臭牡丹苯乙醇苷类成分的鉴定及体外抗人肺腺癌A549细胞的机制研究

Identification of phenylethanoid glycosides from Clerodendrum bungei and its mechanism of anti-human lung adenocarcinoma A549 cell in vitro

臭牡丹提取物具有良好的抗肿瘤的作用,为了鉴定臭牡丹提取物的成分及探究其对A549细胞体外抑制的影响,采用超高效液相色谱-质谱联用技术、CCK-8、细胞划痕法、Transwell、Western blot、免疫荧光及流式细胞术等方法进行检测。鉴定了臭牡丹提取物中34个苯乙醇苷类化合物,其中类叶升麻苷含量为32.95%,异类叶升麻苷5.49%。体外实验表明,臭牡丹提取物能抑制A549细胞的增殖、迁移和侵袭,且IC 50为0.125 mg/mL,还能下调A549细胞中EphA2、p-AKT、mTOR、p-mTOR、p-GSK-3β和β-catenin的表达,上调EphrinA1的表达,减少EphA2在A549细胞膜与膜质上的表达,并将A549细胞周期阻滞于G1/S期。本研究证实臭牡丹地上部分富含苯乙醇苷类成分,尤以类叶升麻苷和异类叶升麻苷为多,且可通过调控EphA2/Akt/mTOR通路达到抗肺癌作用。

Clerodendrum bungei extract has a good anti-tumor effect.In order to identify the ingredients of the C.bungei extract and explore its impact on the A549 in vitro,the ultra-high-efficiency liquid chromatography-mass spectrometry technology,CCK-8,cell scratch,Transwell,Western blot,immunofluorescence and flow cytometry methods were used to detect.The 34 phenylethanoid glycosides compounds of the C.bungei extract were identified,of which the content of acteoside was 32.95%,and isoacteoside was 5.49%.The C.bungei extract can inhibit the proliferation,migration and invasion of A549 cells in vitro,and the IC 50 is 0.125 mg/mL.In addition,the C.bungei extract can reduce the expression of EphA2,P-AKT,mTOR,P-mTOR,P-GSK-3β,andβ-cateninin in A549 cells,increase the expression of EphrinA1,reducing the expression of EphA2 in A549 cell membrane and membrane quality,and blocke the A549 cell cycle to the G1/S stage.This study has confirmed that the C.bungei is rich in phenylethanoid glycosides,especially acteoside and isoacteoside,and can achieve anti-lung cancer effect by regulating the EphA2/AKT/mTOR pathway.