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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 investigate the relationship between the expression of aquaporin 3 (AQP3) and the c-Jun N-terminal kinase (JNK) signaling pathway in amnion epithelial cells. Methods The normal amniotic fluid and oligohydramnios of human amniotic membrane epithelial cells were used for primary culture. Normal amniotic fluid and amniotic membrane epithelial cells with too few amniotic fluid were divided into four groups: blank control group (A group), SP600125 group (B group), compound Salvia miltiorrhiza injection group (C group) and SP600125 plus compound salvia miltiorrhiza Injection group (group D). Cell viability was measured by cell counting kit-8 (CCK-8). The expression of total JNK, phosphorylated JNK (p-JNK) and AQP3 in human amniotic epithelial cells were analyzed by Western blot. Results (1) In normal amniotic fluid and amniotic fluid oligo human amniotic epithelial cells, there was no significant difference in cell viability and total JNK protein between the 4 groups (P> 0.05). p-JNK difference was statistically significant (P <0.05). Compared with group A, the expression of p-JNK in group B was significantly decreased, while in group C, the expression of p-JNK was significantly increased (P <0.05). The expression of p-JNK in group D was significantly lower than that in group C, but higher than that in group A and group B (P <0.05). Compared with group A, the expression of AQP3 in group C and group D was significantly increased in normal amniotic fluid epithelial cells (P <0.05). However, compared with group C, there was no significant difference in AQP3 expression in group D (P> 0.05). However, the expression of AQP3 in group B was significantly lower than that in group C (P <0.05), while the expression of AQP3 in group C was significantly lower than that in group C (P <0.05) <0.05), but higher than that in group B (P <0.05). Conclusions When oligohydramnios, Fufang Danshen injection can regulate the expression of AQP3 in human amniotic epithelial cells by activating JNK signaling pathway.