SLC26A3与NHERF4在溃疡性结肠炎小鼠肠道组织中的动态表达研究
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摘要
目的观察分析钠氢转运体调节因子4(NHERF4)和溶质载体26A3(SLC26A3)在小鼠结肠炎模型炎症发展过程中的动态表达,为后期研究溃疡性结肠炎(UC)发病机制提供实验基础。方法以C57BL/6J小鼠为研究对象,采用恶唑酮(OXZ)诱导法制造UC模型,分别在第6、8、10、14天处死小鼠,运用疾病活动指数(DAI)评价小鼠结肠炎严重程度;采用实时荧光定量PCR(qRT-PCR)和Western blot法分别检测小鼠肠道中SLC26A3及NHERF4的mRNA和蛋白水平变化。结果与同期对照组相比,实验组第8天小鼠肠黏膜中SLC26A3的mRNA量降低(P <0. 05),而NHERF4的mRNA量增高(P <0. 05),结肠炎活动恢复过程中SLC26A3的蛋白表达呈先降低后增高的趋势,与NHERF4的蛋白表达及DAI评分呈负相关性,NHERF4的蛋白表达呈先增高后降低的趋势,与DAI评分变化趋势同步。结论在小鼠结肠炎的进展过程中,SLC26A3变化趋势与NHERF4的变化趋势相反,提示在UC发生发展过程中NHERF4可能参与调节SLC26A3的表达改变,但该结论需要进一步的干预实验进行验证。
Objective To investigate and analyze the expression of sodium/proton exchanger regulatory factor 4( NHERF4) and solute carrier 26A3( SLC26A3) in colonic mucosa of mice with oxazolone( OXZ)-induced colitis for providing an experimental date to comprehend the pathogenesis of ulcerative colitis( UC). Methods C57BL/6J mice were used as experimental animals,and model of ulcerative colitis was prepared by OXZ. Mice were sacrificed at 6 d,8 d,10 d and 14 d respectively. The disease activity index( DAI) was applied to evaluate the severity of intestinal disease in mice. qRT-PCR and Western blot were conducted to detect the mRNA and protein content of SLC26A3 and NHERF4 respectively. Results Compared with the control group,the mRNA of SLC26A3 in the mice with colitis decreased at the 8th day( P < 0. 05),while the mRNA of NHERF4 increased( P < 0. 05). It was found that the expression tendency of SLC26A3 protein was negatively correlated with the tendency of NHERF4 expression and DAI value,while the expression tendency of NHERF4 was similar with that of DAI value. Conclusion The expression tendency of SLC26A3 is opposite to that of NHERF4 during the progression of colitis in mice,suggesting that NHERF4 may be involved in the regulation of the expression of SLC26A3 during the development of UC,but need to be verified by intervene experiments.
Objective To investigate and analyze the expression of sodium/proton exchanger regulatory factor 4( NHERF4) and solute carrier 26A3( SLC26A3) in colonic mucosa of mice with oxazolone( OXZ)-induced colitis for providing an experimental date to comprehend the pathogenesis of ulcerative colitis( UC). Methods C57BL/6J mice were used as experimental animals,and model of ulcerative colitis was prepared by OXZ. Mice were sacrificed at 6 d,8 d,10 d and 14 d respectively. The disease activity index( DAI) was applied to evaluate the severity of intestinal disease in mice. qRT-PCR and Western blot were conducted to detect the mRNA and protein content of SLC26A3 and NHERF4 respectively. Results Compared with the control group,the mRNA of SLC26A3 in the mice with colitis decreased at the 8th day( P < 0. 05),while the mRNA of NHERF4 increased( P < 0. 05). It was found that the expression tendency of SLC26A3 protein was negatively correlated with the tendency of NHERF4 expression and DAI value,while the expression tendency of NHERF4 was similar with that of DAI value. Conclusion The expression tendency of SLC26A3 is opposite to that of NHERF4 during the progression of colitis in mice,suggesting that NHERF4 may be involved in the regulation of the expression of SLC26A3 during the development of UC,but need to be verified by intervene experiments.
引文
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[11]石涛,卫江鹏,刘刚,等.溃疡性结肠炎患者的肠屏障功能观察[J].中华医学杂志,2015,95(24):1941-3.
[12]Anbazhagan A N,Priyamvada S,Alakkam A,et al.Transcriptional modulation of SLC26A3(DRA)by sphingosine-1-phosphate[J].Am J Physiol Gastrointest Liver Physiol,2016,310(11):G1028-35.
[13]Ding X,Li D,Li M,et al.SLC26A3(DRA)prevents TNF-alpha-induced barrier ysfunction and dextran sulfate sodium-induced acute colitis[J].Lab Invest,2018,98(4):462-76.
[14]Chang J,Leong R W,Wasinger V C,et al.Impaired intestinal permeability contributes to ongoing bowel symptoms in patients with inflammatory bowel disease and mucosal healing[J].Gastroenterology,2017,153(3):723-31.
[15]Reid H M,Turner E C,Mulvaney E P,et al.Interaction of the human prostacyclin receptor and the NHERF4 family member intestinal and kidney enriched PDZ protein(IKEPP)[J].Biochim Biophys Acta,2012,1823(10):1998-2012.
[2]Ungaro R,Mehandru S,Allen P B,et al.Ulcerative colitis[J].Lancet,2017,389(10080):1756-70.
[3]邵晓晓,闵小彦,夏宣平,等.溶质相关载体26A3基因多态性与溃疡性结肠炎的相关性[J].中华医学遗传学杂志,2017,34(2):255-60.
[4]Xiao F,Yu Q,Li J,et al.Slc26a3 deficiency is associated with loss of colonic HCO3-secretion,absence of a firm mucus layer and barrier impairment in mice[J].Acta Physiol,2014,211(1):161-75.
[5]Xu L,Xiao F,He J,et al.Lysophosphatidic acid increases SLC26A3 expression in inflamed intestine and reduces diarrheal severity in C57BL/6 mice with dextran-sodium-sulfate-induced colitis[J].Chin Med J(Engl),2014,127(9):1737-43.
[6]史小翠,冯小燕,宋然,等.帕金森大鼠消化道组织中蛋白质NHERF家族和CAL的表达变化[J].首都医科大学学报,2013,34(2):232-8.
[7]Lee J H,Nam J H,Park J,et al.Regulation of SLC26A3 activity by NHERF4 PDZ-mediated interaction[J].Cell Signal,2012,24(9):1821-30.
[8]Sutherland L R,Martin F.5-Aminosalicylic acid enemas in treatment of distal ulcerative colitis and proctitis in Canada[J].Dig Dise Sci,1987,32(12):64s-6s.
[9]苏乐乐,张吉翔,董卫国.肠外营养治疗的109例溃疡性结肠炎患者低钙血症发生率和相关因素分析[J].中华消化杂志,2018,38(1):53-6.
[10]刘晓昌,梅俏,许建明,等.溃疡性结肠炎患者肠黏膜通透性的改变[J].安徽医科大学学报,2010,45(4):545-8.
[11]石涛,卫江鹏,刘刚,等.溃疡性结肠炎患者的肠屏障功能观察[J].中华医学杂志,2015,95(24):1941-3.
[12]Anbazhagan A N,Priyamvada S,Alakkam A,et al.Transcriptional modulation of SLC26A3(DRA)by sphingosine-1-phosphate[J].Am J Physiol Gastrointest Liver Physiol,2016,310(11):G1028-35.
[13]Ding X,Li D,Li M,et al.SLC26A3(DRA)prevents TNF-alpha-induced barrier ysfunction and dextran sulfate sodium-induced acute colitis[J].Lab Invest,2018,98(4):462-76.
[14]Chang J,Leong R W,Wasinger V C,et al.Impaired intestinal permeability contributes to ongoing bowel symptoms in patients with inflammatory bowel disease and mucosal healing[J].Gastroenterology,2017,153(3):723-31.
[15]Reid H M,Turner E C,Mulvaney E P,et al.Interaction of the human prostacyclin receptor and the NHERF4 family member intestinal and kidney enriched PDZ protein(IKEPP)[J].Biochim Biophys Acta,2012,1823(10):1998-2012.