ABSTRACT
Objective: To develop an UPLC-Q-TOF-MS method for the determination of seven components (naringin, hesperidin, strychnine, brucine, agrimol B, wogonin, and protocatechuic acid) in Pingxiao Tablets. Methods: The analysis was performed on a Waters Acquity BEH C18 column (100 mm×2.1 mm, 1.7 μm) with the mixture of acetonitrile-water-formic acid as mobile phase, and the flow rate was 0.45 mL/min; MS scanning mode: negativion; Sample volume: 2.0 μL. Results: The linear relationship between the concentration and peak areas of the seven compounds were all linear (r>0.9996). The average recoveries (n=6) were between 99%-101%. Conclusion: This method is simple, accurate, and reliable to determine the seven contents in Pingxiao Tablets for the quality control.
ABSTRACT
OBJECTIVE: To investigate the effect and mechanism of allomatrine in proliferation and invasion in vitro inhibition of human lung cancer A549 cell line. METHODS: After treatment with allomatrine, MTS assay was employed to determine the proliferation of cancer cells, flow cytometry to determine the apoptosis rate, cell cycle distribution and intracellular ROS production, transwell assay to determine the cell invasion potential in vitro, adhesion assay to determine the cell adhesion potential in vitro, realtime PCR and Western blotting assay to determine the expression of apoptosis related gene Survivin, Bcl-2 and caspase-3/8, the cell cycle related protein CDK-2, adhesion molecule CD44, matrix metalloproteinases MMP-2/9 and the phosphorylation of AKT, report gene assay to determine the transcription activity of NF-KB and the activity of ubiquitin proteasome. RESULTS: Allomatrine significantly inhibited the proliferation and invasion in vitro of A549 cells, flow cytometry showed that apoptosis rate and ROS production was increased and the cell cycle was halted by G2/M phase, the expression of pro-apoptosis gene caspase-3/8 was upregulated while the anti-apoptosis proteins Survivin and Bcl-2 was downregulated, the expression of CDK-2, CD44 and MMP-2/9 was downregulated, and the phosphorylation of AKT was downregulated, the transcription activity of NF-KB and the activity of ubiquitin proteasome were inhibited after Allomatrine treatment. CONCLUSIONS: Allomatrine was able to inhibit proliferation and invasion in vitro of human lung cancer A549 cell line by inducing ROS production, promoting apoptosis, arresting cell cycle, inhibiting ubiquitin proteasome and regulating tumor related gene expression.