摘要
目的研究胃复春胶囊对人胃癌细胞株MKN-49P的作用,为胃复春胶囊在抗消化道肿瘤方面的应用提供新的依据。方法将SD大鼠随机分为2组,即对照组、胃复春胶囊组。胃复春胶囊组取胶囊内容物,以生理盐水混悬,按300g/(kg·d)剂量ig给药,对照组大鼠ig 2 mL生理盐水,共5 d。末次给药后2 h,在无菌条件下,ip 10%水合氯醛(350 mg/kg)麻醉,腹主动脉取血,分离血清,经灭活处理后,置于-20℃冰箱保存备用。用不同质量浓度(0、2.5、5、10、20、30、50、80、100μg/mL)的胃复春胶囊含药血清处理MKN-49P细胞,采用MTT法检测细胞活力。加入各质量浓度(25、50、100μg/m L)胃复春胶囊含药血清,流式细胞术Annexin V/PI双染色检测细胞凋亡情况,Hoechst染色法检测细胞形态,细胞划痕实验测定细胞的迁移能力,Western blotting法检测信号传导与转录激活因子(STAT)蛋白、核转录因子-κB(NF-κB)信号通路及与细胞凋亡相关的蛋白表达情况。结果胃复春胶囊含药血清能够剂量和时间相关地降低MKN-49P细胞的活力,并可诱导MKN-49P细胞凋亡。Hoechst染色显示细胞膜通透性增强;细胞划痕实验结果显示,与对照组(0μg/mL)相比,不同浓度胃复春胶囊含药血清(25、50、100μg/mL)可浓度相关性地抑制MKN-49P细胞的迁移。MKN-49P细胞经过25、50、100μg/mL胃复春胶囊含药血清分别处理24 h后,MKN-49P细胞中STAT3的磷酸化减弱,NF-κB信号通路相关的蛋白表达量下调,抗凋亡蛋白表达量下降而促凋亡蛋白表达量增加。结论胃复春胶囊含药血清能抑制胃癌细胞的增殖、诱导细胞凋亡,并显著降低细胞迁移能力,其作用机制推测与NF-κB信号通路有关。
Objective To study the effect of Weifuchun Capsules on human gastric cancer cell line MKN-49P,and to provide a new basis for the application of Weifuchun Capsules in anti-digestive tract tumor.Methods SD rats were randomly divided into two groups:control group and Weifuchun Capsules group.Weifuchun Capsules group was given the Weifuchun Capsules contents,suspended with normal saline,ig at 300 g/(kg·d)dose,and control group was ig 2 mL normal saline,for a total of 5 d.2 h after the last administration,under aseptic conditions,10%chloral hydrate(350 mg/kg)was injected intraperitoneally for anesthesia.Blood was taken from abdominal aorta and the serum was separated.After inactivation treatment,the serum was stored in a refrigerator at-20℃for future use.MKN-49P cells were treated with Weifuchun Capsules containing serum at different concentrations(0,2.5,5,10,20,30,50,80,and 100μg/mL).Cell viability was detected by MTT assay.Add the drug containing serum of Weifuchun Capsules at different concentrations(25,50,100μg/mL),apoptosis was detected by Annexin V/PI double staining by flow cytometry.Cell morphology was detected by Hoechst staining.The cell migration ability was measured by cell scratch assay.The expression of signal transduction and transcription activator(STAT)protein,nuclear transcription factor-κB(NF-κB)signaling pathway and apoptosisrelated proteins were detected by Western blotting.Results The serum containing Weifuchun Capsules could decrease the viability of MKN-49P cells,and induce apoptosis of MKN-49P cells in a dose and time dependent manner.Hoechst staining showed enhanced cell membrane permeability.Compared with control group(0μg/mL),different concentrations of Weifuchun Capsules containing serum(25,50,and 100μg/mL)could inhibit the migration of MKN-49P cells in concentration-dependent manner.After MKN-49P cells were treated with 25,50,and 100μg/mL serum containing Weifuchun Capsules for 24 h,the phosphorylation of STAT3were decreased.The protein expression of NF-κB signaling pathway was down-regulated.The expression of anti-apoptotic protein decreased while that of pro-apoptotic protein increased.Conclusion The drug-containing serum of Weifuchun Capsules can inhibit the proliferation and induce apoptosis of gastric cancer cells,and significantly reduce the cell migration ability.The mechanism of action is speculated to be related to NF-κB signaling pathway.
作者
祁大庆
陈琳慧
潘海春
万林春
QI Da-qing;CHEN Lin-hui;PAN Hai-chun;WAN Lin-chun(Hangzhou Huqing Yutang Pharmaceutical Co.,Ltd.,Hangzhou 310000,China;Jiangxi Institute For Drug Control,Nanchang 330029,China)
出处
《现代药物与临床》
CAS
2022年第6期1175-1181,共7页
Drugs & Clinic