MAPK信号通路在复方丹参注射液调节人羊膜上皮细胞AQP3表达中的作用研究

The Role of MAPK Signal Pathway in the Regulation of AQP3 Expression Induced by Compound Danshen Injection in Human Amniotic Epithelial Cells

目的探讨丝裂原活化蛋白激酶(mitogen-activated protein kinases,MAPKs)―细胞外信号调节激酶1和2(extracellular signal regulated kinase1/2,ERK1/2)信号转导通路在复方丹参注射液对人足月妊娠羊膜上皮细胞水通道蛋白3(aquaporin3,AQP3)表达调节中的作用。方法原代培养足月妊娠羊水量正常与羊水过少人羊膜上皮细胞,将人羊膜上皮细胞均分为4组:分别为空白对照组、U0126组、复方丹参注射液组和U0126加复方丹参注射液组。采用免疫印迹技术检测人羊膜上皮细胞中磷酸化ERK1/2(p-ERK1/2)和AQP3的表达。结果 (1)在羊水量正常和羊水过少人羊膜上皮细胞中,均发现p-ERK1/2表达水平在U0126组较空白对照组明显下降(P<0.05),而复方丹参注射液组可以明显上调p-ERK1/2表达水平(P<0.05),U0126加复方丹参注射液组人羊膜上皮细胞中p-ERK1/2表达水平高于U0126组,但低于复方丹参注射液组(P<0.05)。(2)在羊水量正常的人羊膜上皮细胞中,U0126组AQP3的表达与空白对照组比较无明显变化(P>0.05);复方丹参注射液组和U0126加复方丹参注射液组AQP3的表达水平均高于空白对照组(P<0.05),但两组间比较差异无统计学意义(P>0.05)。(3)在羊水过少羊膜上皮细胞中,U0126组AQP3的表达较空白对照组下降(P<0.05);复方丹参注射液组可以明显上调AQP3的表达水平(P<0.05),而U0126加复方丹参注射液组AQP3的表达水平低于复方丹参注射液组(P<0.05),但高于U0126组(P<0.05)。结论羊水过少时,复方丹参注射液通过激活MAPK-ERK1/2信号转导通路来调节人羊膜上皮细胞中AQP3的表达水平。

ABSTRACT Objective To investigate the role of m itogen-activated protein kinases （MAPKs）-ex- tracellular signal regulated kinasel/2 （ERKI/2） signal pathway in the regulation of Compound Danshen Injection （CDI） induced AQP3 expression in the human amniotic epithelial cells （hAECs）. Methods hAECs of term pregnancy with normal amniotic fluid volume （AFV） or isolated oligohydramnios were pri- marily cultured. And the cells were equally divided into four groups, i.e., the vehicle control group, the U0126 group, the CDI group, the CDI ＋ U0126 group. The expressions of phosphorylated ERKI/2 （p- ERKl/2）and AQP3 in hAECs were detected using Western blot analysis. Results （1） When compared with the control group, the expression level of p-ERKI/2 in hAECs in those with normal AFV and oligo- hydramnios obviously decreased in the U0126 group （P 〈0.05）. The expression level of p-ERKI/2 could be elevated in the CDI group （P 〈0.05）. The expression level of p-ERKI/2 in hAECs was higher in the CDI ＋U0126 group than in the U0126 group, but lower in the CDI ＋ U0126 group than in the CDI group （P 〈0.05）. （2） There was no obvious change in AQP3 expression in hAECs with normal AFV between the U0126 group and the vehicle control group （P 〉0.05）. There was no statistical difference in the expres-sion level of AQP3 between the CDI group and the U0126 ＋CDI group （P 〉0.05）, but they were higher than those in the vehicle control group （P 〈0.05）. （3） Compared with the vehicle control group, the ex- pression level of AQP3 in hAECs with oligohydramnios significantly decreased in the U0126 group and in- creased in the CDI group （P〈0.05）. The expression level of AQP3 was lower in the U0126 ＋CDI group than in the CDI group, but higher in the U0126 ＋CDI group than in the U0126 group （P 〈0.05）. Conclusion CDI could regulate AQP3 expression level in hAECs with oligohydramnios via activating the MAPK-ERKI/ 2 signal transduction pathway.