与了婴幼儿血管瘤病理变化过程,在该病增生期和消退早期细胞增殖和血管形成阶段起重要作用。第二部分普萘洛尔对人脐静脉内皮细胞(HUVEC)生物学行为及EGFL-7表达的影响目的：探讨普萘洛尔对人脐静脉内皮细胞增殖、成管能力、凋亡及EGFL-7表达影响,为进一步研究普萘洛尔治疗婴幼儿血管瘤机制提供理论依据。方法：以离体的人脐静脉内皮细胞为实验对象,采用MTT法、成管实验、流式细胞仪测定不同浓度的普萘洛尔(0μmol/L、25μmo1/L、50μmol/L、100μmo1/L、125μmol/L、150μmol/L、175μmol/L、200μmol/L)对HUVEC增殖、成管能力、凋亡的影响及RT-PCR、western blot法测定类表皮生长因子域-7的变化。结果：普萘洛尔药物干预24和48小时后,药物浓度为≥25um时对人脐静脉内皮细胞增殖能力的抑制率与正常对照组相比有统计学意义(P0.01),且在25μmol/L、50μmol/L、100μmol/L、125μmol/L浓度下抑制率成明显上升趋势；流式细胞仪检测不同浓度普萘洛尔实验组和空白组对比(24小时),细胞凋亡率逐步增加；药物处理组24h细胞成管个数与阴性对照无统计学差异,48小时对成管能力有一定抑制作用；RT-PCR和Western blot实验结果显示药物处理组与空白组对比,人脐静脉内皮细胞的EGFL-7表达呈现浓度性抑制作用,有显著差异(P0.05)。结论：普萘洛尔抑制人脐静脉内皮细胞的增殖,加速细胞早期凋亡,两者呈现浓度依赖性,对成管能力有一定抑制作用,呈现时间依赖性；普萘洛尔成浓度依赖性抑制人内皮细胞中EGFL-7的表达,由此推测普萘洛尔抑制增生期血管瘤作用机制与通过抑制EGFL-7介导的促血管生成作用,最终达到加速血管瘤消退。
[Abstract]:Thesis proposal
Hemangioma (hemangioma) is the most common congenital benign tumor in infants. The incidence of newborns is 2-3% and the incidence of children under 1 years old is about 10%. Propranolol is used as a non selective beta adrenergic receptor blocker, which is mainly used in the treatment of heart diseases such as hypertension, cardiac tachycardia, tachycardia, and other.2008 years, and the French doctor Leaute-Labreze C It is found that the drug can effectively inhibit the proliferation of hemangioma and accelerate the decline of the tumor body. After that, many scholars at home and abroad also continue to apply it to the treatment of hemangioma. Extensive clinical application confirms that the drug has obvious curative effect on hemangioma.
Oral propranolol has a significant effect on the treatment of hyperplastic hemangioma and also has some effect on the hemangioma in the subsiding period. At present, the hormone has gradually replaced the hormone as a first-line drug for the treatment of infantile hemangioma. However, there are many potential side effects in children's oral propranolol, especially in the early stage of oral administration, which need to be hospitalized to detect heart rate, blood pressure, blood sugar and so on. The index can bring many inconvenience to the parents and doctors. Can we change the way of drug delivery, take the way of applying propranolol to reduce the potential adverse reactions caused by oral medicine. In the first chapter, the first part of the first chapter is to try the initial attempt of 25 cases of superficial infantile hemangioma "waiting for observation" in the outpatient clinic. The treatment with 1% propranolol ointment showed significant effect and no detectable adverse reactions associated with the external propranolol ointment. In addition, the oral propranolol was more concentrated in the treatment of hemangioma at home and abroad, and the side effects or adverse reactions caused by treatment during the medication were studied. In the second part of this chapter, a retrospective analysis of 106 cases of hemangioma treated by oral propranolol in recent years was reviewed to assess the side effects or adverse reactions related to drug treatment, and to provide a basis for the safety of drug use.
Folkman and other scholars believe that hemangioma is a angiogenic disease. Out of control angiogenesis is a major factor in its proliferation. This process includes endothelial cell proliferation, selective degradation of extracellular matrix, cell migration, and increased vascular permeability. Cell growth is a class of substances that can induce or promote new vascular formation. Cytoplasm, which regulates the proliferation of endothelial cells and affects angiogenesis, plays a key role in the development of angioma. The epidermal growth factor domain 7 (EGFL7) is a specific secretory factor of endothelial cells, which is selective in neovascularization and proliferative tissue of tumor, and plays a key role in the process of blood Guan Shengcheng. Therefore, we propose, (1) EGFL Is there any difference in expression of 7 in hemangioma tissue and in different periods of the development of the disease? 2. Is the significant clinical effect of propranolol on hemangioma by inhibiting EGFL7 expression? The above question is a further study of the problem of propranolol's mechanism of action on hemangioma. In view of this, the first part of the second chapter is beneficial. Immunohistochemical staining was used to detect the expression of epidermal growth factor domain 7 (EGFL7) in the tissue of infantile hemangioma, and to explore its role and significance in the development of hemangioma. The second part of this chapter is to detect the proliferation, tube, apoptosis and EGFL7 expression of human umbilical vein endothelial cells (HUVEC) under the intervention of propranolol. The mechanism of propranolol inhibiting hemangioma proliferation was preliminarily explored.
Chapter 1 clinical observation of propranolol in treatment of infantile hemangioma
Part I: propranolol external ointment in the treatment of superficial infantile hemangioma
Objective to investigate the clinical efficacy and safety of topical propranolol ointment in the treatment of superficial infantile hemangioma.
Methods a total of 25 cases (28 cases of hemangioma, 21 women, 4 men, 4 men, from January to October, 4 months, average 4 months) were treated with 1% propranolol ointment on the surface of the tumor, 3 times a day, evenly coated on the surface of the tumor for 5 to 59 weeks (21 weeks flat), according to the surface tension, size and color changes of the tumor before and after the medication. The short-term efficacy was evaluated according to the significant, effective, invalid and 3 grade scoring method. Meanwhile, the side effects of medication were observed to evaluate the safety of medication.
Results of the 28 hemangiomas, 16 (57%) cases were significantly effective, 9 (33%) were partially effective, and 3 (10%).1% propranolol ointment had a total effective rate of 90% (95% confidence interval 72%-98%) for superficial infantile hemangioma. All the children had no detectable adverse reactions.
Conclusion the application of propranolol ointment can effectively promote the decline of superficial infantile hemangioma, and not found the common adverse reaction of oral propranolol. It can be used as a safe and effective adjuvant therapy for the follow-up observation of infantile hemangioma.
The second part of oral propranolol treatment of infantile hemangioma adverse reactions observed
Objective: To observe the adverse reaction of propranolol in the treatment of infantile hemangioma and evaluate its safety.
Methods: from September 2009 to November 2013, 106 children with infantile hemangioma treated by oral propranolol, the Department of burn and plastic surgery, Shangdong Province-owned Hospital, were treated with 106 cases of infant hemangioma, 71 women, 35 males, 1-14 months of age (average 5.1 months). The dosage of the medicine was given for 2-10 months, and the drug related side effects were observed for 2-10 months. A safety assessment of the disease.
Results: 106 cases of.18 patients were effectively controlled and varying degrees of tumor withdrawal during and follow-up period (17%, 18/106,4 cases with more than two side effects) associated with the side effects, including 10 (9.4%) cases of oral diarrhea, 7 (6.6%) cases of transient blood pressure decline, 2 (1.9%) cases of spilled milk, 2 (1.9%) cases. Night sleep was poor, irritability and cold extremities were found in 1 cases (0.94%).
Conclusion: small dose (1-1.5mg/kg.day) and oral propranolol are effective in the treatment of infantile hemangioma, and the incidence of adverse reactions is low. The drug is safe and can be used as a choice for infant hemangioma.
The second chapter is the mechanism of propranolol inhibiting the proliferation of infantile hemangioma.
Expression and significance of epidermal growth factor domain 7 (EGFL7) in infantile hemangioma tissues
Objective: to detect the expression of epidermal growth factor domain 7 (EGFL7) in the tissues of infantile hemangioma at different times and to explore its role and significance in the development of the disease.
Methods: 49 paraffin specimens (16 males and 33 females) diagnosed as infantile hemangiomas were collected from 5 to September 2013 of Shandong University from 5 to September 2013 in 2007. According to the standard, the specimens were divided into the hyperplastic period group, the retreating group, the subsiding completion period, and the normal skin as the control group. Histochemical method was used to detect the expression of epidermal growth factor domain 7 (EGFL7) in the tissues of infantile hemangioma and normal skin, and the average optical density was measured by computer image analysis.
Results: the positive reaction of EGFL7 was located in the cytoplasm of hemangioma cells. The positive or positive expression of the hemangioma was positive or positive, the positive expression in the subsiding stage, the fading phase and the weak positive or negative expression in the normal skin.
Conclusion: EGFL7 is closely related to the hyperplastic infantile hemangioma, and participates in the pathological changes of the infantile hemangioma, which plays an important role in the stage of proliferation and the stage of angiogenesis and angiogenesis in the early stage of the disease.
The second part of propranolol affects the biological behavior and EGFL-7 expression of human umbilical vein endothelial cells (HUVEC).
Objective: To investigate the effect of propranolol on the proliferation, tubular capacity, apoptosis and EGFL-7 expression of human umbilical vein endothelial cells, and to provide a theoretical basis for the further study of propranolol in the treatment of infantile hemangioma.
Methods: using the isolated human umbilical vein endothelial cells as the experimental object, the MTT method was used to test the proliferation of propranolol (0, 25, 50, 50, 50, 100, 125 u mol/L, 150 mol/L, 175, mol/L, 200 micron) for HUVEC colonization, the effect of tubular capacity, apoptosis and RT-PCR, Western assay The changes in the epidermal growth factor domain -7.
Results: after 24 and 48 hours of propranolol intervention, the inhibition rate of the proliferation ability of human umbilical vein endothelial cells was significantly higher than that of the normal control group (P0.01), and the inhibition rate of the human umbilical vein endothelial cells was significantly higher than that of the normal control group (P0.01), and the inhibition rate of the human umbilical vein endothelial cells was significantly increased at 25 u mol/L, 50 mu, 100 mu mol/L, and 125 u mol/L concentration; the flow cytometry was used to detect the different concentrations. The apoptosis rate of the propranolol experiment group and the blank group (24 hours) increased gradually. The number of 24h cells in the drug treatment group was not statistically different from that of the negative control. 48 hours had a certain inhibitory effect on the ability of the tube formation. The results of RT-PCR and Western blot showed that the drug treatment group was compared with the blank group, and the EG of the human umbilical vein endothelial cells was EG. FL-7 expression showed a significant inhibitory effect (P0.05).
Conclusion: propranolol inhibits the proliferation of human umbilical vein endothelial cells and accelerates the early apoptosis of the cells. Both of them have a concentration dependence and have a certain inhibitory effect on the ability of tube formation. Propranolol inhibits the expression of EGFL-7 in human endothelial cells, thus suppresses the effect of propranolol to inhibit the proliferation of hemangioma. The mechanism can ultimately accelerate the regression of hemangioma by inhibiting EGFL-7 mediated angiogenesis.
【学位授予单位】：山东大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R732.2

【参考文献】