补中益气汤通过Nrf2/ROS通路调控内质网应激改善A549细胞、A549/DDP细胞顺铂耐药的分子机制  

Buzhong Yiqi Decoction(补中益气汤)Regulates Endoplasmic Reticulum Stress through Nrf2/ROS Pathway to Improve Molecular Mechanism of Cisplatin Resistance in A549 Cells and A549/DDP Cells

作　　者：于丹[1] 牟琪瑞 高原[1] YU Dan;MOU Qirui;GAO Yuan(Department of Pathology,Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China)

机构地区：[1]辽宁中医药大学病理学教研室,辽宁沈阳110847

出　　处：《中华中医药学刊》2025年第3期170-174,I0034,共6页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81973735);辽宁省自然科学基金项目(2023-MS-232);辽宁中医药大学中西医结合学院“岐济”人才支撑计划项目(2023QJ01001)。

摘　　要：目的通过对补中益气汤调控Nrf2/ROS通路对A549细胞、A549/DDP细胞内质网应激的影响,探讨补中益气汤改善非小细胞肺癌顺铂耐药的分子机制。方法制备补中益气汤含药血清并培养A549细胞、A549/DDP细胞,并进行随机分组为A组(A549细胞)、B组(A549细胞+20μg/mL顺铂)、C组(A549细胞+20μg/mL顺铂+10%补中益气汤含药血清)、D组(A549/DDP细胞)、E组(A549/DDP细胞+20μg/mL顺铂)、F组(A549/DDP细胞+20μg/mL顺铂+10%补中益气汤含药血清),应用CCK-8法检测各组细胞顺铂半数抑制浓度(IC50)值,DCFH-DA荧光探针法检测各组活性氧(ROS)含量,Western blot法检测各组细胞Nrf2、P-Nrf2、PERK、P-PERK、eIF2α、P-eIF2α、IRE1、XBP1的蛋白表达量。结果在A549细胞和A549/DDP细胞中,补中益气汤均可以显著下调其对顺铂的IC50值(P<0.05);两株细胞中,顺铂均可以上调Nrf2、P-Nrf2的蛋白表达,在补中益气汤干预后,二者表达量下调(P<0.05),且二者在A549/DDP细胞中表达较A549细胞更高(P<0.05);与空白组比较,顺铂可以上调两株细胞的ROS表达量(P<0.05),在联合了补中益气汤后ROS表达量进一步上调(P<0.05),且A549细胞的ROS表达量要显著高于A549/DDP细胞(P<0.05);PERK、eIF2α在各组细胞中表达无显著差异(P>0.05),在两株细胞中,顺铂能显著上调P-PERK、P-eIF2α、IRE1、XBP1、蛋白表达,在补中益气汤干预后表达再次上调(P<0.05),且A549细胞的内质网应激蛋白表达高于A549/DDP细胞(P<0.05)。结论补中益气汤可能通过Nrf2/ROS通路调控内质网应激改善A549细胞、A549/DDP细胞顺铂耐药。Objective By studying the effect of Buzhong Yiqi Decoction(补中益气汤)on the endoplasmic reticulum stress of A549 cells and A549/DDP cells through Nrf2/ROS pathway,the molecular mechanism of Buzhong Yiqi Decoction improving cis-platin resistance in non-small cell lung cancer was investigated.Method The serum containing Buzhong Yiqi Decoction was pre-pared and A549 cells and A549/DDP cells were cultured.They were randomly divided into group A(A549 cells),Group B(A549 cells+20μg/mL cisplatin),group C(A549 cells+20μg/mL cisplatin+10%Buzhong Yiqi Decoction-containing ser-um),group D(A549/DDP cells),group E(A549/DDP cells+20μg/mL cisplatin)and group F(A549/DDP cells+20μg/mL cisplatin+10%Buzhong Yiqi Decoction-containing serum).Half inhibitory concentration of cisplatin(IC50)was detected by CCK-8 method,and reactive oxygen species(ROS)were detected by DCFH-DA fluorescent probe method.The protein expres-sion levels of Nrf2,P-Nrf2,PERK,P-PERK,eIF2α,P-eIF2α,IRE1 and XBP1 were detected by Western blot.Result In both A549 cells and A549/DDP cells,Buzhong Yiqi Decoction significantly decreased the IC50 value of cisplatin(P<0.05).In both cells,cisplatin could up-regulate the protein expressions of Nrf2 and P-Nrf2,and after the intervention of Buzhong Yiqi Decoc-tion,the expression levels of both were down-regulated(P<0.05)in A549/DDP cells compared with those in A549 cells(P<0.05).Compared with the blank group,cisplatin could up-regulate the ROS expression of the two cells(P<0.05),the ROS expression was further up-regulated after combined with Buzhong Yiqi Decoction(P<0.05),and the ROS expression level of A549 cells was significantly higher than that of A549/DDP cells(P<0.05).There were no significant differences in the expres-sions of PERK and eIF2αamong all groups(P>0.05).Cisplatin significantly up-regulated the protein expressions of P-PERK,P-eIF2α,IRE1 and XBP1 in the two cell lines,and the expression was up-regulated again after the intervention of Buzhong Yiqi Decoction(P<0.05),and the expression of ER stre

关 键 词：补中益气汤 非小细胞肺癌 Nrf2/ROS 内质网应激 顺铂耐药 

分 类 号：R289.5[医药卫生—方剂学]