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目的:观察加味人参乌梅汤对Caco-2脱水模型AQP4基因表达的影响,初步探讨其生津止泻的作用机制。方法:常规方法建立Caco-2细胞高渗性脱水细胞模型,以不同浓度的加味人参乌梅汤处理,光镜观察细胞形态,MTT法检测细胞活力,并采用RT-PCR方法检测各组细胞AQP4基因表达水平。结果:高渗培养基处理Caco-2细胞后,细胞呈现脱水现象,高渗培养液培养3h、6h、12h、24h后脱水模型组的A值均明显低于对照组,AQP4基因表达较正常对照组亦显著降低(P<0.05);用含有不同浓度加味人参乌梅汤的培养液处理细胞,细胞脱水程度明显减缓,且表现出一定的量效关系,各时间点A值均较模型组显著增高,AQP4基因表达较模型组显著增高(P<0.05或0.01)。结论:加味人参乌梅汤能显著对抗Caco-2细胞的高渗性脱水损伤,增强AQP4基因表达可能是加味人参乌梅汤益气生津止泻效应的作用靶点之一。
OBJECTIVE: To observe the effect of Jiawei Renshen Wumei Decoction on the expression of AQP4 gene in Caco-2 dehydration model, and to explore the mechanism of Shengjin Zhixie. METHODS: The hyperosmotic dehydration cell model of Caco-2 cells was established by routine methods. The cells were treated with different concentrations of flavored ginseng and Wumei Decoction. The cell morphology was observed by light microscope. The viability of cells was detected by MTT assay. The AQP4 gene was detected by RT-PCR. The expression level. RESULTS: After Caco-2 cells were treated with hypertonic medium, the cells showed dehydration. After 3h, 6h, 12h and 24h of hyperosmotic medium culture, the A values of the dehydration model group were significantly lower than those of the control group. The expression of AQP4 gene was higher than that of normal controls. The group also significantly reduced (P<0.05); treatment of cells with different concentrations of flavored ginseng Wumei Decoction treated cells significantly slowed the dehydration of the cells, and showed a certain dose-effect relationship, and the A value at each time point was significantly higher than the model group. The expression of AQP4 gene was significantly higher than that of the model group (P<0.05 or 0.01). Conclusion: Modified ginseng Wumei Decoction can significantly inhibit hypertonicity of Caco-2 cells. Enhancing the expression of AQP4 gene may be one of the targets for improving the diarrhea effect of ginseng and Wumei Decoction.