摘要
目的:探究南方红豆杉水提物含药血清联合吉非替尼含药血清对A549细胞株增殖的影响。方法:新西兰纯家兔26只,雌雄各半,随机分成6组,即对照组(0.9%氯化钠溶液),南方红豆杉水提物低剂量组(560mg·kg^-1·d^-1),南方红豆杉水提物中剂量组(1120m g·k g^-1·d^-1),南方红豆杉水提物高剂量组(2240m g·k g^-1·d^-1),吉非替尼标准剂量组(16.35m g·k g^-1·d^-1),南方红豆杉水提物高剂量+吉非替尼组(2240m g·k g^-1·d^-1+16.35mg·kg^-1·d^-1),连续灌胃7d后,末次灌胃1h后,无菌心脏采血,制备血清。取10%各组家兔血清作用于A549细胞株24、48、72h后,运用CCK-8法检测各组细胞增殖情况;Western Blot检测ERK和JAK/STAT3通路相关蛋白表达;RT-PCR检测ERK和JAK/STAT3 mRNA表达情况。结果:CCK-8检测:在作用时间相同时,与南方红豆杉水提物低剂量组比较,中、高剂量组对A549细胞生长抑制率升高(P<0.05);与吉非替尼组比较,南方红豆杉水提物+吉非替尼组对A549细胞生长抑制率显著升高(P<0.05);Western Blot检测显示与对照组和吉非替尼组比较,联合组能显著下调p-EGFR、p-ERK1/2、p-JAK2、p-STAT3等蛋白表达(P<0.01);在RT-PCR检测中,与对照组和吉非替尼组比较,南方红豆杉组和联合组中ERK2、STAT3 mRNA表达显著下调(P<0.01)。结论:南方红豆杉水提物含药血清能增强吉非替尼对A549细胞增殖的抑制作用,其机制与抑制EGFR下游ERK及PI3K/AKT信号通路有关。
Objective:To explore the effect of drug-containing serum of Taxus chinensis var.mairei combined with gefitinib-containing serum on proliferation of A549 cell line.Methods:Twenty-six New Zealand pure rabbits,male and female,were randomly divided into 6 groups,namely the control group(normal saline),the low dose group of Taxus chinensis(560mg·kg^-1·d^-1),and the middle dose group of Taxus chinensis(1120mg·kg^-1·d^-1),high dose group of Taxus chinensis(2240mg·kg^-1·d^-1),standard dose group of gefitinib(16.35mg·kg^-1·d^-1),group of Taxus chinensis+gefitinib(2240mg·kg^-1·d^-1+16.35mg·kg^-1·d^-1),7 days after continuous gavage,1h after the last gavage,blood was collected from the sterile heart to prepare serum.The serum of 10%rabbits was treated with A549 cells for 24h,48h and 72h.The proliferation of each group was detected by CCK-8 method.The expression of ERK and JAK/STAT3 pathway-related proteins was detected by Western Blot.The expression of ERK and JAK/STAT3 mRNA was detected by RT-PCR.Results:CCK-8 detection:When the time of action was the same,the growth inhibition rate of A549 cells was higher in the middle and high dose groups than in the low dose group of Taxus chinensis(P<0.05).Compared with the gefitinib group,the growth inhibition rate of A549 cells was significantly increased in the Taxus chinensis+Gefitinib group(P<0.05).Western Blot showed that compared with the control group and gefitinib group,the combination group could significantly down-regulate the expression of p-EGFR,p-ERK1/2,p-JAK2,p-STAT3 and other proteins(P<0.01).In the RTPCR assay,the expression of ERK2 and STAT3 mRNA was significantly down-regulated in the Taxus chinensis group and the combination group compared with the control group and the gefitinib group(P<0.01).Conclusion:The drug-containing serum of Taxus chinensis var.mairei can enhance the inhibitory effect of gefitinib on the proliferation of A549 cells,which may be through inhibiting ERK and PI3K/AKT signaling pathways downstream of EGFR.
作者
陈婷
张郜晨茜
戴淑颖
舒琦瑾
CHEN Ting;ZHANG Gao-chen-xi;DAI Shu-ying;SHU Qi-jin(First College of Clinical Medicine,Zhejiang Chinese Medical University,Hangzhou 310053,China;Department of Oncology,The First Affiliated Hospital of Zhejiang Chinese Medical University,Hangzhou 310006,China)
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2020年第2期655-659,共5页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
国家自然科学基金项目(No.81173247)。
