人白细胞介素24mRNA在瘢痕疙瘩中的表达及意义  

Expression and clinical significance of interleukin-24 mRNA in keloid

作　　者：李建赤[1] 梁杰[1] 罗少军[1] 张培华[1] 彭智[1] 李平[1] 

机构地区：[1]广东医学院整形外科研究所,广东省湛江市524001

出　　处：《中国组织工程研究与临床康复》2007年第32期6432-6435,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：广东省自然科学基金资助(06028957)~~

摘　　要：目的:从基因水平测量瘢痕疙瘩组织中白细胞介素24的表达水平,探讨白细胞介素24在瘢痕疙瘩发生、发展过程中的作用和意义。方法:实验于2005-10/2006-09在广东医学院整形外科研究所完成。①选取2004-06/2005-10广东医学院附属医院整形外科收治的患者,瘢痕疙瘩标本12例,正常瘢痕标本10例,行巨乳缩小、除皱术、植皮等患者正常皮肤标本12例,患者均知情同意且自愿捐献标本,实验经医学伦理委员会批准。标本取材部位为颜面、前胸、四肢等,切取后液氮保存。②低温条件下切取秤量组织,采用Trizol法提取总RNA。电泳鉴定总RNA完整性,并统一调整总RNA含量为10g/L,-70℃储存。RT-PCR二步法合成cDNA。③以正常皮肤、正常瘢痕为对照,以GAPDH作为扩增内参照基因,将正常皮肤、正常瘢痕和瘢痕疙瘩各类标本总RNA反转录的cDNA模板浓度调整相对一致进行扩增反应。以白细胞介素24mRNA与GAPDHmRNA的光密度积分值之比作为各类组织标本中白细胞介素24的相对含量,比较白细胞介素24mRNA在正常皮肤、正常瘢痕及瘢痕疙瘩组织中的表达情况。结果:①各类组织标本中总RNA抽提结果:正常皮肤、正常瘢痕和瘢痕疙瘩组织中抽提总RNA经甲醛变性凝胶电泳后显示较清晰的18s和28s条带,经紫外分光光度计测定A260/A280≈2.0。②各类组织标本中白细胞介素24mRNA的表达:白细胞介素24和GAPDH基因表达产物通过RT-PCR方法得到的特异性DNA片段长度分别为173bp和577bp。瘢痕疙瘩的白细胞介素24mRNA与GAPDHmRNA的吸光度比值明显低于正常皮肤、正常瘢痕(0.577±0.113,1.070±0.185,1.139±0.195;t=7.436×10-8～4.745×10-8,P均<0.01),正常皮肤与正常瘢痕白细胞介素24mRNA的相对表达量基本一致(t=0.405,P>0.05)。结论:瘢痕疙瘩的形成可能与白细胞介素24在组织中的表达降低有关。提示采用基因疗法提高早期瘢痕疙瘩中白细胞介素24的含量�AIM： To investigate the expression of interleukin 24 （IL-24） in keloid at the gene level, and to determine its effects and significance in the formation and development of keloid. METHODS： The experiment was conducted in the Research Institute of Plastic Surgery of Guangdong Medical College from October 2005 to September 2006. ①The specimens including 12 of keloid, 10 of normal scars, and 12 of normal skins, which were excised from the patients who underwent reduction mammaplasty, rhytidectomy or dermatoplasty, were collected from Department of Plastic Surgery, Affiliated Hospital of Guangdong Medical College from June 2004 to October 2005 including. All patients were informed the experiment purpose and voluntarily donated the specimens. Meanwhile, the experiment was approved by the Hospital Ethics Committee. The specimens were harvested from face, prothorax, limbs, etc, and were then preserved in liquid nitrogen.② The tissues were cut and weighed under hypothermal condition. Total RNA was then extracted by Trizol method. The integrity of total RNA was identified by electrophoresis. The concentration of total RNA was adjusted to 10 g/L, and then stored at -70 ℃ freezer, cDNA was synthesized by two step method of RT-PCR. ③With the normal skin and normal scar as control and the GAPDH as amplified intra-reference gene, the cDNA, which were synthesized by reverse transcription of total RNA of all kinds of specimens was undertook PCR with the concentration were adjusted to conformity relatively. With the ratio of photodensity score of IL-24 mRNA and GAPDH as the relative amount of IL-24 in all kinds of collections, the expression of IL-24 in normal skin, normal scar and keloid were compared. RESULTS： ①The total RNA of normal skin, normal scar and keloid displayed clear bands of 18 s and 28 s by formaldehyde denaturing gel electrophoresis, while A260/A280 was about 2.0 which was determined by ultraviolet spectrophotometer. ② The specific DNA fragment length of the gene expression product of IL

关 键 词：瘢痕 瘢痕疙瘩 人白细胞介素24 基因表达 

分 类 号：R619.6[医药卫生—外科学]