摘要
目的探究芫花醇提物对骨髓增生异常综合征细胞株的药效学作用。方法使用不同浓度芫花醇提物处理SKM-1和MUTZ-1细胞株72 h,CCK8检测细胞在0,24,48和72 h时间点增殖情况。在后续的实验中,使用浓度为2,5和10 mg/mL的芫花醇提物处理SKM-1和MUTZ-1细胞24 h。流式细胞术检测细胞凋亡;Elisa检测细胞IL-6及TNF-α含量变化;q-RT PCR检测NF-κB表达变化;Western blot检测细胞核和浆中NF-κB蛋白表达,检测Caspase3和Survivin蛋白表达。结果芫花醇提物可以剂量依赖性地抑制SKM-1和MUTZ-1细胞增殖,并促进SKM-1和MUTZ-1细胞凋亡。芫花醇提物可以抑制MUTZ-1细胞IL-6及TNF-α炎症因子的表达含量,抑制NF-κB mRNA表达以及细胞核中NF-κB蛋白含量,同时抑制survivin蛋白的表达并促进Caspase3的表达。另外,MUTZ-1细胞中NF-κB mRNA及核中NF-κB蛋白受外源TNF-α的诱导,但这种现象受到芫花醇提物处理的抑制。结论芫花醇提物可以抑制SKM-1和MUTZ-1细胞的增殖并促进SKM-1和MUTZ-1细胞的凋亡,其作用机制与减少炎症因子及核NF-κB含量蛋白有关。
Objective To discover the pharmacodynamic effects of ethanol extracts from daphne genkwa against myelodysplastic syndrome in human cells.Methods The cell line SKM-1 and MUTZ-1 were treated by different concentration of ethanol extracts from daphne genkwa for 72 h,the proliferation of cells was tested using CCK8 assay kits at 0,24,48 and 72 h after treatment.In the follow-up experiments,the SKM-1 and MUTZ-1 cells were treatment with the ethanol extracts from daphne genkwa at the concentration of 2,5 and 10 mg/mL for 24 h.The apoptosis rate of cells was tested using flow cytometry.The content of inflammatory factors IL-6 and TNF-αwas tested using ELISA kits.The mRNA expression of NF-κB was determined by q-RT PCR.The protein levels of NF-κB(in cytoplasm and nucleus),Caspase3 and Survivin were detected by western blot.Results The proliferation of SKM-1 and MUTZ-1 cells was inhibited by ethanol extracts from daphne genkwa in a dose-dependent manner,and the apoptosis rate of cells was increased by treatment with ethanol extracts from daphne genkwa.Further,the inflammatory factors IL-6 and TNF-αcontents in MUTZ-1 were decreased by treatment with ethanol extracts from daphne genkwa.Moreover,the mRNA expression of NF-κB as well as protein levels of NF-κB(in nucleus),Caspase3 and Survivin were decreased by treatment with ethanol extracts from daphne genkwa.In addition,the protein level of NF-κB(in nucleus)was induced by exogenous TNF-α,and this phenomenon was ameliorated by ethanol extracts from daphne genkwa.Conclusion Ethanol extracts from daphne genkwa could inhibit the proliferation and promote the apoptosis of SKM-1 and MUTZ-1 cells,the specific mechanisms were related to the decrease of inflammatory factors and protein level of NF-κB in nucleus.
作者
陈蔚
姚永华
胡慧菁
CHEN Wei;YAO Yong-hua;HU Hui-jing(Department of Pharmacy,Shidong Hospital of Yangpu District,Shanghai 200438,China;不详)
出处
《医药论坛杂志》
2020年第7期79-84,共6页
Journal of Medical Forum
基金
上海市杨浦区卫计委中医药专项课题(YP17ZM03)
上海市综合医院中西医结合专项(ZHYY-ZXYJHZX-2-19)。
关键词
芫花素醇提物
MDS
炎症因子
细胞凋亡
Ethanol extracts from daphne genkwa
MDS
Inflammatory factors
Cell apoptosis
