基于PI3K/Akt信号通路探讨姜黄素诱导A549细胞凋亡的机制  

Mechanism of curcumin induced apoptosis of A549 cells based on PI3K/Akt signal pathyvay

作　　者：刘嘉欣[1] 黎同明[2] 黄慧贤 陈少娜 陈学增 LIU Jia-xin;LI Tong-ming;HUANG Hui-xian;CHEN Shao-na;CHEN Xue-zeng(Pharmaceutical Department,The First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine,Guangzhou 510405,Guangdong Province,China;College of Traditional Chinese Medicine,Guangzhou University of Traditional Chinese Medicine,Guangzhou 510006,Guangdong Province,China)

机构地区：[1]广州中医药大学第一附属医院药学部,广东广州510405 [2]广州中医药大学中药学院,广东广州510006

出　　处：《中国临床药理学杂志》2023年第17期2487-2491,共5页The Chinese Journal of Clinical Pharmacology

基　　金：广东省中医药管理局面上基金资助项目(20221152)。

摘　　要：目的 观察并探讨姜黄素对非小细胞肺癌A549细胞的作用及其机制。方法 将A549细胞分为空白组(正常培养)和低、高剂量实验组(分别用40、80μmol·L^(-1)姜黄素干预48 h)及激动药+低、高剂量实验组(先用50μmol·L^(-1)Recilisib Sodium干预24 h后,再分别用40、80μmol·L^(-1)姜黄素干预48 h)、抑制药+低、高剂量实验组(先用25μmol·L^(-1)PI3K-IN^(-1)干预24 h后,再分别用40、80μmol·L^(-1)姜黄素干预48 h)。用MTT法测定细胞增值率;用流式细胞术测定细胞凋亡情况;用实时荧光聚合酶链反应(qRT-PCR)检测磷脂酰肌醇3激酶(PI3K)、蛋白质丝氨酸苏氨酸激酶(Akt)mRNA的相对表达水平;用蛋白质印迹法检测PI3K、p-PI3K、Akt、p-Akt蛋白表达水平。结果 空白组、低剂量实验组、高剂量实验组、激动药+低剂量实验组、抑制药+低剂量实验组、激动药+高剂量实验组、抑制药+高剂量实验组的细胞活力分别为(100.00±6.59)%、(94.99±4.90)%、(93.72±3.41)%、(88.32±2.34)%、(66.88±4.10)%、(43.98±7.49)%和(26.99±2.64)%;细胞凋亡率分别为(4.96±0.65)%、(24.28±1.36)%、(34.79±3.94)%、(11.10±2.41)%、(14.07±1.52)%、(35.71±3.51)%和(45.87±3.32)%;PI3K mRNA表达水平分别为1.00±0.00、0.69±0.04、0.49±0.06、0.81±0.11、0.52±0.04、0.69±0.03和0.25±0.03;Akt mRNA表达水平分别为1.00±0.00、0.68±0.06、0.43±0.11、0.82±0.09、0.50±0.10、0.62±0.08和0.31±0.07;p-PI3K/PI3K比值分别为1.00±0.13、0.58±0.16、0.45±0.22、1.23±0.49、0.75±0.30、1.03±0.23和0.33±0.24;p-Akt/Akt比值分别为1.00±0.13、0.67±0.07、0.44±0.14、0.81±0.05、0.42±0.09、0.71±0.04和0.22±0.05。上述指标,低剂量实验组、高剂量实验组与空白组比较,激动药+低剂量实验组、抑制药+低剂量实验组与低剂量实验组比较,激动药+高剂量实验组、抑制药+高剂量实验组与高剂量实验组比较,差异均有统计学意义(P<0.05,P<0.01)。结论 姜黄素对非小细胞肺癌A549�Objective To observe and explore the effect and mechanism of curcumin on non-small cell lung cancer A549 cells.Method1A549 cellsweredivided into blank group(cultured under normoxic conditions),experimental-L,-H groups(40,80μmol·L^(-1) curcumin culture for 48 h),activator+experimental-L group(Recinib Society was cultured for 24 h,and 40μmol·L^(-1)curcumin was cultured for 48 h),inhibitor+experimental-Lgroup(PI3K-IN-1 culture for 24 h,and 40μmol-L^(-1) curcumin was cultured for 48 h),activator+experimental-H group(Recinib Society was cultured for 24 h,and 80μmol·L^(-1)curcumin for 48 h),inhibitor+experimental-H group(PI3K-IN-1 was cultured for 24 h,and 80μmol·L^(-1)curcumin was cultured for 48 h).MTT method was used to determine the effect of curcumin on cell proliferation rate;cell apoptosis was measured by flow cytometry;quantitative real time polymerase chain reaction was used to detect the mRNA expression levels of phosphatidylinositol 3 kinase(PI3K)and protein serine threonine kinase(Akt)mRNA;Western blotting was used to detect the expressions of PI3K,p-PI3K,Akt,and p-Akt proteins.Result The cell viabilites of blank,experimental-L,-H groups,activator+experimental-L group,inhibitor+experimental-L group,activator+experimental-H group,inhibitor+experimental-H group were(100.00±6.59)%,(94.99±4.90)%,(93.72±3.41)%,(88.32±2.34)%,(66.88±4.10)%,(43.98±7.49)%and(26.99±2.64)%,respectively;the apoptosis rates were(4.96±0.65)%,(24.28±1.36)%,(34.79±3.94)%,(11.10±2.41)%,(14.07±1.52)%,(35.71±3.51)%and(45.87±3.32)%,respectively;the expression levels of PI3K mRNA were 1.00±0.00,0.69±0.04,0.49±0.06,0.81±0.11,0.52±0.04,0.69±0.03 and 0.25±0.03,respectively;the Akt mRNA expression levels were 1.00±0.00,0.68±0.06,0.43±0.11,0.82±0.09,0.50±0.10,0.62±0.08 and 0.31±0.07,respectively;the p-PI3K/PI3K ratios were 1.00±0.13,0.58±0.16,0.45±0.22,1.23±0.49,0.75±0.30,1.03±0.23 and 0.33±0.24,respectively;the p-Akt/Akt ratios were 1.00±0.13,0.67±0.07,0.44±0.14,0.81±0.05,0.42±0.09,0.71±0.04 and 0.2

关 键 词：姜黄素 肺癌 A549细胞 磷脂酰肌醇3激酶/蛋白质丝氨酸苏氨酸激酶信号通路 

分 类 号：R97[医药卫生—药品]