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目的:探讨中药提取成份小檗碱、薯蓣皂苷对滋养细胞内糖代谢相关蛋白分子IRS-1,P1-3K,Glut1,PPARγ表达的调控。方法:人早期妊娠绒毛滋养层细胞体外培养;利用Wortmaninn(WT)进行刺激,诱导细胞糖代谢障碍;采用中药提取成份小檗碱、薯蓣皂苷进行干预,设立曲格列酮(T),二甲双胍(R)为对照药物。利用RT-PCR检测基因表达;Western blot检测蛋白水平的表达;采用激光共聚焦显微技术(LSM)对相关蛋白分子的表达进行定位。结果:①WT诱导前后滋养细胞内葡萄糖代谢发生异常(P<0.05);②WT阻断PI-3K亚基P110的mRNA90.2%的表达;③WT阻断后,IRS-1,PI-3K(P85),PPARγ的mRNA表达下调;④小檗碱可以明显提高IRS-1、P85、PPARγmRNA表达,对IRS-1和PPARγ的调节与曲格列酮相似,药物干预前后分别增加23.94%,113.9%;在P85的调节上明显优于其他组药物,mRNA上调64.41%。蛋白表达与mRNA表达趋势相同,但调控水平不明显。结论:①小檗碱对多种糖代谢相关蛋白因子有调控作用,这可能是改善细胞糖代谢障碍的分子机制;薯蓣皂苷对滋养细胞糖代谢的调节作用较弱;②药物对蛋白表达的调节慢于基因的调节,要提高功能蛋白在细胞糖代谢中的作用是一个相对缓慢的过程。
Objective: To investigate the regulation of berberine and dioscin on the expression of IRS-1, P1-3K, Glut1, and PPARγ in trophoblast cells. METHODS: Human villous trophoblastic cells were cultured in vitro in vitro; Wortmaninn (WT) was used to stimulate the cells to induce glucose metabolism disorder; Traditional Chinese medicine was used to extract berberine and dioscin, and troglitazone (T) and metformin were established. R) is a control drug. RT-PCR was used to detect gene expression. Western blot was used to detect the expression of protein level. Laser confocal microscopy (LSM) was used to locate the expression of related protein molecules. RESULTS: 1The abnormal glucose metabolism in trophoblasts was observed before and after WT induction (P<0.05); 2WT blocked the expression of 91.2% of P110 mRNA in PI-3K subunits; after IWT blocking, IRS-1 and PI-3K (P85) The mRNA expression of PPARγ was down-regulated.4 Berberine could significantly increase the mRNA expression of IRS-1, P85, and PPARγ. The regulation of IRS-1 and PPARγ was similar to that of troglitazone, which increased by 23.94% and 113.9% before and after drug intervention. The regulation of P85 was significantly better than that of other groups of drugs, and the mRNA was upregulated by 64.41%. Protein expression and mRNA expression trends are the same, but the regulation level is not obvious. Conclusion:1 Berberine has a regulation effect on many glycometabolism related protein factors, which may be the molecular mechanism to improve the glycometabolism disorders of cells; diosgenin has a weaker regulation effect on glycogen metabolism of trophoblasts; 2 regulation of protein expression by drugs Slower than the regulation of genes, it is a relatively slow process to increase the role of functional proteins in cellular glucose metabolism.