复方丹参注射液对JNK信号通路调节人羊膜上皮细胞AQP3表达的影响

Compound Danshen Injection Regulated the Expression of AQP3 in the Human Amnion Epithelium Cells through JNK Signal Pathway

目的探讨复方丹参注射液调节人羊膜上皮细胞中水通道蛋白3(aquaporin 3,AQP3)的表达与c-Jun氨基末端激酶(c-Jun N-terminal kinase,JNK)信号通路的关系。方法分别取足月妊娠的正常羊水和羊水过少的人羊膜上皮细胞进行原代培养。将培养的正常羊水和羊水过少的人羊膜上皮细胞分别分成4个处理组:空白对照组(A组)、SP600125组(B组)、复方丹参注射液组(C组)和SP600125加复方丹参注射液组(D组)。采用cell counting kit-8(CCK-8)法检测各组细胞的活力,Western blot技术分析人羊膜上皮细胞中总JNK、磷酸化JNK(p-JNK)和AQP3蛋白的表达量。结果(1)在正常羊水及羊水过少人羊膜上皮细胞中:4组细胞活力和总JNK蛋白差异均无统计学意义(P>0.05)。p-JNK差异有统计学意义(P<0.05)。与A组比较,p-JNK在B组表达明显下降,而在C组中其表达明显上调(P<0.05)。且p-JNK在D组中的表达较C组明显下降,但高于A组和B组(P<0.05)。与A组比较,在正常羊水人羊膜上皮细胞中,C、D组中AQP3表达明显上调(P<0.05)。但与C组比较,D组AQP3表达差异无统计学意义(P>0.05)。而在羊水过少人羊膜上皮细胞中,B组中AQP3的表达明显下降(P<0.05),而C组中AQP3表达明显上调(P<0.05);D组中AQP3表达低于C组(P<0.05),但高于B组(P<0.05)。结论羊水过少时,复方丹参注射液可通过激活JNK信号通路来调节人羊膜上皮细胞中AQP3表达的水平。

Objective To explore the role of Compound Danshen Injection （CDI） in regulating the expression of aquaporin 3 （AQP3） in human amnion epithelium cells （hAECs）, and to study the relation between c-Jun N-terminal kinase （JNK） signal pathway and AQP3. Methods hAECs were isolated and primarily cultured from term pregnancy with normal amniotic fluid volume and from term pregnancy with oligohydramnios, and then hAECs were further divided into four groups, i.e., the blank control group （A）, the SP600125 group （B）, the CDI group （C）, and the SP600125 ＋CDI group （D）. The cell viability was measured by cell counting kit-8 assay （CCK-8）. The expression of total JNK, phosphorylated JNK, and AQP3 were determined by Western blot. Results （1） In hAECs with normal AFV or with oligohydr- amnios ： There was no statistical difference in the cell viability or the expression of total JNK among the 4 groups （P 〉0.05）. But there was statistical difference in the expression of p-JNK （P 〈0.05）. Compared with A group, the expression of p-JNK was obviously down-regulated in B group, but obviously up-regula- ted in C group （P 〈0.05）. The expression of p-JNK was significantly lower in D group than in C group, but higher than that in A group or B group （P 〈0.05）. The AQP3 expression in the hAECs with normal amniotic fluid volume of C group and D group were higher than that in the A group （P 〈0.05）. However, there was no statistical difference in the AQP3 expression between C group and D group （P 〉0.05）. In hAECs with oligohydramnios, the expression of AQP3 obviously decreased in B group, but up-regulated in C group （both P 〈0.05）. The expression of AQP3 was lower in D group than in C group, but higher than in B group （P 〈0.05）. Conclusion CDI could regulate the AQP3 expression in hAECs with oligohydramnios via activating the JNK signal pathway.