【作者】 刘文；

【导师】 霍继荣；

【作者基本信息】 中南大学 ， 内科学， 2008， 硕士

【摘要】 目的:食管静脉曲张破裂出血是肝硬化门脉高压症的严重并发症之一,病死率高。采用内镜下硬化注射治疗曲张静脉破裂出血是目前临床上主要抢救措施。市场上硬化剂种类繁多,为了解不同硬化剂的疗效、副作用及以及重复治疗的时机,本研究利用不同硬化剂注射动物静脉血管及周围组织的组织标本,通过免疫组化评价VEGF和CD34在相应血管及组织的表达,观察不同硬化剂注射对血管及周围组织的血管、组织修复的影响,为临床上选择相对理想的硬化剂、重复治疗时机提供理论依据。方法:不同硬化剂注射动物静脉血管及周围组织的组织标本来源于前课题组完成的动物实验。动物实验选择体重约为2—3Kg的新西兰兔,雌雄不限,随机分成5组,分别注射1%乙氧硬化醇、无水酒精、5%鱼肝油酸钠、凝血酶以及生理盐水对照组。各实验组分为静脉内注射组、静脉内和静脉旁联合法注射组。取注射不同硬化剂后的耳缘静脉血管及周围组织作为标本。分别为注药后第1天、第3天、第7天、第14天处死实验动物,对不同硬化剂注射的静脉血管及周围组织进行常规HE染色以及免疫组化法检测VEGF和CD34的表达,评估不同硬化剂的疗效、副作用、及组织修复时间。结果:HE染色显示三种硬化剂注射后血管内皮及周围组织损伤有一定程度损伤,其中无水酒精最重,鱼肝油酸钠次之,乙氧硬化醇最轻,而凝血酶组与对照的生理盐水组无明显差异。VEGF采用积分评判,CD34采用MVD计数评价,两者在乙氧硬化醇组表现为明显升高的趋势,第3天达到高峰,第7-14天逐渐下降至正常,两种注射方式间无明显差异(VEGF为P=0.199,MVD为P=0.632),各时段间对比P＜0.05;VEGF和MVD在无水酒精组表现为明显下降趋势,到第7天和第14天表达几乎为零,两种注射方式之间对比有明显差异(VEGF和MVD均P＜0.05),联合注射方式VEGF和MVD更低,各时段之间对比P＜0.05;VEGF和MVD在鱼肝油酸钠组均表现为轻度上升继而明显下降的趋势,两种注射方式之间无明显差异(VEGF为P=0.184,MVD为P=0.549),各时段之间对比P＜0.05;凝血酶组VEGF和MVD表现无明显变化,两种注射方式间无明显差异(VEGF为P=0.913,MVD为P=0.256),各时段间亦无差异(VEGF为P=0.069,MVD为P=0.08);VEGF评分和MVD计数在各硬化剂组间以及与凝血酶和生理盐水组之间两两对比均有明显差异,P值均小于0.05;而凝血酶与生理盐水组之间对比无明显差异,P值大于0.05。结论:不同硬化剂注射血管后动物组织学标本HE染色及VEGF和CD34的表达提示;HE染色显示1%乙氧硬化醇具有对血管组织损伤小、血栓形成快、组织恢复快,VEGF和CD34的表达在3天达高峰,两周后恢复正常,组织学及VEGF和CD34的表达一致,显示1%乙氧硬化醇是一种有效、组织损伤小、修复快、副作用少的理想的硬化剂,重复治疗周期为1周左右;HE染色显示无水酒精对血栓形成快有效,但组织损伤大,修复慢,VEGF和CD34表达第3天明显下降,第7—14天几乎不表达,与组织学表现一致,提示组织损伤大,修复受抑制,建议临床上不用无水酒精作为常规的食管静脉曲张治疗的硬化剂,由于组织修复缓慢,重复治疗周期应在2周以上。HE染色显示5%鱼肝油酸钠同样具有较好的栓塞效果,对血管及周围组织损伤较1%乙氧硬化醇严重,VEGF和CD34表达表现为轻度上升后再明显下降的过程,组织修复介于1%乙氧硬化醇和无水酒精之间,临床可以选用,重复治疗周期应在1-2周;凝血酶注射后和生理盐水对照组一样,不产生无菌性炎症,不形成血栓,VEGF和CD34表达无变化,提示凝血酶无硬化效果,不可作为硬化剂使用。更多还原

【Abstract】 Aims : Esophageal varices bleeding is a serious complication of hepatic cirrhosis portal hypertension , its dangerous condition. endoscopic variceal sclerotherapy was the major rescue measures for treatment of esophageal varices bleeding. Sclerosing agent have a great variety in market, in order that the efficacy, side effects and repeat times for treatment after different sclerosing agent injection, this study utilize the animal vein and surrounding tissue specimens after different sclerosing agent injection, through the immunohistochemical evaluatation of VEGF and CD34 in the scorresponding expression of vascular and organizations, to observe the impact of blood vessels and surrounding tissue after different sclerotherapy, and tissue repair, so as to the choice of relatively ideal scleroing agent, repeat the treatment time to provide a theoretical basis.Methods: The different sclerotherapy animal vein and the surrounding tissue specimens from the completed animal experiment. Select weight of about 2-3Kg of rabbits whose sex was no limited, were randomly divided into five groups, 1% aethoxysklerol group （n=8）, absolute alcohol group （n=8）, 5% sodium morrhuate group （n=8）, thrombin group （n=8）, normal sodium control group （n=4）. Every experimental group was separated into the intravenous injection group, the other was the intravenous and paravenous joint injection group. Select the ear vein and surrounding tissue after different scleroing agent injection. In 1^st day, 3^rd days, 7^th days, 14^th days after injection laboratory animals were sacrificed for different sclerosing agent injection group, the veins and surrounding tissue were carry out conventional HE staining and immunohistochemistry detected expression of VEGF and CD34, to evaluate the curative effects, side effects, and tissue repair time in different group.Results: HE staining showed that the vascular endothelial and surrounding tissue after three sclerosing agent injection have a varying degree injury, of which the most absolute alcohol, followed sodium morrhuate, aethoxysklerol lightest, and thrombin group and the normal sodium control group was no significant difference. VEGF using integral evaluation, CD34 use MVD count evaluation, both in the aethoxysklerol group showed significantly higher trend, the 3^rd day reached a peak, 7^th -14^th day decreased gradually to normal, two injecting style no obvious differences （VEGF for P=0.199, MVD for P=0.632）, the contrast between the periods P<0.05; VEGF and MVD in the absolute alcohol group showed significantly downward trend, to 7^th and 14^th day almost zero, comparison between the two injecting style have a significant difference （VEGF and MVD both P<0.05）, the joint injection lower, the contrast between the periods P<0.05; VEGF and MVD in sodium morrhuate group performances for the light increased then obvious downward trend, there are no significant difference between two injection （VEGF for P=0.184,MVD for P=0.549）, the contrast the periods P<0.05; In thrombin group, VEGF and MVD no significant difference （VEGF for P=0.913,MVD for P=0.256）, there is no difference between the various time slots （VEGF for P=0.069,MVD for P=0.08）; VEGF score and MVD count contrast in the sclerosing agent groups and anyone with thrombin or normal sodium control group have significant differences, all P<0.05; There was no significant contrast between thrombin and normal sodium, P>0.05.Conclusions: According to pathological changes seen from HE staining, as well as the expression of VEGF and CD34 in animal tissue specimens after different scleroing agent injection, HE staining showed 1% aethoxysklerol have some characteristics, such as blood vessels and tissue injury small, thrombosis fast and organizations recovery quick. VEGF and CD34 expression in 3^rd day reach summit, two weeks later return to normal, confirmed that it is a more ideal sclerosing agent, but the price is more expensive, proposed joint injection treatment in larger vessels, the treatment cycle about 1 week; HE staining showed Absolute alcohol have some feature that thrombosis fast , tissue injury large and tissur repair slow, VEGF and CD34 expression were consistent, in 3^rd day decreased significantly, 7^th - 14^th day almost no expression, suggesting that its should not be used as clincical application of conventional sclorsing agent to treatment esophageal varices, even if we use, it should be to the mian intravenous injection, the treatment cycle should be over in two weeks. HE staining showed 5% sodium morrhuate have good embolization results, but the blood vessels and surrounding tissue injury more serious than aethoxysklerol, VEGF and CD34 expression showed the process of mild rise then significantly decreased, tissue repair between aethoxysklerol and absolute alcohol, but its low price, there is still space for clinical application, the joint injection style was recommendation, the treatment cycle in one week above. Thrombin injection and normal sodium control group have same effect, no aseptic inflammation, thrombosis can not be formed , VEGF and CD34 expression no change, don’t have to use as a sclerosing agent.更多还原