表皮松解性角化過度型魚鱗病一家系KRT10基因突變的研究及應(yīng)用維甲酸治療的臨床觀察
發(fā)布時(shí)間：2018-11-26 17:22
【摘要】： 前言 魚鱗病(ichthyosis)是一組以皮膚干燥伴片層魚鱗狀粘著性鱗屑為特征的角化異常性遺傳性皮膚病。根據(jù)遺傳機(jī)制、臨床表現(xiàn)及組織病理學(xué)的差異,主要分為四型：尋常型魚鱗病(ichthyosis vulgaris, IV )、表皮松解性角化過度型魚鱗病(epidermolytic hyperkeratosis, EHK)、X連鎖型魚鱗病(X-linked ichthyosis,XLI)、先天性非大皰性魚鱗病樣紅皮病(nonbullous congenital ichthyosiform erythroderma, NCIE)。表皮松解性角化過度型魚鱗病(EHK)是一種罕見的常染色體顯性遺傳性皮膚病(MIM 113800),其發(fā)生率在0.33／10萬～1／10萬之間。約有半數(shù)病人有陽性家族史,90%的患者在1歲時(shí)開始有皮膚損害,71%出生時(shí)就有。本病具有很高的自發(fā)突變率,至少半數(shù)病例為散發(fā)性。遺傳學(xué)的連鎖分析、基因突變和轉(zhuǎn)基因小鼠研究,都表明EHK是由編碼基底層以上的角蛋白基因1(KRT1)和10(KRT10)突變引起的。本研究利用直接測序的方法對該家系進(jìn)行KRT1和KRT10基因的所有編碼區(qū),特別是熱點(diǎn)區(qū)基因突變的檢測,以期發(fā)現(xiàn)新的致病突變,進(jìn)一步豐富基因的突變譜。 材料和方法 一、實(shí)驗(yàn)材料 (一)研究對象： 經(jīng)中國醫(yī)科大學(xué)附屬第一醫(yī)院皮膚科確診的遼寧漢族表皮松解性角化過度型魚鱗病一家系,其中二人發(fā)病,為母子關(guān)系,且其臨床表現(xiàn)相同。 (二)主要試劑： 10%SDS、EDTA、硼酸、蛋白酶K、Tris飽和酚、氯仿、無水乙醇、溴化乙啶、瓊脂糖、TBE緩沖液、2×Taq plus PCR Master Mix。 二、實(shí)驗(yàn)方法 取患者皮損進(jìn)行組織病理檢查,提取該家系成員的外周血DNA,采用聚合酶連反應(yīng)(PCR)及DNA直接測序方法,檢測患者角蛋1(KRT1)及角蛋白10(KRT10)的基因突變。確診之日起予先證者第三代維甲酸(阿羅神)口服,同時(shí)配合外用藥治療。 結(jié)果 該家系2例患者存在KRT10基因的雜合點(diǎn)突變,即在KRT10基因第2140位G→A,導(dǎo)致其第156位的精氨酸變?yōu)榻M氨酸(R156H)。維甲酸治療四個(gè)月后,癥狀改善,皮膚鱗屑明顯變薄。 結(jié)論 KRT10R156H是導(dǎo)致該家系2例患者臨床表型的特異突變,進(jìn)一步證實(shí)KRT10基因第156位密碼子是突變熱點(diǎn),從而為基因診斷和基因治療提供一定的依據(jù)。維甲酸治療效果顯著。
[Abstract]:Preface ichthyosis (ichthyosis) is a group of keratosis hereditary skin diseases characterized by dry skin and lamellar scales. According to the genetic mechanism, clinical manifestations and histopathological differences, there are mainly four types: (ichthyosis vulgaris, IV), 's epidermolytic hyperkeratosis, (epidermolytic hyperkeratosis, EHK), X linked ichthyosis (X-linked ichthyosis,XLI). Congenital non-bullous ichthyosis (nonbullous congenital ichthyosiform erythroderma, NCIE). Epidermolytic hyperkeratosis (EHK) is a rare autosomal dominant hereditary dermatosis (MIM 113800) with an incidence of 0.33% ~ 1 / 100 000. About half of the patients had a positive family history, 90 per cent had skin damage at age 1 and 71 per cent were born with skin damage. The disease has a high spontaneous mutation rate, at least half of the cases are sporadic. Genetic linkage analysis, gene mutation and transgenic mouse studies showed that EHK was caused by keratin 1 (KRT1) and 10 (KRT10) mutations above the basal layer. In this study, direct sequencing was used to detect all the coding regions of KRT1 and KRT10 genes, especially the mutations in hot spots, in order to find new pathogenic mutations and further enrich the mutation profiles of the genes. Materials and methods 1. Experimental materials (1) subjects: a family of epidermolysis hyperkeratosis in Liaoning Han nationality was confirmed by the dermatology department of the first affiliated Hospital of China Medical University. Two of them had a maternal-child relationship, and their clinical manifestations were the same. (II) main reagents: SDS-EDTA, boric acid, proteinase KTIs saturated phenol, chloroform, anhydrous ethanol, ethidium bromide, agarose, TBE buffer, 2 脳 Taq plus PCR Master Mix. Secondly, the skin lesions of the patients were taken for histopathological examination. The peripheral blood DNA, of the family members was extracted by polymerase chain reaction (PCR) and DNA direct sequencing. The gene mutations of keratin 1 (KRT1) and keratin 10 (KRT10) were detected. From the date of diagnosis, the third generation of retinoic acid (aroshen) was given orally and treated with foreign medicine. Results the heterozygous point mutation of KRT10 gene was found in two patients of this family, that is, the 2140 G of KRT10 gene resulted in the transformation of the 156th arginine into histidine (R156H). After four months of retinoic acid treatment, symptoms improved and skin scales thinned significantly. Conclusion KRT10R156H is a specific mutation leading to the clinical phenotype of 2 patients in this pedigree. It is further confirmed that codon 156 of KRT10 gene is a hot spot of mutation, which provides a basis for gene diagnosis and gene therapy. The therapeutic effect of retinoic acid is remarkable.
【學(xué)位授予單位】：中國醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2010
【分類號】：R758.52
[Abstract]:Preface ichthyosis (ichthyosis) is a group of keratosis hereditary skin diseases characterized by dry skin and lamellar scales. According to the genetic mechanism, clinical manifestations and histopathological differences, there are mainly four types: (ichthyosis vulgaris, IV), 's epidermolytic hyperkeratosis, (epidermolytic hyperkeratosis, EHK), X linked ichthyosis (X-linked ichthyosis,XLI). Congenital non-bullous ichthyosis (nonbullous congenital ichthyosiform erythroderma, NCIE). Epidermolytic hyperkeratosis (EHK) is a rare autosomal dominant hereditary dermatosis (MIM 113800) with an incidence of 0.33% ~ 1 / 100 000. About half of the patients had a positive family history, 90 per cent had skin damage at age 1 and 71 per cent were born with skin damage. The disease has a high spontaneous mutation rate, at least half of the cases are sporadic. Genetic linkage analysis, gene mutation and transgenic mouse studies showed that EHK was caused by keratin 1 (KRT1) and 10 (KRT10) mutations above the basal layer. In this study, direct sequencing was used to detect all the coding regions of KRT1 and KRT10 genes, especially the mutations in hot spots, in order to find new pathogenic mutations and further enrich the mutation profiles of the genes. Materials and methods 1. Experimental materials (1) subjects: a family of epidermolysis hyperkeratosis in Liaoning Han nationality was confirmed by the dermatology department of the first affiliated Hospital of China Medical University. Two of them had a maternal-child relationship, and their clinical manifestations were the same. (II) main reagents: SDS-EDTA, boric acid, proteinase KTIs saturated phenol, chloroform, anhydrous ethanol, ethidium bromide, agarose, TBE buffer, 2 脳 Taq plus PCR Master Mix. Secondly, the skin lesions of the patients were taken for histopathological examination. The peripheral blood DNA, of the family members was extracted by polymerase chain reaction (PCR) and DNA direct sequencing. The gene mutations of keratin 1 (KRT1) and keratin 10 (KRT10) were detected. From the date of diagnosis, the third generation of retinoic acid (aroshen) was given orally and treated with foreign medicine. Results the heterozygous point mutation of KRT10 gene was found in two patients of this family, that is, the 2140 G of KRT10 gene resulted in the transformation of the 156th arginine into histidine (R156H). After four months of retinoic acid treatment, symptoms improved and skin scales thinned significantly. Conclusion KRT10R156H is a specific mutation leading to the clinical phenotype of 2 patients in this pedigree. It is further confirmed that codon 156 of KRT10 gene is a hot spot of mutation, which provides a basis for gene diagnosis and gene therapy. The therapeutic effect of retinoic acid is remarkable.
【學(xué)位授予單位】：中國醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2010
【分類號】：R758.52
【相似文獻(xiàn)】
相關(guān)期刊論文 前10條
1 本刊通訊員;;魚鱗病是可以治愈的[J];食品與健康;2001年08期
2 周升平;;部長·農(nóng)家女·皮膚頑疾[J];東北之窗;2011年16期
3 沈菲;梅小斌;;魚鱗病伴發(fā)的腎臟病[J];藥學(xué)服務(wù)與研究;2011年03期
4 ;遺傳性皮膚病[J];中國醫(yī)學(xué)文摘(皮膚科學(xué));2011年04期
5 ;名醫(yī)特別門診[J];婦女生活;2011年08期
6 蔡智波;陳恩德;;招飛體檢皮膚科淘汰統(tǒng)計(jì)及標(biāo)準(zhǔn)掌握探討[J];航空軍醫(yī);2010年04期
7 李常興;李雪梅;張錫寶;韓春雷;馬澤|，
本文編號：2359168
本文鏈接：http://sikaile.net/yixuelunwen/pifb/2359168.html
最近更新
教材專著
