中华肾脏病杂志
中華腎髒病雜誌
중화신장병잡지
2012年
4期
312-317
,共6页
范秋灵%李卅立%蒲实%郭佳音%岳媛%张玉侠%冯江敏%马健飞%姜奕%王力宁
範鞦靈%李卅立%蒲實%郭佳音%嶽媛%張玉俠%馮江敏%馬健飛%薑奕%王力寧
범추령%리삽립%포실%곽가음%악원%장옥협%풍강민%마건비%강혁%왕력저
糖尿病肾病%多态性,单核苷酸%候选基因%KK/Ta小鼠%多配体蛋白聚糖4
糖尿病腎病%多態性,單覈苷痠%候選基因%KK/Ta小鼠%多配體蛋白聚糖4
당뇨병신병%다태성,단핵감산%후선기인%KK/Ta소서%다배체단백취당4
Diabetic nephropathies%Polymorphism,single nucleotide%Candidate gene%KK/Ta mice%Syndecan-4
目的 在2型糖尿病肾病(DN)白蛋白尿易感基因定位的基础上,进一步筛选白蛋白尿易感基因位点(UA-1)区域附近的候选基因.方法 提取20周龄雄性KK/Ta(n=3)和BALB/c (n=2)小鼠肾脏总RNA,应用Affymetrix Murine Genome U74Av2基因芯片检测肾脏基因表达谱.选择UA-1区域的差异表达基因多配体蛋白聚糖4(syndecan-4),竞争性RT-PCR验证基因芯片的结果.提取KK/Ta、BALB/c小鼠的基因组DNA,进行syndecan-4基因编码区和启动子区域的序列分析.结果 在2型糖尿病KK/Ta小鼠UA-1区域附近存在着约10个差异表达基因.其中syndecan-4在20周龄KK/Ta小鼠肾脏中的表达上调,为BALB/c小鼠的26.1倍.在syndecan-4基因编码区存在2个基因多态性,分别为A93C和T216C多态性,2者均为同义突变.在syndecan-4基因启动子区域存在3个基因多态性,分别为-T263C、-T396C 与-G669A多态性.TATA框位于转录起始位点上游321 bp处,-T263C处恰好为转录因子Clox 的结合位点.结论 syndecan-4基因位于2型糖尿病UA-1附近区域,在20周龄KK/Ta小鼠肾脏中的表达明显上调,是DN的候选基因.syndecan-4启动子处的基因多态性可能为其差异表达的原因.
目的 在2型糖尿病腎病(DN)白蛋白尿易感基因定位的基礎上,進一步篩選白蛋白尿易感基因位點(UA-1)區域附近的候選基因.方法 提取20週齡雄性KK/Ta(n=3)和BALB/c (n=2)小鼠腎髒總RNA,應用Affymetrix Murine Genome U74Av2基因芯片檢測腎髒基因錶達譜.選擇UA-1區域的差異錶達基因多配體蛋白聚糖4(syndecan-4),競爭性RT-PCR驗證基因芯片的結果.提取KK/Ta、BALB/c小鼠的基因組DNA,進行syndecan-4基因編碼區和啟動子區域的序列分析.結果 在2型糖尿病KK/Ta小鼠UA-1區域附近存在著約10箇差異錶達基因.其中syndecan-4在20週齡KK/Ta小鼠腎髒中的錶達上調,為BALB/c小鼠的26.1倍.在syndecan-4基因編碼區存在2箇基因多態性,分彆為A93C和T216C多態性,2者均為同義突變.在syndecan-4基因啟動子區域存在3箇基因多態性,分彆為-T263C、-T396C 與-G669A多態性.TATA框位于轉錄起始位點上遊321 bp處,-T263C處恰好為轉錄因子Clox 的結閤位點.結論 syndecan-4基因位于2型糖尿病UA-1附近區域,在20週齡KK/Ta小鼠腎髒中的錶達明顯上調,是DN的候選基因.syndecan-4啟動子處的基因多態性可能為其差異錶達的原因.
목적 재2형당뇨병신병(DN)백단백뇨역감기인정위적기출상,진일보사선백단백뇨역감기인위점(UA-1)구역부근적후선기인.방법 제취20주령웅성KK/Ta(n=3)화BALB/c (n=2)소서신장총RNA,응용Affymetrix Murine Genome U74Av2기인심편검측신장기인표체보.선택UA-1구역적차이표체기인다배체단백취당4(syndecan-4),경쟁성RT-PCR험증기인심편적결과.제취KK/Ta、BALB/c소서적기인조DNA,진행syndecan-4기인편마구화계동자구역적서렬분석.결과 재2형당뇨병KK/Ta소서UA-1구역부근존재착약10개차이표체기인.기중syndecan-4재20주령KK/Ta소서신장중적표체상조,위BALB/c소서적26.1배.재syndecan-4기인편마구존재2개기인다태성,분별위A93C화T216C다태성,2자균위동의돌변.재syndecan-4기인계동자구역존재3개기인다태성,분별위-T263C、-T396C 여-G669A다태성.TATA광위우전록기시위점상유321 bp처,-T263C처흡호위전록인자Clox 적결합위점.결론 syndecan-4기인위우2형당뇨병UA-1부근구역,재20주령KK/Ta소서신장중적표체명현상조,시DN적후선기인.syndecan-4계동자처적기인다태성가능위기차이표체적원인.
Objective To identify the candidate genes in the vicinity of a susceptibility locus (urinary albumin 1,UA-1) contributing to the development of albuminuria in type 2 diabetic KK/Ta mice. Methods Total RNA was extracted from the kidneys of KK/Ta (n=3) and BALB/c (n=2) mice at 20 weeks of age.The gene expression profile in kidney was investigated using the Affymetrix Murine Genome U74Av2 array.Competitive RT-PCR was used to confirm the differential expression of syndecan-4 which located in the vicinity of UA-1.Genome DNA was extracted from KK/Ta and BALB/c mice.DNA sequence analysis of the coding and promotor region of syndecan-4 gene was conducted. Results In the vicinity of the susceptibility locus (UA-1)contributing to the development of albuminuria in type 2 diabetic KK/Ta mice,10 candidate genes that showed differential expression were identified.Among them,the gene expression of syndecan-4in KK/Ta kidneys at 20 weeks of age was up-regulated by 26.1 times of age-matched BALB/c kidneys.Sequence analysis revealed two synonymous polymorphisms in the coding region (A93C and T216C) and three polymorphisms in the promoter region (-T263C,-T396C and -G669A) of the syndecan-4 gene.The TATA box was found at 321 bp upstream from the transcription start site,and the T263C polymorphism was located in the binding site of transcription factor Clox.Conclusions Syndecan-4 gene is mapped in the vicinity of the susceptibility locus contributing to the development of albuminuria in type 2 diabetes.The gene expression of syndecan-4 in KK/Ta kidneys is up-regulated than that in age-matched BALB/c kidneys at 20 weeks of age.Thus syndecan-4 may be one of the potential candidate genes responsible for diabetic nephropathy.Sequence differences in the promoter region influence the expression levels of syndecan-4 genes in KK/Ta kidneys.
