中国实验方剂学杂志2026，Vol.32Issue(19)：1-12,12.DOI:10.13422/j.cnki.syfjx.20251925
补中益气汤联合顺铂调控PDK1/Akt信号通路抑制糖酵解影响肺腺癌细胞增殖
Buzhong Yiqitang Combined with Cisplatin Inhibits Lung Adenocarcinoma Cell Proliferation by Suppressing PDK1/Akt Signaling Pathway and Regulating Glycolysis
摘要
Abstract
Objective:To investigate the impact of Buzhong Yiqitang combined with cisplatin on the proliferation of human lung adenocarcinoma(A549)cells through regulation of the pyruvate dehydrogenase kinase 1(PDK1)/protein kinase B(Akt)signaling pathway and influence on glycolysis.Methods:Transcriptome sequencing(RNA-seq)was employed to compare the expression of glycolysis-related genes between A549 cells and cisplatin-resistant human lung adenocarcinoma cells(A549/DDP).Small interfering RNA(siRNA)was employed to knock down PDK1,and the knockdown efficiency was verified by Western blot and Read-time PCR.The cell counting kit-8(CCK-8)assay was used to assess the survival and viability of A549 cells under the following conditions:siRNA negative control+cisplatin(128,64,32,16,8,4,0 μmol·L-1),siPDK1+cisplatin(128,64,32,16,8,4,0 μmol·L-1),and siPDK1+cisplatin(128,64,32,16,8,4,0 μmol·L-1)+Buzhong Yiqitang(10%)-containing serum.The 20%inhibitory concentration(IC20)of the siRNA negative control+cisplatin group(7.832 μmol·L-1)was calculated and used as the subsequent cisplatin concentration.Colony formation assay was performed to evaluate the proliferation of A549 cells.Lactate and adenosine triphosphate(ATP)assay kits were used to measure lactate and ATP production.The mitochondrial membrane potential was detected with the fluorescent probe JC-1.Western blotting was conducted to examine the expression levels of PDK1,phosphorylated(p)-Akt,Akt,pyruvate kinase M2(PKM2),glucose transporter 1(GLUT1),pyruvate dehydrogenase(PDH),and lactate dehydrogenase A(LDHA).Confocal immunofluorescence was employed to detect PDK1 and p-Akt.Results:RNA-seq results identified PDK1 as a highly expressed differential gene in glycolysis metabolism between A549 cells and A549/DDP cells,and it was highly expressed in tumor cells.Gene Set Enrichment Analysis(GSEA)revealed upregulated and downregulated genes in glycolysis and gluconeogenesis pathways.Western blot and RT-qPCR confirmed that PDK1-si-2 had the highest transfection efficiency,with a PDK1 knockdown rate exceeding 60%.CCK-8 assay determined the half-maximal inhibitory concentration(IC50)values for each group as(30.698±5.348),(16.372±3.562),(13.237±1.573)μmol·L-1,while the IC20 of cisplatin in siRNA negative control-transfected A549 cells was(7.832±0.672)μmol·L-1.Compared with the siRNA negative control group,the siRNA negative control+cisplatin group showed decreased colony formation rate,reduced lactate production,lowered mitochondrial membrane potential,downregulated protein levels of p-Akt,GLUT1,PDK1,PDH,and LDHA,and reduced PDK1 fluorescence intensity(P<0.05).The siPDK1 group exhibited decreased colony formation rate,reduced lactate production,increased ATP production,lowered mitochondrial membrane potential,downregulated protein levels of p-Akt,PKM2,GLUT1,PDK1,PDH,and LDHA,and reduced PDK1 and p-Akt fluorescence intensity(P<0.05).Compared with the siPDK1 group,the siPDK1+Buzhong Yiqitang group showed decreased colony formation rate,reduced lactate production,downregulated protein levels of PKM2,GLUT1,and PDK1,and reduced PDK1 fluorescence intensity(P<0.05).The siPDK1+cisplatin group exhibited decreased colony formation rate,reduced lactate production,increased ATP production,lowered mitochondrial membrane potential,downregulated protein levels of PKM2,GLUT1,PDK1,and PDH,and reduced PDK1 and p-Akt fluorescence intensity(P<0.05).The siPDK1+cisplatin+Buzhong Yiqitang group demonstrated decreased colony formation rate,reduced lactate production,increased ATP production,lowered mitochondrial membrane potential,downregulated protein levels of p-Akt,PKM2,GLUT1,PDK1,PDH,and LDHA,and reduced PDK1 and p-Akt fluorescence intensity(P<0.05).Compared with the siPDK1+Buzhong Yiqitang group,the siPDK1+cisplatin group showed decreased colony formation rate,downregulated protein levels of p-Akt and PDH,and reduced PDK1 and p-Akt fluorescence intensity(P<0.05).The siPDK1+cisplatin+Buzhong Yiqitang group exhibited decreased colony formation rate,reduced lactate production,increased ATP production,lowered mitochondrial membrane potential,downregulated protein levels of p-Akt,GLUT1,PDH,and LDHA,and reduced PDK1 and p-Akt fluorescence intensity(P<0.05).Compared with the siPDK1+cisplatin group,the siPDK1+cisplatin+Buzhong Yiqitang group showed decreased colony formation rate,increased ATP production,and downregulated protein levels of p-Akt,PKM2,and LDHA(P<0.05).Conclusion:Buzhong Yiqitang combined with cisplatin can suppress lung adenocarcinoma cell proliferation by modulating glycolysis through the PDK1/Akt signaling pathway.关键词
补中益气汤/顺铂/糖酵解/丙酮酸脱氢酶激酶1(PDK1)/蛋白激酶B(Akt)/小干扰RNA(siRNA)Key words
Buzhong Yiqitang/cisplatin/glycolysis/pyruvate dehydrogenase kinase 1(PDK1)/protein kinase B(Akt)/small interfering RNA(siRNA)分类
医药卫生引用本文复制引用
李贺,白思嘉,刘文俊,王建光,吕佳璐,王淳..补中益气汤联合顺铂调控PDK1/Akt信号通路抑制糖酵解影响肺腺癌细胞增殖[J].中国实验方剂学杂志,2026,32(19):1-12,12.基金项目
国家自然科学基金项目(82174254) （82174254）
辽宁中医药大学中医药科技创新多学科交叉团队项目(2025-Dxkjc-15-02) （2025-Dxkjc-15-02）
辽宁省教育厅科技创新团队项目(LJ222510162018) （LJ222510162018）
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) （2023QJ01001）
辽宁中医药大学"中医脏象理论及应用教育部重点实验室"2022年度开放基金项目(zyzx2204) （zyzx2204）
