爱康方含药血清对人肺腺癌A549细胞PI3K、Akt及p-Akt表达的影响

Effects of serum containing Ai Kang recipe on the expression of PI3K,Akt and p-Akt in human lung adenocarcinoma A549 cells

目的初步观察爱康方20%浓度最佳含药血清对A549细胞PI3K、Akt及p-Akt表达的影响,探讨其细胞的凋亡作用是否与磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/Akt)通路有关。方法参照课题组前期制备含药血清方法制备爱康方含药血清,将SD大鼠40只分为4组:生理盐水对照组(NS组),爱康方低、中、高剂量组,每组10只,制备含药血清。将SD大鼠分组对应所制备的含药血清,调配成20%最佳含药血清一一对应干预A549细胞,再补充只加DMEM培养基的细胞作为空白对照组。采用Western blot检测各组细胞PI3K、Akt、p-Akt表达。结果对于PI3K,培养24 h后,与NS组和空白对照组比较,除低剂量组,中、高剂量PI3K蛋白表达水平下调(P<0.05);培养48 h后,与空白对照组和NS组比较,低、中、高剂量组PI3K蛋白表达水平均下调(P<0.05)。对于Akt,培养24 h后,与空白对照组和NS组比较,中、高剂量组Akt蛋白表达水平下调(P<0.05);培养48 h后,与空白对照组和NS组比较,中、高剂量组Akt蛋白表达水平下调(P<0.05)。对于p-Akt,培养24 h后,与空白对照组和NS组比较,高剂量组p-Akt蛋白表达水平降低(P<0.05)。结论爱康方含药血清能下调A549细胞PI3K、Akt、p-Akt蛋白表达,推测其发挥的凋亡作用可能与PI3K/Akt信号通路有关。

Objective To preliminarily observe the effect of the 20%Ai Kang recipe of the best drug-containing serum on the expression of PI3K,Akt and p-Akt in A549 cells,and to explore the relationship between cell apoptosis and PI3K/Akt pathway.Methods Forty SD rats were divided into the control group(NS group),the low,medium and high dose of Ai Kang recipe groups(the low,medium and high group,10 in each group).The corresponding drug-containing serum of each group of rats was adjusted to 20%concentration of the best drug-containing serum,then the best drug-containing serum was used to intervened the A549 cells.The A549 cells in the NS group were intervened with physiological saline,and then supplemented with the blank control group which interval cells with only DMEM medium.The expression of PI3K and Akt of each groupwas detected by Western blot.Results For PI3K,except for the low-dose group,the middle and high dose groups were lower than the NS group and the blank control group after 24 hours of culture(P<0.05);The low,medium and high dose groups were lower than the NS group and the blank control group after 48 hours of culture(P<0.05).For Akt,the high-dose group was significantly reduced when compared with the blank control group and the NS group after 24 hours of culture(P<0.05),compared with the blank control group and NS group,the high and medium dose groups were significantly down-regulated(P<0.05).For p-Akt,the high-dose group was significantly reduced when compared with the blank control group and the NS group after 24 hours of culture(P<0.05).Conclusion Ai Kang recipe can down-regulate the expression of PI3K,Akt and p-Akt in A549 cells,which suggests that the apoptotic effect may be related to the PI3K/Akt signaling pathway.