(+)-corynoline Regulates the Proliferation,Stemness and Apoptosis of Triple Negative Breast Cancer Cells / 中国医学科学院学报

Objective To explore the performance and mechanism of(+)-corynoline in treating triple negative breast cancer MDA-MB-436 cells and thus provide an option for the development of drugs against this cancer. Methods The viability,proliferation,apoptosis and migration/invasion of MDA-MB-436 cells treated with(+)-corynoline were detected by CCK-8 assay,colony formation assay,flow cytometry and Transwell assay,respectively.Furthermore,Western blotting was employed to determine the expression of related proteins,and RNA-Seq was performed for the MDA-MB-436 cells treated with(+)-corynoline. Results (+)-corynoline inhibited the proliferation and stemness and promoted the apoptosis of MDA-MB-436 cells.Further,(+)-corynoline may activate the oxidative phosphorylation pathway to play a role in inhibiting triple negative breast cancer. Conclusion (+)-corynoline can inhibit triple negative breast cancer cells,which helps to address the poor efficacy of existing chemotherapeutics and facilitate the development of drugs against this cancer.

Pharmacokinetics of active ingredients of Pudilan Xiaoyan Oral Liquid in rats and effect on liver drug-metabolizing enzymes / 中国中药杂志

This study is aimed to establish a method for the determination of baicalin,baicalin and purpurin in the plasma of rats after oral administration of Pudilan Xiaoyan Oral Liquid( PDL) by using liquid chromatography-mass spectrometry( LC-MS),analyze the pharmacokinetics of three components in rats,and investigate the effects of PDL on drug-metabolizing enzymes in rat liver. C18 column was used for liquid chromatography separation,with acetonitrile-water( containing 0. 2% formic acid) as the mobile phase for gradient elution. The mass spectrometry was detected by electrospray ion source( ESI) under multi-reaction monitoring mode( MRM),as well as positive and negative ion alternating mode. Plasma sample collection was performed by using an automatic blood collection meter for small animals. The pharmacokinetic parameters were calculated by Win Nonlin software. The total protein concentration of rat liver microsomes and the total enzyme content of CYP450 were determined by BCA method and spectrophotometry respectively. The methodological study in terms of linear range,recovery rate,precision and sample stability,was used to confirm that the LC-MS analysis method established in this experiment was simple,exclusive,accurate and reliable,and can meet the requirement of determining the content of baicalin,oroxindin and corynoline in plasma after PDL administration in rats. The drug-time curve showed that baicalin and oroxindin had a bimodal phenomenon,and the pharmacokinetic parameters indicated that baicalin,oroxindin and corynoline in PDL had certain drug-like properties. After 7 consecutive days of PDL administration,the rat liver coefficient,total liver microparticle protein and CYP450 enzyme content were increased,but there was no significant difference,indicating that PDL was less likely to develop drug-drug interaction based on CYP enzyme. The results of this experiment can provide reference for the research on in vivo efficacy and drug interaction of PDL as well as on its clinical application.

Anti-inflammatory effect of corynoline isolated from Corydalis bungeana Turcz / 中国药科大学学报

To isolate corynoline from Corydalis bungeana Turcz.and study its anti-inflammatory mechanism via TLRs/NF-κB signal pathway.Corynoline was extracted by 80％ ethanol and purified by silicagel column chromatography.The structure and purity of corynoline was determined by UPLC,MS,1H NMR and 13C NMR.In the course of experiment,the cytotoxicity of corynoline was evaluated by MTT assay.And the inflammation model was established by RAW264.7 macrophages induced by lipopolysaccharide(LPS),which was intervened by coryno line.The expression levels of TLR4,TLR2 and nuclear transcription factor-κB(NF-κB) signaling pathways related proteinsin RAW264.7 macrophages were detected by Western blot.Furthermore,the expressionof NF-κB p65 mRNA and nuclear p65 were determined by the real-time fluorescence quantitative PCR(RT-qPCR) and Western blot.Results showed that 5-40 μmol/L corynoline reduced the expression level of TLR4 and TLR2,and inhibited the phosphorylation level of IκBα and the phosphorylation and nuclear translocation of p65 at gene and protein levelin a dose-dependent manner in LPS-induced RAW264.7 cells.This study indicated the protective effect of corynoline on LPS-induced RAW264.7 macrophages may be related with the inhibition of TLRs/NF-κB inflammatory signaling pathway.