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 discuss the regulation of berberine and dioscin extracted from traditional Chinese medicine in the expression of protein moleculer related with glucose metabolism in trophoblast cells ,such as IRS-1,P1-3K,Glutl, PPAR γ. Methods : To culture the chorion trophoblast cell in the early pregnancy, to induce cells to suffer glucose metabolism obstacle with the use of WortmaninnTake advantage of berberine and dioscin extracted from the Chinese traditional medicine with intervention, simultaneously set the troglitazone (T)and dimthyl(R) for the control group. Detect the gene expression with the use of RT- PCT,simutaniously detect the protein expression in molecular level, Examine the expression in the protein level with the technique of Western blot in combination with the technique of LSM for the expression location of protein moleculer related.Results: ①With the induction of WT, the glucose metabolism inside the tropholast cells becomes abnormal, compared with normal (P〈0.05); ②The subunit of PI-3K is P110 induced mRNA expression by WT decreased 90.2%.③With wortmannin blocking IRS-1, phosphatidylinositol 3 - kinase (P85) and PPAR γ mRNA expression reduced.④Berberine can significantly improve the IRS, P85, PPAR γ, mRNA expression. Berberine is similar to troglitazone on the regulation of IRS-1 and PPAR γ,and respectively increased by 23.94% and 113.9% with intervention ; P85 mRNA elevated 64.41%, which is obviously superior to other drugs;Protein and mRNA expression were in the same trend, but the level of regulation is not significant.Conlusion: ①Berberine plays an important role in regulation of various glucose metabolism associated with protein molecule, which may improve glucose metabolism disorder in the intracellular molecular mechanisms; Dioscin is weak in regulation of the glucose metabolism of trophoblast;②The durgs in regulation of the protein expression is slower than that of gene, which suggests that the transition of improving the level of function pr