骨形成蛋白4介导的酸和甘氨胆酸诱导Barrett食管的作用研究

Study on the role of bone morphogenetic protein 4 in mediating acid and glycocholic acid induced Barrett＇s esophagus

目的探讨骨形成蛋白4（BMP4）在Barrett食管发生过程中的作用机制。方法培养人食管上皮细胞（HEEC）和人成纤维细胞MRC-5。采用实时荧光定量PCR法检测不同浓度BMP4，以及不同pH值的盐酸和甘氨胆酸对HEEC中尾侧型同源转录因子2（CDX2）表达的影响，不同pH值的盐酸和甘氨胆酸对MRC-5中BMP4表达的影响。统计学分析采用t检验。结果HEEC在0.1、1.0、10.0和100.0 ng/mL的BMP4作用下，CDX2的相对表达量分别为1.617±0.246、2.489±0.455、5.629±0.449和13.670±1.689，分别高于对照组的1.000±0.043、1.029±0.094、1.001±0.002和1.049±0.051，差异均有统计学意义（t=2.47、3.14、10.31、7.47，P均〈0.05）。MRC-5在pH值为4和5的酸，pH值为4和5的甘氨胆酸的作用下，BMP4相对表达量分别为2.430±0.105、2.394±0.145、125.900±12.620、2.128±0.215，分别高于对照组的1.025±0.095、0.999±0.007、1.060±0.138、0.893±0.110，差异均有统计学意义（t=9.94、9.59、9.89、5.11，P均〈0.01）。结论BMP4能上调HEEC中CDX2的表达，从而促进Barrett食管的发生。

ObjectiveTo investigate the mechanism of bone morphogenetic protein 4（BMP4） in the genesis of Barrett′s esophagus. MethodsHuman esophageal epithelial cell （HEEC） and MRC-5 were cultured. The effects of different concentration of BMP4 and different pH value of hydrochloric acid or glycocholic acid on the expression of caudal-related homeobox transcription factor 2（CDX2） in HEEC were detected by real time polymerase chain reaction. The effects of different pH value of hydrochloric acid or glycocholic acid on BMP4 expression in MRC-5 were also investigated. Independent sample t test was performed for statistical analysis.ResultsAfter HEEC stimulated by BMP4 at 0.1, 1.0, 10.0 and 100.0 ng/mL, the relative quantity expressions of CDX2 were 1.617±0.246, 2.489±0.455, 5.629±0.449 and 13.670±1.689, respectively, which were higher than those of control group （1.000±0.043, 1.029±0.094, 1.001±0.002 and 1.049±0.051, respectively）, and the differences were statistically significant （t=2.47, 3.14, 10.31, 7.47; all P〈0.05）. After MRC-5 stimulated by acid at pH four or five, or glycocholic acid at pH four or five, the relative quantity expressions of BMP4 were 2.430±0.105, 2.394±0.145, 125.900±12.620 and 2.128±0.215, respectively, which were higher than those of control group（1.025±0.095, 0.999±0.007, 1.060±0.138 and 0.893±0.110, respectively）, and the differences were statistically significant （t=9.94, 9.59, 9.89, 5.11; all P〈0.01）.ConclusionBMP4 can increase the expression of CDX2 in HEEC, which promote the genesis of Barrett′s esophagus.