摘要
目的:探讨活血化湿汤靶向TP53对M2巨噬细胞极化与非小细胞肺癌(NSCLC)细胞侵袭的作用。方法:网络药理学分析筛选出活血化湿汤治疗NSCLC的靶点TP53。qRT-PCR和Western blot检测TP53在NSCLC细胞中的表达水平。诱导THP-1细胞为M0巨噬细胞,使用活血化湿汤及TP53处理巨噬细胞,流式细胞术分析CD86、CD206表达,ELISA检测IL-1β、CCL18浓度。使用活血化湿汤、TP53过表达载体、巨噬细胞培养液处理NSCLC细胞,Transwell检测NSCLC细胞侵袭能力。结果:网络药理学分析发现活血化湿汤的靶点和NSCLC的发病风险基因靶点共交集17个基因,STRING分析显示TP53与其他基因的交互作用最强。与BEAS-2B细胞比较,NSCLC细胞中TP53表达增强,而活血化湿汤能抑制TP53表达。活血化湿汤可通过抑制TP53上调巨噬细胞中CD86阳性细胞百分比,下调CD206阳性细胞百分比,提高IL-1β浓度,降低CCL18浓度(均P<0.05)。此外,活血化湿汤能通过抑制TP53表达进而抑制NSCLC细胞的侵袭。结论:活血化湿汤通过抑制TP53来抑制M2巨噬细胞极化,从而抑制NSCLC细胞侵袭。
Objective:To investigate the effects of Huoxue Huashi Decoction targeting TP53 on M2 macrophage polarization and non-small cell lung cancer(NSCLC)cell invasion.Methods:Network pharmacology analysis was used to screen out the target TP53 of Huoxue Huashi Decoction in treatment of NSCLC.Expression level of TP53 in NSCLC cells was detected by qRT-PCR and Western blot.THP-1 cells were induced to M0 macrophages,the macrophages were treated with Huoxue Huashi Decoction and TP53.Expressions of CD86 and CD206 were analyzed by flow cytometry,and concentrations of IL-1βand CCL18 were detected by ELISA.NSCLC cells were treated with Huoxue Huashi Decoction,TP53 overexpression vector and macrophage culture medium,and the invasive ability of NSCLC cells were detected by Transwell.Results:Network pharmacology analysis found that there were 17 genes in intersections between the targets of Huoxue Huashi Decoction and NSCLC risk genes.STRING analysis showed that TP53 had the strongest interaction with other genes.Compared with BEAS-2B cells,expression of TP53 was enhanced in NSCLC cells,while Huoxue Huashi Decoction could inhibit the expression of TP53.Huoxue Huashi Decoction could increase the percentage of CD86-positive cells in macrophages by inhibiting TP53,down-regulate percentage of CD206-positive cells,increase the concentration of IL-1βand decrease the concentration of CCL18(all P<0.05).In addition,Huoxue Huashi Decoction could inhibit the invasion of NSCLC cells by inhibiting expression of TP53.Conclusion:Huoxue Huashi Decoction inhibits the polarization of M2 macrophages by inhibiting TP53 and thus inhibits invasion of NSCLC cells.
作者
高莉萍
赵兰婷
冯倩
石军年
GAO Liping;ZHAO Lanting;FENG Qian;SHI Junnian(Respiratory Medicine,the Second Hospital of Lanzhou University,Lanzhou 730030,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2023年第10期2049-2054,I0002-I0003,共8页
Chinese Journal of Immunology
基金
甘肃省高等学校创新基金项目(2021B-035)。
