【作者】 习明；

【导师】 胡卫列；

【作者基本信息】 南方医科大学 ， 泌尿外科， 2008， 硕士

【摘要】 目的:为了探讨CGRP(Calcitoni rgene related peptide,降钙素基因相关肽)、SP(substance P,P物质)、NPY(Neurpeptide Y,神经肽Y)在肾上腺疾病发生过程中的所起的作用及临床意义,用免疫组化的方法,通过对肾上腺皮质增生、皮质腺瘤、髓质增生、嗜铬细胞瘤和正常肾上腺等不同组织,进行CGRP、SP、NPY免疫组化染色,分别观察其在肾上腺疾病组织中的分布特点。方法:选取CGRP、SP、NPY作为标记物,运用免疫组化方法对广州军区总医院1980年12月～2006年12月提供的常规石蜡包埋组织块,其中肾上腺皮质增生、皮质腺瘤、髓质增生、嗜铬细胞瘤和正常肾上腺各15例,分别进行CGRP、SP、NPY染色。新鲜标木经10%福尔马林固定18-24h,常规石蜡包埋,连续切片厚5μm,1张作HE染色,其余分别作NPY、CGRP、SP免疫组化染色。正常肾上腺作为阳性对照,PBS代替一抗作为阴性对照。阳性物均位于胞浆内,呈棕黄色。判断标准:阳性细胞面积占细胞总面积的百分比。比较肾上腺皮质增生、皮质腺瘤、髓质增生、嗜铬细胞瘤和正常肾上腺等不同病例皮质和髓质中NPY、CGRP、SP染色阳性细胞占细胞总数百分比之间的差异。统计数据用SPSS13.0处理,CGRP、SP、NPY阳性细胞面积数据行完全随机设计方差分析,CGRP、SP、NPY阳性细胞面积与患者入院血压行直线相关分析,检验水准均为α=0.05。结果:CGRP、SP在正常肾上腺髓质、肾上腺髓质增生、嗜铬细胞瘤表达呈阴性,而在正常肾上腺皮质、肾上腺皮质增生、肾上腺皮质腺瘤均有不同程度的表达肾上腺皮质腺瘤、皮质增生、正常肾上腺皮质中均可见CGRP、SP染色阳性细胞,三者CGRP、SP染色面积有显著差异(P=0.002,P=0.006),其中肾上腺皮质腺瘤阳性细胞面积显著高于皮质增生、正常肾上腺皮质,肾上腺皮质增生表达高于正常肾上腺皮质;并且CGRP、SP阳性细胞面积与肾上腺皮质腺瘤、肾上腺皮质增生患者的血压存在正相关关系(相关系数分别为r=0.801,P=0.010;r=0.686,P=0.021;r=0.811,P=0.022;r=0.523,P=0.013)。NPY在正常肾上腺皮质、肾上腺皮质增生、肾上腺皮质腺瘤表达呈阴性,肾上腺髓质增生、肾上腺嗜铬细胞瘤和正常肾上腺髓质中均可见NPY染色阳性细胞,三者NPY染色面积有显著差异(P=0.018),其中肾上腺嗜铬细胞瘤阳性细胞面积显著高于肾上腺髓质增生和正常肾上腺髓质,肾上腺髓质增生表达高于正常肾上腺髓质;并且NPY阳性细胞面积与肾上腺嗜铬细胞瘤、肾上腺髓质增生患者的血压存在正相关关系(相关系数分别为r=0.856,P=0.031;r=0.792,P=0.009)。结论:1.CGRP、SP与肾上腺皮质疾病患者高血压有密切相关,对肾上腺皮质疾病激素的分泌可能起到到促进作用。NPY与肾上腺嗜铬细胞瘤、肾上腺髓质增生患者的血压密切相关,可能促进肾上腺髓质的激素分泌。2.CGRP、SP和NPY在肾上腺疾病的发生发展中可能具有重要意义,由正常肾上腺组织向肾上腺增生和肾上腺腺瘤或肾上腺嗜铬细胞瘤的发展过程中CGRP、SP与NPY经历了由低表达到高表达的过程。这可能是治疗和诊断肾上腺疾病新的靶点。3.以往的文献报道已经证明NPY由肾上腺嗜铬细胞生成分泌,并且NPY介导了直接的血管收缩作用;而我们的实验也已证明NPY在肾上腺嗜铬细胞瘤、肾上腺髓质增生疾病中是高表达的,所以我们推测NPY可能是肾上腺嗜铬细胞瘤、肾上腺髓质增生疾病中,除了NA(noradrenaline)和AD(adnephrin)之外,分泌的又一外周缩血管物质。更多还原

【Abstract】 OBJECTIVE:For reveal the the roles and clinical significance of Neurpeptide Y (NPY),substance P(SP) and Calcitoni rgene related peptide(CGRP) in the in adrenal gland,we study the distribution of NPY,CGRP,SP in adrenal gland by immunohistochemistry and statistics percentage of colour positive cell to compare difference among adrenocortical adenoma,adrenocortical hyperplasia, pheochromocytoma,adrenal medullary hyperplasia and normal adrenal.METHODS:To collect 75 conventional paraffin wax tissues of adrenocortical adenoma, adrenocortical hyperplasia,pheochromocytoma,adrenal medullary hyperplasia and normal adrenal from our hospital from December 1980 toDecember 2006 and normal adrenal corned from corpse.Every group is 15 cases.Immunohistochemical technique was applied to detect the expression of NPY,CGRP and SP in routinely processed tissue specimens from human adrenal neoplasm and hyperplasia.All tissues were fixed by formalin,its concentration is 10 percentage.Fixed time is between eighteen hours and twenty-four hours.Specimens were embedded by ozocerite and chip to Sum thickness.One specimen was HE stain,another was NPY, CGRP,SP immunoreactivity.Normal adrenal was positive contrast and PBS replaced first antibody was negative contrast.Positive matter was found in cytolymph and its colour was brown.Judgement standard is percentage of positive cell.The destination was to study the characteristic of NPY,CGRP and SP in adrenal by immunohistochemistry and statistics percentage of colouer positive cell to compare difference to adrenocortical adenoma adrenocortical hyperplasia, pheochromocytoma,adrenal medullary hyperplasia and normal adrenal.RESULTS:CGRP,SP immunoreactivity was regularly seen in adrenocortical adenoma, adrenocorticohyperplasia and normal adrenal cortex and there was a significant difference among the three(P=0.002,P=0.006),but neither in the pheochromocytoma,adrenal medullary hyperplasia nor in the normal adrenal medulla.CGRP,SP expression was higher in adrenocortical adenoma tissue than that of adrenocorticohyperplasia and normal cortex,the same with SP.Besides,higher CGRP,SP expression were present in adrenocorticohyperplasia group than those of normal adrenal cortex group.There also was a positive correlation between CGRP,SP positive cell and blood pressure in adrenocortical adenoma and adrenocorticohyperplasia(r=0.801,P=0.010;r=0.686,P=0.021;r=0.811,P=0.022; r=0.523,P=0.013).NPY positive cell were showed in pheochromocytoma,adrenal medullary hyperplasia and normal adrenal medulla and there was a significant difference among the three(P=0.018),but not both in the adrenocortical adenoma, adrenocorticohyperplasia and normal adrenal cortex.NPY expression was higher in pheochromocytoma tissue than that of adrenal medullary hyperplasia and adrenal medulla.Besides,higher NPY expression were present in adrenal medullary hyperplasia group than those of normal adrenal medulla group.There also was a positive correlation between NPY positive cell and blood pressure in adrenal medullary hyperplasia and adrenal Pheochromocytoma(r=0.856,P=0.031;r=0.792, P=0.009).CONCLUSIONS:1.Maybe CGRP and SP correspond to induces adrenal hormone release Besides,there was a positive correlation between CGRP,SP positive cell and blood pressure in adrenocortical adenoma and adrenocorticohyperplasia.The same with NPY for pheochromocytoma and adrenal medullary hyperplasia.2.CGRP,SP and NPY may play an important role in the and the development of adrenal disease.The expression of CGRP,SP and NPY change from low to high in the process from normal adranal to adrenal hyperplasia and adrenal tumor.CGRP,SP and NPY are functional and may represent the molecular basis of adrenal Pheochromocytoma,as well as a potential new target for the diagnosis and therapy of adrenal disease.3.Previous literature has proved that NPY secrete from adrenal chromaffin cell and our experiment also has proved NPY high expression in adrenal medulla disease,so, we think that maybe NPY is the another vasoconstriction substance except norepinephrine and adnephrin,which pheochromocytoma or adrenal medullary hyperplasia secreted.更多还原