基于PI3K/AKT信号通路探讨清肺消积颗粒对Lewis肺癌小鼠肿瘤细胞凋亡的影响  

Effect of Qingfeixiaoji granules on apoptosis of Lewis lung cancer mice based on PI3K/AKT pathway

作　　者：张格松 程建通 张灏[1] 张蕊 沈晓倩 赵艺涵 蒋士卿[2] ZHANG Ge-song;CHENG Jian-tong;ZHANG Hao;ZHANG Rui;SHEN Xiao-qian;ZHAO Yi-han;JIANG Shi-qing(Taicang Hospital of Traditional Chinese Medicine Geriatrics Department,Suzhou 215499,China;The First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450099,China)

机构地区：[1]太仓市中医医院老年病科,苏州215499 [2]河南中医药大学第一附属医院血液肿瘤科,郑州450099

出　　处：《天然产物研究与开发》2025年第2期215-222,共8页Natural Product Research and Development

基　　金：河南省自然科学基金(232300420055);张仲景传承与创新专项(GZY-KJS-2022-038-2);河南省中医药文化与管理研究项目(TCM2022002)。

摘　　要：研究清肺消积颗粒(Qingfeixiaoji granules,QFXJ)对Lewis肺癌小鼠肿瘤生长的抑制作用及对肿瘤细胞凋亡的影响,选取45只雄性C57BL/6J小鼠,建立Lewis肺癌小鼠模型,随机分为模型组(model group,Mod)、顺铂组(cisplatin group,DDP)、QFXJ低剂量组(low dose QFXJ group,QFXJ-L)、QFXJ中剂量组(middle dose QFXJ group,QFXJ-M)、QFXJ高剂量组(high dose QFXJ group,QFXJ-H),每组9只,另选9只C57BL/6J小鼠设为空白组(control group,Con)。DDP腹腔注射0.2 mL顺铂(剂量为2 mg/kg),2 d/次,QFXJ-L、QFXJ-M、QFXJ-H灌胃0.4 mL QFXJ(剂量依次为12、24、48 g/kg),1 d/次,连续给药14 d后剥离肿瘤组织并收集血液,计算抑瘤率;苏木精-伊红染色法(hematoxylin-eosin,HE)观察肿瘤组织病理情况;酶联免疫吸附(enzyme-linked immunosorbent assay,ELISA)法测定血清中肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-6(interleukin-6,IL-6)和白细胞介素-1β(interleukin-1β,IL-1β)的含量;蛋白质免疫印迹(Western blot,WB)法测定肿瘤组织中磷脂酰肌3-激酶(phosphatidylinositol 3 kinase,PI3K)、磷酸化蛋白激酶B(phosphorylated protein kinase B,p-AKT)、B细胞淋巴瘤-2(B cell lymphocyte tumor 2,Bcl-2)、B细胞淋巴瘤-2相关X蛋白(Bcl-2 associated X protein,Bax)、半胱天冬酶3(cysteinyl aspartate specific proteinase-3,Caspase-3)的表达水平;原位末端转移酶标记(TdT-mediated dUTP nick-end labeling,TUNEL)技术观察肿瘤组织中细胞的凋亡情况。结果发现,与Mod相比,QFXJ-M、QFXJ-H与DDP瘤重均显著降低(P<0.05);肿瘤组织病理结果显示,QFXJ-L、QFXJ-M、QFXJ-H肿瘤细胞密度均有不同程度降低,坏死细胞密度增加;QFXJ-M、QFXJ-H与DDP血清中TNF-α、IL-6和IL-1β表达量均显著降低(P<0.05);QFXJ-H与DDP肿瘤组织中PI3K、p-AKT、Bcl-2蛋白表达水平降低,Bax、Caspase-3蛋白表达水平显著升高(P<0.05);TUNEL荧光结果显示,各给药组阳性细胞数量较模型组均有不同程度增多。得出结论,QFXJ可能通过调控PTo study the inhibition effects of Qingfeixiaoji granules(QFXJ)on Lewis lung cancer mice and the apoptosis of tumor cells,45 male C57BL/6J mice were selected to build the Lewis lung cancer mouse model.They were randomly divided into model group(Mod),cisplatin group(DDP),low dose QFXJ group(QFXJ-L),middle dose QFXJ group(QFXJ-M),and high dose QFXJ group(QFXJ-H),with nine mice in each group.Other nine C57BL/6J mice were selected as the control group(Con).DDP was intraperitoneally injected with 0.2 mL cisplatin(according to 2 mg/kg)once every two days,and QFXJ-L,QFXJ-M,and QFXJ-H groups were intragastrically injected with 0.4 mL QFXJ in sequence(according to 12,24,and 48 g/kg,respectively)once a day.After 14 days of continuous administration,the tumor inhibition rate was calculated by stripping the tumor tissues and collecting blood.Hematoxylin-eosin(HE)staining was used to observe the pathological status of tumor tissues.Enzyme-linked immunosorbent assay(ELISA)was applied to determine the contents of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and interleukin-1β(IL-1β).The expression levels of phosphatidylinositol 3 kinase(PI3K),phosphorylated protein kinase B(p-AKT),B cell lymphocyte tumor 2(Bcl-2),Bcl-2 associated X protein(Bax),and cysteinyl aspartate specific proteinase-3(Caspase-3)in tumor tissues were determined with Western blot(WB).Cell apoptosis in tumor tissues was observed with TdT-mediated dUTP nick-end labeling(TUNEL)technology.The results showed that the tumor weight of QFXJ-M,QFXJ-H,and DDP was significantly decreased compared with Mod(P<0.05).The tumor histopathological results suggested that the density of tumor cells in QFXJ-L,QFXJ-M,and QFXJ-H decreased to varying degrees,and the density of necrotic cells increased.The expressions of TNF-α,IL-6,and IL-1βin the serum of QFXJ-M,QFXJ-H,and DDP were significantly decreased(P<0.05).The protein expression levels of PI3K,p-AKT,and Bcl-2 were decreased in tumor tissues of QFXJ-H and DDP,while the expression levels of Bax and Caspase-3 were s

关 键 词：清肺消积颗粒 肺癌 PI3K/AKT通路 凋亡 

分 类 号：R273[医药卫生—中西医结合]