温下方调控肿瘤相关巨噬细胞M2型极化抑制肺癌侵袭转移的机制  

Mechanism of Wenxia Formula regulating M2 polarization of TAMs and inhibiting the invasion and metastasis of lung cancer

作　　者：王梦然 毕倩宇 章蓓滢 黄盛琦 季旭明 WANG Meng-ran;BI Qian-yu;ZHANG Bei-ying;HUANG Sheng-qi;JI Xu-ming(School of Basic Medical Sciences,Zhejiang Chinese Medical University,Hangzhou 310053,China;Laboratory of Neuropharmacology and Translational Medicine of Zhejiang Province,Hangzhou 310053,China;Academy of Chinese Medical Sciences,Zhejiang Chinese Medical University,Hangzhou 310053,China)

机构地区：[1]浙江中医药大学基础医学院,杭州310053 [2]浙江省神经药理学与转化研究重点实验室,杭州310053 [3]浙江中医药大学中医药科学院,杭州310053

出　　处：《中华中医药杂志》2022年第5期2799-2804,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.81774198)。

摘　　要：目的:探讨温下方对巨噬细胞极化调控机制及M2型巨噬细胞(M2-TAMs)对A549细胞侵袭转移的影响。方法:动物实验:BALB/c裸鼠随机分为模型组和温下方组,灌胃干预。活体成像分析光子强度;观察肺部病理变化;识别CD68/CD163双标M2-TAMs。细胞实验:佛波酯(PMA)及白介素-4(IL-4)诱导M2-TAMs;流式细胞术检测M2-TAMs标志蛋白CD163表达;PCR检测温下方对M2-TAMs相关基因mRNA表达的影响;划痕和Transwell检测温下方对M2-TAMs促A549细胞侵袭转移的影响;Western blot和PCR检测非受体型酪氨酸蛋白激酶3(J AK3)、信号转导及转录激活因子6(ST AT6)和过氧化物酶增殖物激活受体γ(PP A Rγ)蛋白及mRNA表达。结果:动物实验显示,与模型组比较,温下方在第28天~35天有效降低转移灶光子强度(P<0.05,P<0.01);温下方有效减轻肺部病理损伤;温下方降低转移瘤中M2-TAMs特异性表型CD68和CD163双阳表达。细胞实验显示,成功诱导的M2-TAMs高表达CD163;与M2组比较,温下方显著抑制M2-TAMs标志物Arg-1、Fizz-1、C D206 m R NA表达(P<0.01);与M2组比较,M2+WC F组显著降低M2-T A M s促A549细胞的迁移和侵袭能力(P<0.01);与M2组相比较,温下方有效降低p-JAK3、p-STAT6和PPARγ蛋白及JAK3、STAT6、PPARγmRNA表达(P<0.05,P<0.01)。结论:温下方可能通过调控JAK3/STAT6/PPARγ信号通路,抑制巨噬细胞M2极化,降低A549细胞侵袭转移。Objective:To investigate the regulation mechanism of Wenxia Formula on macrophage polarization and the effect of M2 macrophages(M2-TAMs)on the invasion and metastasis of A549 cells.Methods:Animal experiments:BALB/c nude mice were randomly divided into the model group and the Wenxia Formula group with intragastric administration.Living image was used to analyse photon intensity;The pathological changes of lung were observed;CD68/CD163 double labeled M2-TAMs was identified.Cell experiments:PMA and IL-4 were used to induce M2-TAMs;The expression of M2-TAMs marker protein CD163 was detected by flow cytometry;The effect of Wenxia Formula on the mRNA expression of M2-TAMs related genes was detected by PCR;The effects of Wenxia Formula on the invasion and metastasis of A549 cells promoted by M2-TAMs were detected by scratch and Transwell;JAK3/STAT6/PPARγprotein and mRNA expression were detected by Western blot and PCR.Results:Animal experiments showed that compared with the model group,the photon intensity of metastasis in Wenxia Formula group was effectively reduced from day 28 to 35(P<0.05,P<0.01);Wenxia Formula effectively reduced the pathological injury of the lung;Wenxia Formula decreased the double positive expression of M2-TAMs specific phenotypes CD68 and CD163 in metastatic tumors.Cell experiment showed that M2-TAMs successfully induced high expression of CD163;Compared with M2 group,Wenxia Formula significantly inhibited the Arg-1,Fizz-1,CD206 mRNA expression of M2-TAMs markers(P<0.01);Compared with M2 group,M2+WCF group significantly decreased the migration and invasion ability of M2-TAMs in A549 cells(P<0.01);Compared with M2 group,Wenxia Formula effectively reduced p-JAK3,p-STAT6 and PPARγprotein and JAK3,STAT6,PPARγmRNA expression(P<0.05,P<0.01).Conclusion:Wenxia Formula could regulate JAK3/STAT6/PPARγsignal pathway,inhibit M2 polarization of macrophages and reduce the invasion and metastasis of A549 cells.

关 键 词：温下方 肿瘤相关巨噬细胞 M2型极化 肿瘤微环境 肺癌 侵袭 转移 

分 类 号：R285.5[医药卫生—中药学]