丁酸盐激活CaMKK促进紧密连接重组装的研究

Butyrate facilitates tight junction reassembly via activation of Ca^(2+)/ calmodulin-dependent protein kinase

目的探讨丁酸盐激活AMPK从而调节紧密连接的重组装的途径。方法使用丁酸盐、Ca MKK特异抑制剂(STO-609)处理上皮屏障模型Caco-2细胞,在转钙过程中检测电阻(TER)值、细胞内ATP水平以及Ca MKK和AMPK的磷酸化状况,从而判断丁酸盐促进紧密连接重组装的上游信号通路。结果丁酸盐处理Caco-2细胞后,其TER值显著增高(P<0.05),Ca MKK和AMPK均被磷酸化激活,而上述作用可被STO-609部分减弱。结论丁酸盐可通过钙离子途径磷酸化激活Ca MKK,继而活化AMPK促进紧密连接的重组。

Objective To investigate the effect of butyrate on tight junction reassembly and related pathway. Methods The Caco-2 cell monolayer model was treated with butyrate and Ca M KK inhibitor STO-609; then the transepithelial electrical resistance（ TER）,cellular ATP level,phosphorylated AM PK and Ca M KK levels were measured. Results With the treatment of butyrate,the TER of Caco-2cells significantly increased（ P〈0. 05）,phosphorylation of Ca M KK and AM PK was increased; and those effects were partly abolished by STO-609. Conclusion Butyrate may regulate the activation of AM PK through phosphorylation of Ca M KK,thus facilitate the reassembly of tight junction.