胆道闭锁致病基因的检测及功能研究
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摘要
胆道闭锁(biliary atresia, BA)是儿童最常见的严重肝脏疾病,病因至今未明,既往的研究多局限于某些炎症细胞或因子在发病中的作用,但是不同的细胞与细胞间,分子与分子间存在复杂的网络联系,这些细胞和因子哪些处于始动环节,哪些处于核心通路或是相应的下游环节,始终未能阐明。为进一步阐明该病的发病机制,本研究借助基因芯片及生物信息学技术在全基因组水平对基因表达谱及基因间的相互关系进行研究。对于所得到的关键基因对其在胆道闭锁肝脏中的表达及突变情况进行了验证。同时在体外细胞培养中对可能存在的关键调控基因的功能进行深入研究。
目的利用基因芯片技术及生物信息学方法对胆道闭锁的基因表达谱进行研究,以进一步阐明该病的发病机制。
方法Trizol一步法提取胆道闭锁组及胆总管囊肿组肝脏组织的总RNA,利用人全基因组基因表达芯片研究胆道闭锁及胆总管囊肿肝脏组织基因表达谱间的差异,并对所得差异基因进行表达趋势显著性及功能分析、信号传导通路显著性分析,并构建基因间相互作用网络,寻找在胆道闭锁发病过程中的显著性的基因表达趋势、信号通路及关键调控基因。
结果胆道闭锁与胆总管囊肿相比存在1000余条差异基因；通过对表达谱芯片所得差异基因的分析,得到：
(1)趋势21及23两个最具显著性的表达趋势。趋势21的基因功能主要涉及组织相容性抗原复合物Ⅱ介导的外源性抗原呈递作用以及由此导致的免疫反应。趋势23的基因功能主要涉及器官组织发育、细胞外基质形成,阴离子转运与磷酸基团转运。(2)趋势21涉及的显著性通路主要为细胞凋亡调控(与炎症及组织重构有关),细胞基质代谢以及细胞粘附、花生四烯酸类物质的合成(与炎症反应相关)；趋势23主要涉及细胞基质代谢(参与组织重构)、炎症反应、花生四烯酸类物质的合成、增殖与迁移能力。
(3)其中趋势21的关键基因包括ITGAM、ITGB2、TNFSFR21、RGS19,趋势23的关键基因包括LAMC1、LAMA5、MMP7、TIMP2、MMP11。
(4)通过基因调控网络构建发现LDOC1基因可能在胆道闭锁发病中起关键调控作用。
结论
(1)胆道闭锁的发病与MHCⅡ类分子介导免疫炎症及组织重构密切相关。
(2)其中ITGAM、ITGB2、TNFSFR21、RGS19和LAMC1、LAMA5、MMP7、TIMP2、MMP11分别在趋势21和趋势23中具有重要作用。
(3)LDOC1基因可能在胆道闭锁发病过程中起着核心调控作用。
目的：研究第一部分实验中发现的胆道闭锁中关键基因的表达及突变情况,以验证基因芯片结果并进一步揭示该病的发病机制。
材料与方法：
(1)22例胆道闭锁患儿和14例胆总管囊肿患儿手术活检肝脏标本,利用RT-PCR方法对趋势21和趋势23所发现的关键基因ITGAM、ITGB2、TNFRSF21、RSG19和LAMC1、LAMA5、MMP7、TIMP2、MMP11进行了半定量检测。同时以Real-time PCR的方法对LDOC1基因进行了定量检测。
(2)抽取10例胆道闭锁患儿外周静脉血2ML,抽提基因组DNA,对目的基因ITGB2和LDOC1基因外显子进行扩增纯化并测序,了解其突变及SNP改变情况。
结果：
(1)胆道闭锁肝脏组织中ITGAM、ITGB2、TNFRSF21、RSG19、LAMC1、LAMA5、MMP7、TIMP2、MMP11及LDOC1均较胆总管囊肿明显升高(P<0.05)。
(2)对ITGB2基因16个外显子进行PCR扩增、纯化、测序,测序结果与NCBI参考序列进行比对,发现：9例患儿(9/10)存在ITGB2基因单核苷酸多态性改变。这些改变包括3个突变和11个SNP改变。其中突变分别是N212Y(2/10),Y544H(1/10),P752S(4/10)；SNP改变包括：-156T/C(5/10),117G/A(1/10),919G/A(5/10),1101C/A(4/10),1533C/T(1/10),3’-UTR+123C/T(4/10),3’-UTR+140G/A(4/10),3’-UTR+148C/A(6/10),3’-UTR+170C/T(8/10),3’-UTR+219T/C(1/10),3’-UTR+286C/T(3/10)。LDOC1基因仅有1个外显子,与参考序列相比,在3例患儿(3/10)存在单核苷酸多态性改变。共有1处SNP改变,为3’UTR+822C/T。
结论
(1)胆道闭锁中,ITGAM、ITGB2、TNFRSF21、RGS19、LAMC1、LAMA5、MMP7、TIMP2、MMP11、LDOC1等关键基因表达升高,这些基因的功能主要涉及免疫炎症及细胞外基质重建,说明它们可能参与胆道闭锁炎症及纤维化的进程。
(2)胆道闭锁中ITGB2基因外显子区存在多处突变及单核苷酸改变。LDOC1基因外显子区存在1处SNP改变。这些突变可能导致相关蛋白结构及功能改变,从而参与胆道闭锁的发病。而SNP改变的存在,可能与胆道闭锁的疾病易感性相关。
目的：体外研究LDOC1基因高表达对T淋巴细胞Thl类炎症因子表达的影响。
方法：体外构建pIRES2-EGFP-LDOC1质粒,将pIRES2-EGFP-LDOC1和空白质粒pIRES2-EGFP重组并包装腺病毒,感染Jurkat细胞,利用westernblot的方法研究LDOC1蛋白的表达,利用Elisa的方法研究无转染组、pAD-LDOC1-IRES2-EGFP组和PAd-IRES2-EGFP组细胞在静息状态和PMA激活状态下Thl类炎症因子TNF-α、IL-2、IFN-γ的表达情况。
结果：成功构建pIRES2-EGFP-LDOC1质粒,并利用腺病毒包装后转染Jurkat细胞。转染LDOC1组(pAD-LDOC1-IRES2-EGFP组)较阴性转染组(PAd-IRES2-EGFP组),LDOC1蛋白表达明显升高。转染LDOC1组(pAD-LDOC1-IRES2-EGFP组)较阴性转染组(PAd-IRES2-EGFP组),在静息状态下在炎症因子产生上并无明显差异,但在经PMA刺激后,LDOC1组(pAD-LDOC1-IRES2-EGFP组)较阴性转染组(PAd-IRES2-EGFP组)TNF-α、IL-2、IFN-γ的表达均明显升高,经统计学检验具有显著性差异(p值分别为0.0099、0.0043和0.0026)。
结论：在Jurkat细胞中高表达LDOC1可以促进炎症因子TNF-α、IL-2、IFN-γ表达升高。而胆道闭锁中存在LDOC1基因表达升高,提示其可能对Thl类细胞因子表达具有促进作用,从而加剧胆道闭锁中持续而炎症的免疫损伤。
Biliary atresia (BA) is a progressive obliterative process involving the extrahepatic and intrahepatic bile ducts in the newborn. It is characterised by worsening cholestasis, hepatic fibrosis, and cirrhosis, which lead to portal hypertension and a decline in hepatic synthetic function. The etiology of BA remains unknown. It has recently been proposed that the perinatal/acquired form may be caused by a biliary trophicviral infection, leading to an initial bile duct epithelial injury that triggers a persistent immune-mediated sclerosing process resulting in obstruction of extrahepatic bile ducts. However, the relations between different genes are complicated. Therefore, we undertook a large-scale gene-expression analysis to identify physiologically relevant genomic signatures of biliary atresia in the livers of affected infants. Meanwhile, bioinformatic analysis was applied to classify the different genes and find regulatory modules in the pathgenesis of BA. Furthermore reverse-transcription polymerase chain reaction (RT-PCR) was performed to confirm changes in gene expression. We also investigated the role of the most important regulatory modules by gene transfection in votro.
Objective To investigate the gene expressing profile and find regulatory modules through large scale gene expression data analysis in BA (biliary atresia).
Methods Using cDNA microarrays, we analyzed and compared gene expression profiles in the liver tissure of BA and CBD. Meanwhile, bioinformatic analysis including STC、STC-GO (Series Test Cluster of Gene Ontology)、pathway-finder and Dynamic-GeneNet construction was applied to classify the different genes and find regulatory modules in the pathgenesis of BA.
Results There are more than 1000 different genes between BA and CBD. After STC, we find two significant profile(profile21 and 23); By STC-GO (Series Test Cluster of Gene Ontology), the different genes of profile21 and 23 mainly include immune response, antigen processing and presentation of exogenous peptide antigen via MHC classⅡand multicellular organismal development and extracellular matrix synthesis, retrospectively. By pathway-finder, integrin-mediated cell adhesion、matrix metalloproteinases、inflammatory response pathway were identified as the significant pathway. ITGAM ITGB2、TNFSFR21、RGS19 and LAMC1、LAMA5、MMP7、TIMP2、MMP11 were identified as the key genes in profile21 and 23 retrospectively. By construction of dynamic-genenet, LDOC1 was found as the most important regulatory modules.
Conclusion 1.Biliary atresia was caused by immune inflammation conducted by MHC classⅡmolecules and extracellular matrix synthesis.
2. ITGAM、ITGB2、TNFSFR21、RGS19 and LAMC1、LAMA5、MMP7、TIMP2、MMP11 have important role in the pathogenesis of immune inflammation and extracellular matrix synthesis in biliary atresia, respectively.
3. LDOC1 may be the most important regulatory module in the pathogenesis of biliary atresia.
Methods The liver biopsy tissures from 22 cases of biliary atresia and 14 cases of CBD were collected. Reverse-transcription polymerase chain reaction (RT-PCR) was performed to confirm the gene expression changes of ITGAM、ITGB2、TNFSFR21、RGS19、LAMC1、LAMA5、MMP7、TIMP2、MMP11.Real-time PCR was used to investigate the expression of LDOC1.Meawhile, screening for mutations of the ITGB2 (CD18) gene and LDOC1gene were performed on DNA extracted from whole blood of 10 biliary atresia patients. Polymerase chain reaction amplification was performed, followed by bidirectional semi-automated DNA sequencing analysis.
Results The levels of the expression of ITGAM、ITGB2、TNFSFR21、RGS19、LAMC1、LAMA5、MMP7、TIMP2、MMP11and LDOC1 mRNA were obviously higher in biliary atresia liver tissues than that in CBD liver tissues(p<0.05). After DNA sequencing analysis, ITGB2 gene showed variations in 9 bilary atresia patients (9/10), genetic variations include three mutation:N212Y(2/10), Y544H(1/10), P752S(4/10) and 11 polymorphisms:-156T/C(5/10),117G/A(1/10),919G/A(5/10),1101C/A(4/10),1533C/T(1/10), 3'-UTR+123C/T(4/10),3'-UTR+140G/A(4/10),3'-UTR+148C/A(6/10), 3'-UTR+170C/T(8/10),3'-UTR+219T/C(1/10),3'-UTR+286C/T(3/10). LDOC1 gene showed variations in 3 bilary atresia patients (3/10) including 1 polymorphisms: 3'-UTR+822C/T.
Conclusion 1.The expression of ITGAM、ITGB2、TNFSFR21、RGS19、LAMC1、LAMA5、MMP7、TIMP2、MMP11and LDOC1 elevated obviously in biliary atresia. These genes may have important role in the inflammation and liver fibrosis in biliary atresia. 2.The exons of ITGB2 gene have 3 mutations and 11 SNP variations and LDOC1 gene have 1 SNP variations in biliary atresia. The mutations may cause the constructional and functional changes of the ITGB2 protein that involved in the pathogenesis of biliary atresia. The SNP variations of LDOC1 and ITGB2 can probably be linked to a genetic predisposition to biliary atresia.
Methods We constructed plasmid pIRES2-EGFP-LDOC1 carrying a full-length LDOC1 cDNA and transfected into Jurkat cell by adenovirus. We divided into three groups including empty、pAD-LDOC1-IRES2-EGFP and PAd-IRES2-EGFP. The westernblot method was used to detect the expression of LDOC1 protein between the groups. The TNF-α、IL-2、IFN-γlevel of resting and activated Jurkat cell in each group were detected by elisa.
Results After transfection, the LDOC1 protein elevated remarkably in pAD-LDOC1-IRES2-EGFP than that of PAd-IRES2-EGFP. The expression of TNF-α、IL-2、IFN-γlevel of resting Jurkat cell were of no statistically difference between pAD-LDOCl-IRES2-EGFP and PAd-IRES2-EGFP,but when Jurkat cell was activated by PMA, the expression of TNF-α、IL-2、IFN-γlevel increased more obviously in pAD-LDOCl-IRES2-EGFP than that of PAd-IRES2-EGFP(p<0.05).
Conclusion Overexpression LDOC1 could enhance the production of TNF-α、IL-2、IFN-γ, and it indicated that LDOC1 may promote the inflammation reaction in biliary atresia which aggravate the injury of biliary epithelium.
目的利用基因芯片技术及生物信息学方法对胆道闭锁的基因表达谱进行研究,以进一步阐明该病的发病机制。
方法Trizol一步法提取胆道闭锁组及胆总管囊肿组肝脏组织的总RNA,利用人全基因组基因表达芯片研究胆道闭锁及胆总管囊肿肝脏组织基因表达谱间的差异,并对所得差异基因进行表达趋势显著性及功能分析、信号传导通路显著性分析,并构建基因间相互作用网络,寻找在胆道闭锁发病过程中的显著性的基因表达趋势、信号通路及关键调控基因。
结果胆道闭锁与胆总管囊肿相比存在1000余条差异基因；通过对表达谱芯片所得差异基因的分析,得到：
(1)趋势21及23两个最具显著性的表达趋势。趋势21的基因功能主要涉及组织相容性抗原复合物Ⅱ介导的外源性抗原呈递作用以及由此导致的免疫反应。趋势23的基因功能主要涉及器官组织发育、细胞外基质形成,阴离子转运与磷酸基团转运。(2)趋势21涉及的显著性通路主要为细胞凋亡调控(与炎症及组织重构有关),细胞基质代谢以及细胞粘附、花生四烯酸类物质的合成(与炎症反应相关)；趋势23主要涉及细胞基质代谢(参与组织重构)、炎症反应、花生四烯酸类物质的合成、增殖与迁移能力。
(3)其中趋势21的关键基因包括ITGAM、ITGB2、TNFSFR21、RGS19,趋势23的关键基因包括LAMC1、LAMA5、MMP7、TIMP2、MMP11。
(4)通过基因调控网络构建发现LDOC1基因可能在胆道闭锁发病中起关键调控作用。
结论
(1)胆道闭锁的发病与MHCⅡ类分子介导免疫炎症及组织重构密切相关。
(2)其中ITGAM、ITGB2、TNFSFR21、RGS19和LAMC1、LAMA5、MMP7、TIMP2、MMP11分别在趋势21和趋势23中具有重要作用。
(3)LDOC1基因可能在胆道闭锁发病过程中起着核心调控作用。
目的：研究第一部分实验中发现的胆道闭锁中关键基因的表达及突变情况,以验证基因芯片结果并进一步揭示该病的发病机制。
材料与方法：
(1)22例胆道闭锁患儿和14例胆总管囊肿患儿手术活检肝脏标本,利用RT-PCR方法对趋势21和趋势23所发现的关键基因ITGAM、ITGB2、TNFRSF21、RSG19和LAMC1、LAMA5、MMP7、TIMP2、MMP11进行了半定量检测。同时以Real-time PCR的方法对LDOC1基因进行了定量检测。
(2)抽取10例胆道闭锁患儿外周静脉血2ML,抽提基因组DNA,对目的基因ITGB2和LDOC1基因外显子进行扩增纯化并测序,了解其突变及SNP改变情况。
结果：
(1)胆道闭锁肝脏组织中ITGAM、ITGB2、TNFRSF21、RSG19、LAMC1、LAMA5、MMP7、TIMP2、MMP11及LDOC1均较胆总管囊肿明显升高(P<0.05)。
(2)对ITGB2基因16个外显子进行PCR扩增、纯化、测序,测序结果与NCBI参考序列进行比对,发现：9例患儿(9/10)存在ITGB2基因单核苷酸多态性改变。这些改变包括3个突变和11个SNP改变。其中突变分别是N212Y(2/10),Y544H(1/10),P752S(4/10)；SNP改变包括：-156T/C(5/10),117G/A(1/10),919G/A(5/10),1101C/A(4/10),1533C/T(1/10),3’-UTR+123C/T(4/10),3’-UTR+140G/A(4/10),3’-UTR+148C/A(6/10),3’-UTR+170C/T(8/10),3’-UTR+219T/C(1/10),3’-UTR+286C/T(3/10)。LDOC1基因仅有1个外显子,与参考序列相比,在3例患儿(3/10)存在单核苷酸多态性改变。共有1处SNP改变,为3’UTR+822C/T。
结论
(1)胆道闭锁中,ITGAM、ITGB2、TNFRSF21、RGS19、LAMC1、LAMA5、MMP7、TIMP2、MMP11、LDOC1等关键基因表达升高,这些基因的功能主要涉及免疫炎症及细胞外基质重建,说明它们可能参与胆道闭锁炎症及纤维化的进程。
(2)胆道闭锁中ITGB2基因外显子区存在多处突变及单核苷酸改变。LDOC1基因外显子区存在1处SNP改变。这些突变可能导致相关蛋白结构及功能改变,从而参与胆道闭锁的发病。而SNP改变的存在,可能与胆道闭锁的疾病易感性相关。
目的：体外研究LDOC1基因高表达对T淋巴细胞Thl类炎症因子表达的影响。
方法：体外构建pIRES2-EGFP-LDOC1质粒,将pIRES2-EGFP-LDOC1和空白质粒pIRES2-EGFP重组并包装腺病毒,感染Jurkat细胞,利用westernblot的方法研究LDOC1蛋白的表达,利用Elisa的方法研究无转染组、pAD-LDOC1-IRES2-EGFP组和PAd-IRES2-EGFP组细胞在静息状态和PMA激活状态下Thl类炎症因子TNF-α、IL-2、IFN-γ的表达情况。
结果：成功构建pIRES2-EGFP-LDOC1质粒,并利用腺病毒包装后转染Jurkat细胞。转染LDOC1组(pAD-LDOC1-IRES2-EGFP组)较阴性转染组(PAd-IRES2-EGFP组),LDOC1蛋白表达明显升高。转染LDOC1组(pAD-LDOC1-IRES2-EGFP组)较阴性转染组(PAd-IRES2-EGFP组),在静息状态下在炎症因子产生上并无明显差异,但在经PMA刺激后,LDOC1组(pAD-LDOC1-IRES2-EGFP组)较阴性转染组(PAd-IRES2-EGFP组)TNF-α、IL-2、IFN-γ的表达均明显升高,经统计学检验具有显著性差异(p值分别为0.0099、0.0043和0.0026)。
结论：在Jurkat细胞中高表达LDOC1可以促进炎症因子TNF-α、IL-2、IFN-γ表达升高。而胆道闭锁中存在LDOC1基因表达升高,提示其可能对Thl类细胞因子表达具有促进作用,从而加剧胆道闭锁中持续而炎症的免疫损伤。
Biliary atresia (BA) is a progressive obliterative process involving the extrahepatic and intrahepatic bile ducts in the newborn. It is characterised by worsening cholestasis, hepatic fibrosis, and cirrhosis, which lead to portal hypertension and a decline in hepatic synthetic function. The etiology of BA remains unknown. It has recently been proposed that the perinatal/acquired form may be caused by a biliary trophicviral infection, leading to an initial bile duct epithelial injury that triggers a persistent immune-mediated sclerosing process resulting in obstruction of extrahepatic bile ducts. However, the relations between different genes are complicated. Therefore, we undertook a large-scale gene-expression analysis to identify physiologically relevant genomic signatures of biliary atresia in the livers of affected infants. Meanwhile, bioinformatic analysis was applied to classify the different genes and find regulatory modules in the pathgenesis of BA. Furthermore reverse-transcription polymerase chain reaction (RT-PCR) was performed to confirm changes in gene expression. We also investigated the role of the most important regulatory modules by gene transfection in votro.
Objective To investigate the gene expressing profile and find regulatory modules through large scale gene expression data analysis in BA (biliary atresia).
Methods Using cDNA microarrays, we analyzed and compared gene expression profiles in the liver tissure of BA and CBD. Meanwhile, bioinformatic analysis including STC、STC-GO (Series Test Cluster of Gene Ontology)、pathway-finder and Dynamic-GeneNet construction was applied to classify the different genes and find regulatory modules in the pathgenesis of BA.
Results There are more than 1000 different genes between BA and CBD. After STC, we find two significant profile(profile21 and 23); By STC-GO (Series Test Cluster of Gene Ontology), the different genes of profile21 and 23 mainly include immune response, antigen processing and presentation of exogenous peptide antigen via MHC classⅡand multicellular organismal development and extracellular matrix synthesis, retrospectively. By pathway-finder, integrin-mediated cell adhesion、matrix metalloproteinases、inflammatory response pathway were identified as the significant pathway. ITGAM ITGB2、TNFSFR21、RGS19 and LAMC1、LAMA5、MMP7、TIMP2、MMP11 were identified as the key genes in profile21 and 23 retrospectively. By construction of dynamic-genenet, LDOC1 was found as the most important regulatory modules.
Conclusion 1.Biliary atresia was caused by immune inflammation conducted by MHC classⅡmolecules and extracellular matrix synthesis.
2. ITGAM、ITGB2、TNFSFR21、RGS19 and LAMC1、LAMA5、MMP7、TIMP2、MMP11 have important role in the pathogenesis of immune inflammation and extracellular matrix synthesis in biliary atresia, respectively.
3. LDOC1 may be the most important regulatory module in the pathogenesis of biliary atresia.
Methods The liver biopsy tissures from 22 cases of biliary atresia and 14 cases of CBD were collected. Reverse-transcription polymerase chain reaction (RT-PCR) was performed to confirm the gene expression changes of ITGAM、ITGB2、TNFSFR21、RGS19、LAMC1、LAMA5、MMP7、TIMP2、MMP11.Real-time PCR was used to investigate the expression of LDOC1.Meawhile, screening for mutations of the ITGB2 (CD18) gene and LDOC1gene were performed on DNA extracted from whole blood of 10 biliary atresia patients. Polymerase chain reaction amplification was performed, followed by bidirectional semi-automated DNA sequencing analysis.
Results The levels of the expression of ITGAM、ITGB2、TNFSFR21、RGS19、LAMC1、LAMA5、MMP7、TIMP2、MMP11and LDOC1 mRNA were obviously higher in biliary atresia liver tissues than that in CBD liver tissues(p<0.05). After DNA sequencing analysis, ITGB2 gene showed variations in 9 bilary atresia patients (9/10), genetic variations include three mutation:N212Y(2/10), Y544H(1/10), P752S(4/10) and 11 polymorphisms:-156T/C(5/10),117G/A(1/10),919G/A(5/10),1101C/A(4/10),1533C/T(1/10), 3'-UTR+123C/T(4/10),3'-UTR+140G/A(4/10),3'-UTR+148C/A(6/10), 3'-UTR+170C/T(8/10),3'-UTR+219T/C(1/10),3'-UTR+286C/T(3/10). LDOC1 gene showed variations in 3 bilary atresia patients (3/10) including 1 polymorphisms: 3'-UTR+822C/T.
Conclusion 1.The expression of ITGAM、ITGB2、TNFSFR21、RGS19、LAMC1、LAMA5、MMP7、TIMP2、MMP11and LDOC1 elevated obviously in biliary atresia. These genes may have important role in the inflammation and liver fibrosis in biliary atresia. 2.The exons of ITGB2 gene have 3 mutations and 11 SNP variations and LDOC1 gene have 1 SNP variations in biliary atresia. The mutations may cause the constructional and functional changes of the ITGB2 protein that involved in the pathogenesis of biliary atresia. The SNP variations of LDOC1 and ITGB2 can probably be linked to a genetic predisposition to biliary atresia.
Methods We constructed plasmid pIRES2-EGFP-LDOC1 carrying a full-length LDOC1 cDNA and transfected into Jurkat cell by adenovirus. We divided into three groups including empty、pAD-LDOC1-IRES2-EGFP and PAd-IRES2-EGFP. The westernblot method was used to detect the expression of LDOC1 protein between the groups. The TNF-α、IL-2、IFN-γlevel of resting and activated Jurkat cell in each group were detected by elisa.
Results After transfection, the LDOC1 protein elevated remarkably in pAD-LDOC1-IRES2-EGFP than that of PAd-IRES2-EGFP. The expression of TNF-α、IL-2、IFN-γlevel of resting Jurkat cell were of no statistically difference between pAD-LDOCl-IRES2-EGFP and PAd-IRES2-EGFP,but when Jurkat cell was activated by PMA, the expression of TNF-α、IL-2、IFN-γlevel increased more obviously in pAD-LDOCl-IRES2-EGFP than that of PAd-IRES2-EGFP(p<0.05).
Conclusion Overexpression LDOC1 could enhance the production of TNF-α、IL-2、IFN-γ, and it indicated that LDOC1 may promote the inflammation reaction in biliary atresia which aggravate the injury of biliary epithelium.
引文
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