期刊文献+
共找到76篇文章
< 1 2 4 >
每页显示 20 50 100
Potential involvement of heat shock proteins in pancreaticduodenal homeobox-1-mediated effects on the genesis of gastric cancer: A 2D gel-based proteomic study 被引量:2
1
作者 Juan Ma Bei-Bei Wang +3 位作者 Xiao-Yan Ma Wei-Ping Deng Li-Shu Xu Wei-Hong Sha 《World Journal of Gastroenterology》 SCIE CAS 2018年第37期4263-4271,共9页
AIM To identify functional proteins involved in pancreaticduodenal homeobox-1(PDX1)-mediated effects on gastric carcinogenesis.METHODS A PDX1-overexpressed model was established by transfecting gastric cancer cell lin... AIM To identify functional proteins involved in pancreaticduodenal homeobox-1(PDX1)-mediated effects on gastric carcinogenesis.METHODS A PDX1-overexpressed model was established by transfecting gastric cancer cell line SGC7901 with pcDNA3.1(+)-PDX1 vector(SGC-PDX1). Transfection with empty pcDNA3.1 vector(SGC-pcDNA) served as control. Comparative protein profiles of the two groups were analyzed by two-dimensional electrophoresis basedproteomics(2 DE gel-based proteomics). The differential proteins identified by 2 DE were further validated by qRTPCR and immunoblotting. Finally, co-immunoprecipitation was used to determine any direct interactions between PDX1 and the differential proteins.RESULTS2 DE gel proteomics identified seven differential proteins in SGC-PDX1 when compared with those in SGC-pcDNA. These included four heat shock proteins(HSPs; HSP70 p1 B, HSP70 p8, HSP60, HSP27) and three other proteins(ER60, laminin receptor 1, similar to epsilon isoform of 14-3-3 protein). Immunoblotting validated the expression of the HSPs(HSP70, HSP60, HSP27). Furthermore, their expressions were lowered to 80%, 20% and 24%, respectively, in SGC-PDX1, while PDX1 exhibited a 9-fold increase, compared to SGC-pcDNA. However, qRT-PCR analysis revealed that mRNA levels of the HSP s were increased in SGC-PDX1, suggesting that the expression of the HSP s was post-translationally regulated by the PDX1 protein. Finally, co-immunoprecipitation failed to identify any direct interaction between PDX1 and HSP70 proteins.CONCLUSION This study demonstrates the potential involvement of HSPs in PDX1-mediated effects on the genesis of gastric cancer. 展开更多
关键词 pancreatic-duodenal homeobox-1 Heat shock proteins Gastric cancer Proteomics Two-dimensional ELECTROPHORESIS
下载PDF
Down-regulation of pancreatic transcription factors and incretin receptors in type 2 diabetes 被引量:9
2
作者 Hideaki Kaneto Taka-aki Matsuoka 《World Journal of Diabetes》 SCIE CAS 2013年第6期263-269,共7页
Type 2 diabetes is one of the most prevalent and serious metabolic diseases.Under diabetic conditions,chronic hyperglycemia and subsequent induction of oxidative stress deteriorate pancreaticβ-cell function,which lea... Type 2 diabetes is one of the most prevalent and serious metabolic diseases.Under diabetic conditions,chronic hyperglycemia and subsequent induction of oxidative stress deteriorate pancreaticβ-cell function,which leads to the aggravation of type 2 diabetes.Although such phenomena are well known as glucose toxicity,its molecular mechanism remains unclear.In this review article,we describe the possible molecular mechanism forβ-cell dysfunction found in type 2 diabetes,focusing on(1)oxidative stress,(2)pancreatic transcription factors(PDX-1 and MafA)and(3)incretin receptors(GLP-1 and GIP receptors).Under such conditions,nuclear expression levels of PDX-1 and MafA are decreased,which leads to suppression of insulin biosynthesis and secretion.In addition,expression levels of GLP-1 and GIP receptors are decreased,which likely contributes to the impaired incretin effects found in diabetes.Taken together,it is likely that downregulation of pancreatic transcription factors(PDX-1and MafA)and down-regulation of incretin receptors(GLP-1 and GIP receptors)explain,at least in part,the molecular mechanism forβ-cell dysfunction found in type 2 diabetes. 展开更多
关键词 pancreatic β -CELLS Oxidative stress pancreatic duodenal homeobox-1 MAFA Incretin receptor
下载PDF
Pdx-1、Ngn3联合MafA诱导干细胞分化为胰岛素分泌细胞移植治疗1型糖尿病大鼠的效果
3
作者 闫淑芳 袁慧娟 《河南医学研究》 CAS 2024年第9期1553-1557,共5页
目的通过胰十二指肠同源盒1(Pdx-1)、神经元素3(Ngn3)联合V型肌腱膜纤维肉瘤癌基因同源基因A(MafA)共转染大鼠骨髓间充质干细胞(BMSCs)诱导分化为胰岛素分泌细胞(IPCs),移植治疗1型糖尿病大鼠并观察其疗效。方法Pdx-1、Ngn3和MafA共转染... 目的通过胰十二指肠同源盒1(Pdx-1)、神经元素3(Ngn3)联合V型肌腱膜纤维肉瘤癌基因同源基因A(MafA)共转染大鼠骨髓间充质干细胞(BMSCs)诱导分化为胰岛素分泌细胞(IPCs),移植治疗1型糖尿病大鼠并观察其疗效。方法Pdx-1、Ngn3和MafA共转染BMSCs诱导为IPCs,链脲佐菌素(STZ)制备45只1型糖尿病SD大鼠模型,BMSCs组(15只)肾被膜下移植2×10^(6) BMSCs,IPCs组(15只)大鼠肾被膜下移植2×10^(6) IPCs,sham-operation组(15只)大鼠肾被膜下注入等量生理盐水,另取15只健康SD大鼠肾被膜下注入等量生理盐水作为normal组。监测各组大鼠移植第0、7、14、21、28 d空腹血糖及体重;第21天对各组大鼠均行葡萄糖耐量试验;第28天取各组大鼠肾脏组织进行免疫组化。结果BMSCs和IPCs组大鼠血糖随时间下降,移植第21天两组大鼠空腹血糖低于移植第0天,且均低于同期sham-operation组(P<0.05),移植第28天IPCs组大鼠空腹血糖低于BMSCs组(P<0.05)。糖尿病大鼠中IPCs组和BMSCs组腹腔葡萄糖耐量实验曲线下面积小于sham-operation组,且IPCs组曲线下面积最小(P<0.05)。BMSCs组和IPCs组大鼠肾脏组织可见棕色荧光的胰岛素表达,且IPCs组胰岛素荧光光密度高于BMSCs组(P<0.05)。结论Pdx-1-Ngn3联合MafA诱导BMSCs形成的IPCs移植后在糖尿病大鼠体内可降低糖尿病大鼠的空腹血糖。 展开更多
关键词 胰十二指肠同源盒1 神经元素3 V型肌腱膜纤维肉瘤癌基因同源基因A 大鼠骨髓间充质干细胞 胰岛素分泌细胞 1型糖尿病
下载PDF
Roux-en-Y gastric bypass promotes expression of PDX-1 and regeneration of β-cells in Goto-Kakizaki rats 被引量:14
4
作者 Zhen Li,Dong-Fei Li,Jing-Xing Dai,Yu Bai,Lin Yuan,Southern Medical University,Institute of Basic Medical Anatomy National Key Disciplines,Guangzhou 510515,Guangdong Province,China Hong-Ya Zhang,Lu-Xian Lv,Wen-Qiang Li,Second Affiliated Hospital,Xinxiang Medical College,Henan Province Key Laboratory of Biological Psychiatry,Xinxiang 453002,Henan Province,China Ou Sha,Department of Anatomy,Faculty of Medicine,Shenzhen University,Shenzhen 518060,Guangdong Province,China 《World Journal of Gastroenterology》 SCIE CAS CSCD 2010年第18期2244-2251,共8页
AIM:To study the effects of Roux-en-Y gastric bypass(RYGB) on the expression of pancreatic duodenal homeobox-1(PDX-1) and pancreatic β-cell regeneration/neogenesis,and their possible mechanisms in diabetics.METHODS:T... AIM:To study the effects of Roux-en-Y gastric bypass(RYGB) on the expression of pancreatic duodenal homeobox-1(PDX-1) and pancreatic β-cell regeneration/neogenesis,and their possible mechanisms in diabetics.METHODS:Three groups of randomly selected nonobese diabetic Goto-Kakizaki(GK) rats were subjected to RYGB,sham-RYGB and sham-operation(sham-op) surgery,respectively.The rats were euthanized at postoperative 1,2,4 and 12 wk.Their pancreases were resected and analyzed using reverse transcription polymerase chain reaction to detect the mRNA of PDX-1.Anti-PDX-1 immunohistochemical(IHC) staining and Western blotting were used to detect the protein of PDX-1.Double IHC staining of anti-Brdu and-insulin was performed to detect regenerated β-cells.The index of double Brdu and insulin positive cells was calculated.RESULTS:In comparison with sham-RYGB and sham-op groups,a significant increase in the expressions of PDX-1 mRNA in RYGB group was observed at all experimental time points(1 wk:0.378 ± 0.013 vs 0.120 ± 0.010,0.100 ± 0.010,F = 727.717,P < 0.001;2 wk:0.318 ± 0.013 vs 0.110 ± 0.010,0.143 ± 0.015,F = 301.509,P < 0.001;4 wk:0.172 ± 0.011 vs 0.107 ± 0.012,0.090 ± 0.010,F = 64.297,P < 0.001;12 wk:0.140 ± 0.007 vs 0.120 ± 0.010,0.097 ± 0.015,F = 16.392,P < 0.001);PDX-1 protein in RYGB group was also increased significantly(1 wk:0.61 ± 0.01 vs 0.21 ± 0.01,0.15 ± 0.01,F = 3031.127,P < 0.001;2 wk:0.55 ± 0.00 vs 0.15 ± 0.01,0.17 ± 0.01,F = 3426.455,P < 0.001;4 wk:0.39 ± 0.01 vs 0.18 ± 0.01,0.22 ± 0.01,F = 882.909,P < 0.001;12 wk:0.41 ± 0.01 vs 0.20 ± 0.01,0.18 ± 0.01,F = 515.833,P < 0.001).PDX-1 mRNA and PDX-1 protein production showed no statistical significance between the two sham groups.Many PDX-1 positive cells could be found in the pancreatic islets of the rats in RYGB group at all time points.In addition,the percentage of Brdu-insulin double staining positive cells was higher in RYGB group than in the other two groups(1 wk:0.22 ± 0.13 vs 0.03 ± 0.06,0.03 ± 0.06,P < 0.05;2 wk:0.28 ± 0.08 vs 0.00 ± 0.00,0.03 ± 0.06,P < 0.05;4 wk:0.24 ± 0.11 vs 0.07 ± 0.06,0.00 ± 0.00,P < 0.001;12 wk:0.20 ± 0.07 vs 0.03 ± 0.06,0.00 ± 0.00,P < 0.05).CONCLUSION:RYGB can increase the expression of pancreatic PDX-1 and induce the regeneration of β-cells in GK rats.The associated regeneration of islet cells may be a possible mechanism that how RYGB could improve type 2 diabetes mellitus. 展开更多
关键词 Gastric bypass Diabetes mellitus Regene-ration β-Cells Animals pancreatic duodenal homeobox-1
下载PDF
Transcription factor PDX-1 in human colorectal adenocarcinoma:A potential tumor marker? 被引量:4
5
作者 Nikiforos Ballian Francis Charles Brunicardi 《World Journal of Gastroenterology》 SCIE CAS CSCD 2008年第38期5823-5826,共4页
AIM: To examine the expression of pancreatic duodenal homeobox-1 (PDX-1) transcription factor in human colorectal cancer. METHODS: RT-PCR, Western blotting, and immuno-histochemistry were performed to determine the ex... AIM: To examine the expression of pancreatic duodenal homeobox-1 (PDX-1) transcription factor in human colorectal cancer. METHODS: RT-PCR, Western blotting, and immuno-histochemistry were performed to determine the expression pattern of transcription factor PDX-1 in primary colorectal tumor, hepatic metastasis, and benign colon tissue from a single patient. RESULTS: The highest PDX-1 transcription levels were detected in the metastasis material. Lower levels of PDX-1 were found to be present in the primary tumor, while normal colon tissue failed to express detectable levels of PDX-1. Western blot data revealed a PDX-1 expression pattern identical to that of mRNA expression. Immunohistochemistry confirmed high metastasis PDX-1 expression, lower levels in the primary tumor, and the presence of only traces of PDX-1 in normal colon tissue. CONCLUSION: These data argue for further evaluation of PDX-1 as a biomarker for colorectal cancer. 展开更多
关键词 Colorectal cancer pancreatic duodenal homeobox-1 Tumor marker Transcription factor Diagnostics
下载PDF
PDX-1 Expression in Pancreatic Ductal Cells after Partial Pancreatectomy in Adult Rats 被引量:1
6
作者 刘涛 王春友 +3 位作者 万赤丹 熊炯忻 许逸卿 周峰 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2004年第5期464-466,共3页
To investigate the protein and mRNA expression of pancreas/duodenal homeobox-1 (PDX-1), a transcription factor as a marker for pancreatic stem cells, in pancreatic ductal cells of rats after partial (90 %) pancreatect... To investigate the protein and mRNA expression of pancreas/duodenal homeobox-1 (PDX-1), a transcription factor as a marker for pancreatic stem cells, in pancreatic ductal cells of rats after partial (90 %) pancreatectomy and evaluated the significance of the PDX-1 expression. Western blot and Reverse transcriptase-polymerase chain reaction (RT-PCR) were used to detect the expression of PDX-1 protein and mRNA respectively. PDX-1 protein was only faintly detected in pancreatic ductal cells on the day 1 after partial pancreatectomy. On the day 2 and 3 after operation in operation group, a 2—3 fold increased PDX-1 protein was observed, corresponding to the characteristic 42—kD protein in Western blot. There was significant difference between operation group and sham-operation group (P<0.05). PDX-1 protein expression on the day 5 and 7 after operation had already been no difference from control group (P>0.05). RT-PCR revealed the PDX-1 mRNA expression showed no significant difference between operation group at various time points and sham-operation group (P>0.05). These results indicate that there was overexpression of PDX-1 in the cells of pancreatic epithelium during the regeneration of remnant pancreas after partial pancreatectomy in adult rats, suggesting the pancreatic stem cells in pancreatic ductal epithelial cells are involved in the regeneration of remnant pancreas and the expression of PDX-1 in ductal cells was regulated posttranscription. 展开更多
关键词 pancreas/duodenal homeobox-1 pancreatic epithelium ductal cells rats
下载PDF
LncRNA ZEB1-AS1调节miR-429/PDX-1轴对结肠癌细胞增殖、凋亡和上皮间质转化的影响
7
作者 程广兴 郑发著 +2 位作者 唐金龙 张博 刘春辉 《中国现代普通外科进展》 CAS 2023年第10期757-761,766,共6页
目的:探究长链非编码RNA E盒锌指蛋白1反转录本1(LncRNA ZEB1-AS1)调节miR-429/胰腺十二指肠同源盒基因-1(PDX-1)轴对结肠癌细胞增殖、凋亡以及上皮间质转化(EMT)的影响。方法:q RT-PCR法测定细胞LncRNA ZEB1-AS1、miR-429、PDX-1 m RN... 目的:探究长链非编码RNA E盒锌指蛋白1反转录本1(LncRNA ZEB1-AS1)调节miR-429/胰腺十二指肠同源盒基因-1(PDX-1)轴对结肠癌细胞增殖、凋亡以及上皮间质转化(EMT)的影响。方法:q RT-PCR法测定细胞LncRNA ZEB1-AS1、miR-429、PDX-1 m RNA表达水平;将SW480细胞分为对照组、si-NC组、si-LncRNA ZEB1-AS1组、si-LncRNA ZEB1-AS1+inhibitor NC组、si-LncRNA ZEB1-AS1+miR-429 inhibitor组、si-LncRNA ZEB1-AS1+miR-429 inhibitor+sh NC组、si-LncRNA ZEB1-AS1+miR-429 inhibitor+sh PDX-1组,检测SW480细胞增殖、凋亡、EMT相关蛋白、紧密连接蛋白以及PDX-1表达;双荧光素酶报告基因实验验证LncRNA ZEB1-AS1、PDX-1与miR-429关系。结果:与人正常结肠上皮细胞相比,NCM-460、HCT116、SW620、CT26、SW480中LncRNA ZEB1-AS1、PDX-1 m RNA表达升高,miR-429表达降低,其中SW480细胞升高或降低最为显著(P<0.05);与对照组相比,si-LncRNA ZEB1-AS1组SW480细胞增殖抑制率、凋亡率及E-cad、ZO-1、Claudin-1表达升高,N-cad、Vimentin表达降低(P<0.05);miR-429 inhibitor下调可逆转LncRNA ZEB1-AS1沉默对SW480细胞的作用;si-LncRNA ZEB1-AS1与miR-429 inhibitor基础上沉默PDX-1可恢复LncRNA ZEB1-AS1沉默对SW480细胞的作用;miR-429与LncRNA ZEB1-AS1以及PDX-1均有靶向关系。结论:沉默ZEB1-AS1可能通过促进miR-429表达,抑制PDX-1表达,进而抑制SW480细胞增殖与EMT,并促进细胞凋亡。 展开更多
关键词 结肠肿瘤 E盒锌指蛋白1反转录本1 miR-429 胰腺十二指肠同源盒基因-1
下载PDF
PDX1在糖尿病中的研究进展
8
作者 黄丽红 王凤 乔永超 《华夏医学》 CAS 2023年第1期178-182,共5页
胰-十二指肠同源盒1基因(PDX1)是胰腺发育和胰岛素调控的关键基因。PDX1缺陷会抑制胰腺发育,导致高血糖。本文就PDX1的结构及其在胰岛β细胞分化、胰岛素合成与分泌调控中的作用与机制进行探讨,并就其与新生儿糖尿病、成人发病型糖尿病... 胰-十二指肠同源盒1基因(PDX1)是胰腺发育和胰岛素调控的关键基因。PDX1缺陷会抑制胰腺发育,导致高血糖。本文就PDX1的结构及其在胰岛β细胞分化、胰岛素合成与分泌调控中的作用与机制进行探讨,并就其与新生儿糖尿病、成人发病型糖尿病和2型糖尿病之间的关联进行阐述,以便为糖尿病的治疗提供新思路。 展开更多
关键词 胰-十二指肠同源盒1基因 新生儿糖尿病 成人发病型糖尿病 2型糖尿病
下载PDF
Effects of supraphysiologic concentration glucose on pancreatic duodenal homeobox-1 expression and insulin secretion in rats 被引量:1
9
作者 XIAO Chang-qing DENG Hong-ming HUANG Yun 《Chinese Medical Journal》 SCIE CAS CSCD 2007年第11期1020-1023,共4页
The islet transcription factor pancreatic duodenal homeobox-1 (PDX-1, also known as insulin promoter factor-1 or IPF-1) is an orphan homeodomain protein that plays an important role in the development, proliferation... The islet transcription factor pancreatic duodenal homeobox-1 (PDX-1, also known as insulin promoter factor-1 or IPF-1) is an orphan homeodomain protein that plays an important role in the development, proliferation, differentiation and maturation of pancreatic cells. It is initially detected in the part of the dorsal and ventral primitive gut epithelium that later develops into the pancreas in embryonic period. In adults, its expression is found predominantly in the differentiated islet beta-cells. A recent study shows that PDX-1 is a major islet transcription factor which activates insulin gene transcription.1 Glucose-induced insulin biosynthesis is concerned with some motifs in insulin gene promoter, and PDX-1 activates insulin gene transcription and biosynthesis through binding to these motifs. Targeted inactivation of PDX-1 gene in the mice as well as its mutation in humans2 results in agenesis of the pancreas. 展开更多
关键词 pancreatic duodenal homeobox-1 insulin secretion islet cells GLUCOSE
原文传递
2型糖尿病大鼠胰岛β细胞PDX-1基因表达及GLP-1的干预作用 被引量:10
10
作者 吴美芬 陈晓铭 +3 位作者 武革 郑坤杰 潘海燕 方烁 《实用医学杂志》 CAS 北大核心 2014年第11期1699-1701,共3页
目的:观察胰高血糖素样肽-1(glucagon-like peptide-1,GLP-1)类似物利拉鲁肽对2型糖尿病大鼠胰腺十二指肠同源盒-1(pancreatic duodenal homeobox-1,PDX-1)基因表达水平的影响。方法:将60只雄性SD大鼠平均分为3组:正常对照组、糖尿病未... 目的:观察胰高血糖素样肽-1(glucagon-like peptide-1,GLP-1)类似物利拉鲁肽对2型糖尿病大鼠胰腺十二指肠同源盒-1(pancreatic duodenal homeobox-1,PDX-1)基因表达水平的影响。方法:将60只雄性SD大鼠平均分为3组:正常对照组、糖尿病未治疗组及GLP-1组,正常对照组给予普通饲料喂养,糖尿病未治疗组与GLP-1组给予高脂饲料喂养,于第8周末以链脲佐菌素(Streptozotocin,STZ)30 mg/(kg·bw)腹腔注射高脂饲料喂养组,成模后GLP-1组给予利拉鲁肽200μg/kg皮下注射,每日2次共持续8周。其余2组予等量的生理盐水皮下注射。以实时荧光定量逆转录-聚合酶链反应(Real-Time fluorescent quantitative reverse transcription-polymerase chain reaction,Real-Time RT-PCR)法检测大鼠胰腺胰岛细胞PDX-1基因的表达。结果:与正常对照组比较,GLP-1组及未治疗组PDX-1 mRNA的表达量明显减低(P<0.001),GLP-1治疗后2型糖尿病大鼠胰腺PDX-1 mRNA的表达量可明显增高(P<0.001)。结论:GLP-1可上调2型糖尿病大鼠胰岛β细胞PDX-1基因的表达。 展开更多
关键词 糖尿病 2型 大鼠 PDX-1 GLP-1 利拉鲁肽
下载PDF
PDX1联合NKX6.1促进骨髓间充质干细胞分化为胰岛β样细胞 被引量:12
11
作者 唐小龙 张洹 +1 位作者 朱康儿 郭敏 《中国病理生理杂志》 CAS CSCD 北大核心 2009年第8期1591-1599,共9页
目的:为提高骨髓间充质干细胞(BMSCs)向胰腺β样细胞的分化效率,以产生足够用于移植的胰岛样细胞。方法:构建含PDX1与NKX6.1双基因的重组腺病毒载体,用重组腺病毒感染并联合多种细胞因子分步诱导BMSCs。用RT-PCR、Western blotting等多... 目的:为提高骨髓间充质干细胞(BMSCs)向胰腺β样细胞的分化效率,以产生足够用于移植的胰岛样细胞。方法:构建含PDX1与NKX6.1双基因的重组腺病毒载体,用重组腺病毒感染并联合多种细胞因子分步诱导BMSCs。用RT-PCR、Western blotting等多种方法分别检测PDX1、NKX6.1、胰岛素及C-肽表达情况;观测植入鼠肾包膜下的细胞形态与胰岛素、C-肽等相应分子表达情况以及检测植入细胞对糖尿病模型大鼠的血糖水平的调节能力。结果:BMSCs经重组腺病毒pAdxsi-CMV-PDX1/CMV-NKX6.1联合相应细胞因子分步诱导,双硫腙染色细胞质呈亮红色,RT-PCR显示诱导后的细胞持续稳定表达胰岛素、葡萄糖转运蛋白2(GLUT2)等β细胞相关分子;Western blotting、免疫细胞化学与间接荧光结果亦相似。所诱导的实验组细胞经5.5和25mmol/L葡萄糖刺激后胰岛素分泌水平分别为(1240.4±109.3)mU/L和(3539.8±245.1)mU/L,并显著高于对照组的分泌量。移植实验组细胞可恢复STZ糖尿病小鼠血糖正常水平。结论:PDX1与NKX6.1联合细胞因子在体外能有效地诱导BMSCs分化为胰岛β样细胞;这种胰岛β样细胞移植能有效恢复STZ糖尿病小鼠的血糖正常水平,维持小鼠良好的生存状态,这将为治疗糖尿病带来新的希望。 展开更多
关键词 胰腺十二指肠同源框1 NK6转录因子相关1 间充质干细胞 腺病毒载体
下载PDF
Pdx1与NeuroD1联合诱导L02细胞Nkx6.1和GLUT2的表达 被引量:3
12
作者 唐小龙 郭敏 +1 位作者 张洹 朱康儿 《中国病理生理杂志》 CAS CSCD 北大核心 2009年第1期163-167,共5页
目的:应用分子生物学技术,构建胰腺十二指肠同源框蛋白1(Pdx1)和神经分化因子1(NeuroD1)转录因子真核表达质粒,检测其能否在真核细胞中有效表达并诱导人肝细胞转分化的能力。方法:以人胚胎胰腺组织mRNA为模板经RT-PCR扩增获得Pdx1和Neur... 目的:应用分子生物学技术,构建胰腺十二指肠同源框蛋白1(Pdx1)和神经分化因子1(NeuroD1)转录因子真核表达质粒,检测其能否在真核细胞中有效表达并诱导人肝细胞转分化的能力。方法:以人胚胎胰腺组织mRNA为模板经RT-PCR扩增获得Pdx1和NeuroD1基因,分别克隆到真核双表达载体pIRES质粒的多克隆位点A(MCSA)和B(MCSB)中,构建pI/Pdx1/NeuroD1真核表达质粒,并转染L02细胞。用RT-PCR、免疫组化、间接荧光法和Western blotting检测目的基因及NK转录因子相关的Nkx6.1(Nkx6.1)和葡萄糖运载体2(GLUT2)的表达情况。结果:目的基因克隆正确且在真核细胞中有效表达;并可诱导肝细胞表达胰腺β细胞功能相关的Nkx6.1和GLUT2因子。结论:pI/Pdx1/NeuroD1质粒成功构建和在人真核细胞表达,并诱导人肝细胞表达β细胞功能相关的Nkx6.1转录因子和GLUT2,提示Pdx1与NeuroD1可诱导肝细胞向胰腺内分泌细胞分化。 展开更多
关键词 胰腺十二指肠同源盒蛋白质1 神经源分化因子1 质粒 转录因子
下载PDF
Pdx1与Ngn3协同诱导LO2细胞分化为胰腺β样细胞 被引量:3
13
作者 郭敏 唐小龙 张洹 《暨南大学学报(自然科学与医学版)》 CAS CSCD 北大核心 2008年第4期351-356,共6页
目的:胰腺十二指肠同源框蛋白1(Pdx1)与神经源素3(Ngn3)联合诱导肝实质细胞向胰腺β样细胞分化。方法:以人胚胎胰腺组织基因组的mRNA为模板经RT-PCR扩增获得Pdx1与Ngn3基因。应用分子生物学技术,分别克隆到真核表达载体pEGFP-C1的多克... 目的:胰腺十二指肠同源框蛋白1(Pdx1)与神经源素3(Ngn3)联合诱导肝实质细胞向胰腺β样细胞分化。方法:以人胚胎胰腺组织基因组的mRNA为模板经RT-PCR扩增获得Pdx1与Ngn3基因。应用分子生物学技术,分别克隆到真核表达载体pEGFP-C1的多克隆位点中,构建pEGFP-C1/Pdx1与pEGFP-C1/Ngn3真核表达质粒,并共转染L02细胞,用RT-PCR、免疫组化、间接荧光法检测目的基因及胰岛素相关基因葡萄糖转运体2(GLUT2)与胰岛素的表达情况。结果:目的基因克隆正确,RT-PCR、免疫组化、间接荧光证实转染后LO2细胞中有Pdx1、Ngn3和胰岛素及胰岛素相关基因GLUT2的表达。结论:Pdx1与Ngn3协同作用可诱导肝实质细胞向胰腺β样细胞分化。 展开更多
关键词 胰腺十二指肠同源框蛋白1 神经源素3 LO2细胞 转分化 胰腺β样细胞
下载PDF
血管紧张素Ⅱ对RIN-m细胞PDX-1 mRNA及蛋白表达的影响 被引量:3
14
作者 刘敏 蔡德鸿 +5 位作者 张桦 袁小澎 孙嘉 李明 张振 陈宏 《中国糖尿病杂志》 CAS CSCD 北大核心 2008年第12期749-751,共3页
目的探讨血管紧张素Ⅱ(ATⅡ)对胰十二指肠同源盒1(PDX-1)表达的影响及其与胰岛素(Ins)分泌的关系。方法测定RIN-m细胞在基础、ATⅡ及氯沙坦作用下Ins分泌量、细胞内Ins含量及细胞内PDX-1 mRNA和蛋白的表达水平。结果在100nmol/LATⅡ作用... 目的探讨血管紧张素Ⅱ(ATⅡ)对胰十二指肠同源盒1(PDX-1)表达的影响及其与胰岛素(Ins)分泌的关系。方法测定RIN-m细胞在基础、ATⅡ及氯沙坦作用下Ins分泌量、细胞内Ins含量及细胞内PDX-1 mRNA和蛋白的表达水平。结果在100nmol/LATⅡ作用下,葡萄糖刺激后Ins分泌量、细胞内Ins含量及PDX-1表达水平均明显降低(P均<0.05)。上述作用可被ATⅡ受体阻滞剂氯沙坦部分逆转。结论抑制PDX-1表达水平可能是ATⅡ影响胰岛β细胞Ins合成及分泌的机制之一。 展开更多
关键词 血管紧张素Ⅱ RIN—m细胞 胰岛素分泌 胰十二指肠同源盒1
下载PDF
烟酰胺单核苷酸对RIN-m5f细胞中胰岛素分泌及PDX-1和FoxO1基因表达的影响(英文) 被引量:3
15
作者 盛飞凤 任贤 +7 位作者 戴幸平 徐潇静 董敏 裴奇 屈健 周智广 周宏灏 刘昭前 《中南大学学报(医学版)》 CAS CSCD 北大核心 2011年第10期958-963,共6页
目的:在细胞水平研究烟酰胺单核苷酸(nicotinamide mononucleotide,NMN)对胰岛素分泌的调节作用及其对与胰岛素分泌相关的重要转录因子胰十二指肠同源盒基因(pancreatic and duodenalhomeobox-1,PDX-1)和分叉头框家族转录因子1(forkhead... 目的:在细胞水平研究烟酰胺单核苷酸(nicotinamide mononucleotide,NMN)对胰岛素分泌的调节作用及其对与胰岛素分泌相关的重要转录因子胰十二指肠同源盒基因(pancreatic and duodenalhomeobox-1,PDX-1)和分叉头框家族转录因子1(forkhead box-containing protein O-1,FoxO1)基因表达的影响。方法:采用大鼠胰岛素ELISA试剂盒检测RIN-m5f细胞胰岛素分泌水平。用Real-time PCR检测RIN-m5f细胞PDX-1和FoxO1的mRNA表达水平。用Western印迹检测RIN-m5f细胞PDX-1蛋白表达水平。结果:用瑞格列奈10 nmol/L+NMN 100μmol/L处理RIN-m5f细胞48 h,与空白对照及DMSO对照组相比,胰岛素分泌量均显著增高(P<0.05);与NMN 50μmol/L组比较,胰岛素分泌量的增高也有统计学意义(P<0.05)。10,50和100μmol/L的NMN作用RIN-m5f细胞36 h,PDX-1的mRNA表达量均上调(依次为P<0.05,P<0.01,P<0.001)。100μmol/L剂量组与10μmol/L和50μmol/L剂量组比较差异也有统计学意义(P<0.001)。50,100和200μmol/L的NMN作用RIN-m5f细胞36或48 h,PDX-1的蛋白表达量与对照组比较差异无统计学意义(P>0.05)。结论:NMN可以调控RIN-m5f细胞中胰岛素的分泌及PDX-1的mRNA表达水平。 展开更多
关键词 烟酰胺单核苷酸 胰十二指肠同源盒基因 分叉头框家族转录因子1 RIN-m5f
下载PDF
NKX6.1协同PDX1诱导人胎肝间充质干细胞分化为胰岛B样细胞 被引量:2
16
作者 唐小龙 张荣波 汪雪峰 《第二军医大学学报》 CAS CSCD 北大核心 2010年第3期258-263,共6页
目的探讨NKX6.1对PDX1诱导人胎肝间充质干细胞(fetal liver-derived mesenchymal stem cells,FL-MSCs)向胰岛B细胞分化的协同作用及可能机制,以获取足够用于移植的胰岛B样细胞。方法构建含PDX1与NKX6.1双基因的重组腺病毒载体,用该载体... 目的探讨NKX6.1对PDX1诱导人胎肝间充质干细胞(fetal liver-derived mesenchymal stem cells,FL-MSCs)向胰岛B细胞分化的协同作用及可能机制,以获取足够用于移植的胰岛B样细胞。方法构建含PDX1与NKX6.1双基因的重组腺病毒载体,用该载体感染FL-MSCs并联合相应的细胞因子分步诱导,检测诱导后的细胞中PDX1和NKX6.1基因以及NGN3、NeuroD1/Beta2、MafA因子和胰岛素等多种胰腺B细胞相关分子的表达情况。结果腺病毒载体转染24h后FL-MSCs细胞即开始表达PDX1与NKX6.1基因,检测显示诱导后的细胞先后开始表达NGN3、NeuroD1和MafA等因子,持续稳定表达胰岛素等B细胞相关分子;且这些分子表达存在时序性。结论PDX1与NKX6.1联合细胞因子在体外能有效诱导FL-MSCs分化为胰岛B样细胞;可能是通过先后活化的NGN3、NeuroD1和MafA等转录因子,限定细胞向内分泌前体细胞、进一步向B内分泌前体细胞、B细胞分化发育。 展开更多
关键词 胰腺十二指肠同源框 NKX. 胎肝 间充质干细胞 腺病毒载体 前体细胞 转录因子 胰岛素 细胞分化 细胞因子诱导
下载PDF
pdx-1基因克隆及其在人肝癌细胞系SMMC-7721中的表达 被引量:2
17
作者 陈元晓 李文林 +5 位作者 何志颖 巴月 田明 訾晓渊 张彦 胡以平 《癌变.畸变.突变》 CAS CSCD 2006年第2期81-83,共3页
背景与目的克隆小鼠胰十二指肠同源框-12(PEGFP-C1pdx-1)基因,并构建pdx-1的表达载体。材料与方法通过RT-PCR方法从小鼠胰腺总RNA中克隆pdx-1基因,并将该基因构建到pEGFP-C1和pcDNA3.0中获得pdx-1的表达载体。通过观察荧光和RT-PCR方法... 背景与目的克隆小鼠胰十二指肠同源框-12(PEGFP-C1pdx-1)基因,并构建pdx-1的表达载体。材料与方法通过RT-PCR方法从小鼠胰腺总RNA中克隆pdx-1基因,并将该基因构建到pEGFP-C1和pcDNA3.0中获得pdx-1的表达载体。通过观察荧光和RT-PCR方法检测表达载体转染SMMC-7721后pdx-1基因在细胞中的表达。结果克隆了883bp的pdx-1全长cDNA,成功构建了pEGFP-C1pdx-1和pcDNA3pdx-1两种表达载体。pEGFP-C1pdx-1转染SMMC-7721后,可观察到绿色荧光仅局限于细胞核中;通过RT-PCR方法在转染后的细胞中都检测到了pdx-1基因的转录。结论pdx-1表达载体的构建为进一步研究pdx-1基因在胰腺发育和肝干细胞增殖与分化方面的作用奠定了基础。 展开更多
关键词 胰十二指肠同源框-1(pdx-1) 基因克隆 基因表达
下载PDF
Superfect高效介导PDX-1基因在骨髓间质干细胞表达 被引量:2
18
作者 孙吉平 贾延劼 +1 位作者 王晓莉 杨于嘉 《中国病理生理杂志》 CAS CSCD 北大核心 2007年第1期12-17,共6页
目的:构建含有胰腺十二指肠同源框1(PDX-1)基因的真核表达载体,并提高重组载体在骨髓间质干细胞(MSCs)的表达效率,为骨髓MSCs作为糖尿病基因治疗的靶细胞奠定基础。方法:RT-PCR体外扩增PDX-1基因,双酶切后整合入相同双酶切的真核表达载... 目的:构建含有胰腺十二指肠同源框1(PDX-1)基因的真核表达载体,并提高重组载体在骨髓间质干细胞(MSCs)的表达效率,为骨髓MSCs作为糖尿病基因治疗的靶细胞奠定基础。方法:RT-PCR体外扩增PDX-1基因,双酶切后整合入相同双酶切的真核表达载体构建重组载体,对重组载体进行酶切分析和序列测定进行鉴定;利用Percoll细胞分离液体外培养大鼠骨髓MSCs,流式细胞仪检测细胞周期后,Superfect介导正确重组的载体转染骨髓MSCs,检测转染效率和细胞存活率从而对转染条件进行优化;G418筛选阳性细胞后,RT-PCR和Western blotting检测PDX-1基因在骨髓MSCs中的表达。结果:重组载体双酶切分析和序列测定证实重组载体构建成功。流式细胞仪显示85.9%的MSCs处于G0/G1期,提示骨髓MSCs具有强大的分化潜能;荧光显微镜下成功转染的细胞呈现全细胞绿色荧光,Superfect介导的转染效率和细胞存活率与其绝对用量和孵育细胞的时间有关;RT-PCR显示转染后PDX-1基因在骨髓MSCs表达,随转染时间延长,表达逐渐增强,与Western blotting结果一致。结论:成功构建了含有PDX-1基因的真核表达载体;Superfect能高效介导外源性基因在骨髓MSCs中表达;转染外源性基因的MSCs可望成为一种理想的基因治疗的载体细胞。 展开更多
关键词 骨髓间质干细胞 基因 胰腺十二指肠同源框1 糖尿病
下载PDF
pEGFP-N1/PDX1真核载体的构建与表达 被引量:1
19
作者 唐小龙 郭敏 +1 位作者 张洹 朱康儿 《暨南大学学报(自然科学与医学版)》 CAS CSCD 北大核心 2008年第2期121-124,共4页
目的:构建胰腺十二指肠同源框蛋白1(pancreatic and duodenal homeobox factor1,Pdx1)转录因子真核表达质粒,并检测其在真核细胞中的表达能力及其生物活性。方法:以人胚胎胰腺组织基因组的mRNA为模板经RT-PCR扩增获得Pdx1基因编码序列,... 目的:构建胰腺十二指肠同源框蛋白1(pancreatic and duodenal homeobox factor1,Pdx1)转录因子真核表达质粒,并检测其在真核细胞中的表达能力及其生物活性。方法:以人胚胎胰腺组织基因组的mRNA为模板经RT-PCR扩增获得Pdx1基因编码序列,克隆到真核表达载体pEGFP-N1的多克隆位点中,构建pEGFP-N1/Pdx1真核表达质粒,并转染L02细胞,用RT-PCR、免疫组化、间接荧光法和Western Blot检测目的基因表达情况。结果:酶切与测序结果表明目的基因序列克隆正确;RT-PCR检测目的基因Pdx1 mRNA在靶细胞L02中表达;免疫组化和间接荧光结果证实Pdx1主要存在于细胞核内;Western blot检测到细胞核内Pdx1目的蛋白。结论:完成人Pdx1基因克隆,成功构建了目的基因Pdx1真核表达载体,并在L02细胞中获得有效表达。 展开更多
关键词 胰腺十二指肠同源框蛋白1 质粒 转录因子 内切酶
下载PDF
枸杞多糖对2型糖尿病大鼠胰腺β细胞PDX-1基因表达的实验研究 被引量:11
20
作者 刘峰 黄国栋 +1 位作者 朱凌燕 汤佳珍 《中国医药科学》 2013年第1期34-36,45,共4页
目的探讨枸杞多糖对2型糖尿病大鼠β细胞分泌功能及PDX-1基因mRNA表达的影响。方法用高糖饲喂并结合链脲佐菌素诱导制备2型糖尿病大鼠模型,分组灌胃给予不同浓度的枸杞多糖及对照药物,50只实验动物随机分为5组:对照组、模型组、模型+30m... 目的探讨枸杞多糖对2型糖尿病大鼠β细胞分泌功能及PDX-1基因mRNA表达的影响。方法用高糖饲喂并结合链脲佐菌素诱导制备2型糖尿病大鼠模型,分组灌胃给予不同浓度的枸杞多糖及对照药物,50只实验动物随机分为5组:对照组、模型组、模型+30mg/kgLBP组、模型+60mg/kgLBP组、模型+90mg/kgLBP组。连续8周后检测空腹血糖、三酰甘油、总胆固醇、高密度脂蛋白、低密度脂蛋白、血清胰岛素,处死大鼠取胰腺组织,RT-PCR检测β细胞内PDX-l及胰岛素mRNA的表达水平。结果 2型糖尿病大鼠胰腺β细胞PDX-l及胰岛素mRNA的表达水平显著下降(P<0.01);60mg/kg和90mg/kgLBP组能显著上调2型糖尿病大鼠胰腺β细胞PDX-l与胰岛素mRNA表达水平(P<0.05)。结论 LBP可以通过上调PDX-l和胰岛素mRNA表达,改善2型糖尿病大鼠β细胞胰岛素合成和分泌功能的损伤。 展开更多
关键词 枸杞多糖 2型糖尿病 Β细胞 PDX-1
下载PDF
上一页 1 2 4 下一页 到第
使用帮助 返回顶部