11-羰基-β-乙酰乳香酸抑制口腔鳞状细胞癌的研究  

Study of acetyl-11-keto-3-boswellic acid inhibiting oral squamous cell carcinoma

作　　者：黄新帮[1] 王羽[1] 俞倩 李勇[1] 李标东 何海蕾[1] 陈琨[1] 赵玲帆 HUANG Xin-bang;WANG Yu;YU Qian;LI Yong;LI Biao-dong;HE Hai-lei;CHEN Kun;ZHAO Ling-fan(Oral and Maxillofacial Surgery,Ganzhou People's Hospital,Ganzhou 341000,Jiangxi Province,China;Department of Anesthesiology,Ganzhou People's Hospital,Ganzhou 341000,Jiangxi Province,China)

机构地区：[1]赣州市人民医院口腔颌面外科,江西赣州341000 [2]赣州市人民医院麻醉科,江西赣州341000

出　　处：《中国临床药理学杂志》2024年第20期2993-2997,共5页The Chinese Journal of Clinical Pharmacology

基　　金：江西省卫生健康委科技计划基金资助项目(SKJP220236690)。

摘　　要：目的探讨11-羰基-β-乙酰乳香酸(AKBA)诱导口腔鳞状细胞癌(OSCC)细胞凋亡的作用机制。方法将人舌鳞癌细胞CAL27随机分为对照组(常规培养)、低剂量组(40.00μmol·L^(-1)AKBA)、中剂量组(80.00μmol·L^(-1)AKBA)、高剂量组(120.00μmol·L^(-1)AKBA)、3-MA组[120.00μmol·L^(-1)AKBA+2 mmol·L^(-1)自噬抑制药3-甲基腺嘌呤(3-MA)]。用5-乙炔基-2’-脱氧尿苷(Edu)实验检测细胞增殖情况,用蛋白质印迹法检测自噬和凋亡相关蛋白表达水平,用流式细胞术检测细胞凋亡情况。将小鼠随机分为模型组(构建OSCC小鼠模型)、AKBA-L组(建模后灌胃10.00 mg·kg^(-1)AKBA)、AKBA-H组(建模后灌胃20.00 mg·kg^(-1)AKBA),每组10只。连续给药28 d后检测肿瘤质量,用蛋白质印迹法检测相关蛋白相对表达水平。结果对照组和高剂量组Edu阳性细胞率分别为(40.18±2.53)%和(12.08±0.93)%;对照组、高剂量组和3-MA组细胞自噬相关的微管相关蛋白1轻链3(LC3)Ⅱ/LC3Ⅰ蛋白比值分别为0.33±0.05、2.93±0.39和0.56±0.07,磷酸化的腺苷酸活化蛋白激酶催化亚基-α亚基(p-PRKAA1)蛋白相对表达水平分别为0.34±0.04、1.03±0.07和0.99±0.09,细胞凋亡率分别为(4.65±0.39)%、(25.75±2.29)%和(14.92±1.49)%。高剂量组的上述指标与对照组比较,在统计学上差异均有统计学意义(均P<0.05),3-MA组的上述指标与高剂量组比较,在统计学上差异均有统计学意义(均P<0.05)。模型组、AKBA-L组、AKBA-H组肿瘤质量分别为(0.96±0.08)、(0.55±0.06)和(0.43±0.05)g,LC3Ⅱ/LC3Ⅰ蛋白比值分别为0.47±0.09、0.94±0.21和1.69±0.34。AKBA-L组、AKBA-H组的上述指标与模型组比较,在统计学上差异均有统计学意义(均P<0.05)。结论AKBA可诱导细胞毒性自噬相关凋亡,抑制CAL27细胞增殖,这可能与激活AMPK信号相关。Objective To investigate the mechanism of apoptosis induced by acetyl-11-keto-3-boswellic acid(AKBA)in oral squamous cell carcinoma(OSCC)cells.Methods CAL27 were randomly divided into control group(conventional culture),low-dose group(40.00μmol·L^(-1)AKBA),middle-dose group(80.00μmol·L^(-1)AKBA),high-dose group(120.00μmol·L^(-1)AKBA),3-methyladenine(3-MA)group(120.00μmol·L^(-1)AKBA+2 mmol·L^(-1)autophagy inhibitor 3-MA).5-ethynyl-2'-deoxyuridine(Edu)assay was used to detect cell proliferation;Western blot assay was used to detect protein expression;flow cytometry was used to detect apoptosis.Mice were randomly divided into model group(construct OSCC mouse model),AKBA-L group(10.00 mg·kg^(-1)AKBA after modeling),AKBA-H group(20.00 mg·kg^(-1)AKBA after modeling),10 animals per group.After 28 days of continuous administration,weight were detected;and the expression of related proteins were detected by Western blot assay.Results The Edu positive cell rates in control group,high-dose group were(40.18±2.53)%,(12.08±0.93)%,respectively;the protein levels of autophagy associated microtubule associated protein 1 light chain 3(LC3)Ⅱ/LC3Ⅰin control group,high—dose group and 3-MA group were 0.33±0.05,2.93±0.39,0.56±0.07,respectively;phosphorylated adenylate activated protein kinase catalytic subunit alpha subunit 1(p-PRKAA1)protein levels were 0.34±0.04,1.03±0.07,0.99±0.09,respectively;the apoptosis rates were(4.65±0.39)%,(25.75±2.29)%,(14.92±1.49)%,respectively.The above indexes in hige-dose group were significantly different from those in the control group(all P<0.05).The above indexes in 3-MA group were significantly different from those in high-dose group(all P<0.05).The tumor weight of model group,AKBA-L group and AKBA-H group were(0.96±0.08),(0.55±0.06),(0.43±0.05)g,respectively;the protein levels of LC3Ⅱ/LC3Ⅰwere 0.47±0.09,0.94±0.21and 1.69±0.34,respectively.The above indexes in AKBA-L group and AKBA-H group were significantly different from those in model group(all P<0.05).Conclu

关 键 词：11-羰基-Β-乙酰乳香酸 口腔鳞状细胞癌 自噬 凋亡 腺苷酸活化蛋白激酶信号通路 

分 类 号：R28[医药卫生—中药学]