熊雨 刘洁 邹攀 苏芮 钟文良 江志超 王贤文 何迎春

〔摘要〕 目的 探討益气解毒方中苯丙素类化合物异欧前胡素、阿魏酸、绿原酸对人鼻咽癌CNE2细胞增殖效应的抑制作用及机制研究。方法 使用实时无标记细胞功能分析技术（RTCA）动态监测不同浓度的3种苯丙素类化合物对离体人鼻咽癌CNE2细胞生长曲线的影响;CytationTM 5高通量活细胞成像分析检测系统分别监测3种不同浓度苯丙素类化合物对人鼻咽癌CNE2细胞形态改变;Western blot法检测3种苯丙素类化合物对人鼻咽癌CNE2细胞增殖相关蛋白增殖细胞核抗原（proliferating cell nuclear antigen， PCNA）、X连锁的凋亡蛋白抑制剂（X-linked inhibitor of apoptosis protein， XIAP）、Survivin表达的影响。结果 RTCA实时监测结果显示异欧前胡素、阿魏酸、绿原酸均能不同程度地抑制CNE2细胞增殖，存在药物浓度越高抑制效应越明显的趋势。CytationTM 5检测结果显示，与对照组相比，3种苯丙素类化合物作用36 h后，大量CNE2细胞出现细胞漂浮死亡，细胞膜破溃、细胞结构模糊等形态变化。Western blot检测结果显示，与对照组相比，异欧前胡素、阿魏酸、绿原酸能明显抑制CNE2细胞中增殖相关蛋白PCNA、XIAP、Survivin的表达（P<0.05）。结论 益气解毒方中主要苯丙素类化合物异欧前胡素、阿魏酸、绿原酸均可以不同程度抑制人鼻咽癌CNE2细胞增殖，其机制可能与下调增殖相关蛋白PCNA、XIAP、Survivin的表达有关。

〔关键词〕 鼻咽癌;益气解毒方;苯丙素类化合物;细胞增殖;异欧前胡素;阿魏酸;绿原酸

〔中图分类号〕R285.5       〔文献标志码〕A       〔文章编号〕doi：10.3969/j.issn.1674-070X.2021.05.009

Effects of the Phenylpropanoids in Yiqi Jiedu Formula on the Proliferation of

Nasopharyngeal Carcinoma Cells

XIONG Yu1， LIU Jie1， ZOU Pan1， SU Rui1， ZHONG Wenliang1， JIANG Zhichao2， WANG Xianwen3，4，5， HE Yingchun1，4，5*

（1. Hunan University of Chinese Medicine， Changsha， Hunan 410208， China; 2. Brain Hospital of Hunan Province， Changsha， Hunan 410005， China; 3. The First Affiliated Hospital of Hunan University of Chinese Medicine， Changsha， Hunan 410007， China; 4. Hunan Provincial Engineering and Technological Research Center for Prevention and Treatment of Ophthalmology and Otolaryngology Diseases with Chinese Medicine and Protecting Visual Function， Changsha， Hunan 410208， China; 5. Hunan Provincial Key Laboratory for the Prevention and Treatment of Ophthalmology and Otolaryngology Diseases with Chinese Medicine， Changsha， Hunan 410208， China）

〔Abstract〕 Objective To investigate the inhibitory effect and mechanism of the phenylpropanoid compounds isoimperatorin， ferulic acid and chlorogenic acid in Yiqi Jiedu Formula on the proliferation of human nasopharyngeal carcinoma CNE2 cells.  Methods Real-time unlabeled cell function analysis technique （RTCA） was used to dynamically monitor the effects of three phenylpropanoids at different concentrations on the proliferation of human nasopharyngeal carcinoma CNE2 cells. CytationTM 5 high-throughput live cell imaging analysis system was used to monitor the effects of three phenylpropanoids at different concentrations on the morphological changes of human nasopharyngeal carcinoma CNE2 cells. Western blot method was used to detect the effects of three phenylpropanoids on human nasopharyngeal carcinoma CNE2 cells proliferating cell nuclear antigen （PCNA）， X-linked inhibitor of apoptosis protein （XIAP）， and Survivin expression. Results RTCA results showed that isoperatorin， ferulic acid， and chlorogenic acid can inhibit the proliferation of CNE2 cells to varying degrees， and there was a trend that the higher the drug concentration， the more obvious the inhibitory effect. The CytationTM 5 results showed that compared with the control group， a large number of CNE2 cells had cell floating death， cell membrane rupture， cell structure fuzzy and other morphological changes after the three phenylpropanoids were treated for 36 hours. Western blot detection results showed that compared with the control group， isoperatorin， ferulic acid， and chlorogenic acid can significantly inhibit the expression of proliferation-related proteins PCNA， XIAP， and Survivin in CNE2 cells （P<0.05）. Conclusion The main phenylpropanoid compounds， isoimperatorin， ferulic acid and chlorogenic acid in Yiqi Jiedu Formula， which can inhibit the proliferation of human nasopharyngeal carcinoma CNE2 cells to varying degrees. The mechanism may be related to the down-regulation of proliferation-related proteins PCNA， XIAP， and Survivin expression level.

〔Keywords〕 nasopharyngeal carcinoma; Yiqi Jiedu Formula; phenylpropanoid compounds; cell proliferation; isoimperatorin; ferulic acid; chlorogenic acid

鼻咽癌（nasopharyngeal carcinoma， NPC）为发生在鼻咽上皮内壁的头颈部高发恶性肿瘤，发病率具有明显的地域倾向和种族差异，在中国南方、北非和东南亚发病率较高[1]。我国传统中医药在治疗NPC患者方面有独特优势，和单纯放疗相比，中西医结合疗法能有效改善NPC患者口腔黏膜不良反应、胃肠道不良反应及唾液腺损伤情况等，并能有效提高患者生活质量[2]。……