文献类型：期刊文献

中文题名：敦煌方大补肺汤联合顺铂对Lewis肺腺癌荷瘤小鼠的干预作用及分子机制

英文题名：Intervention Effect and Molecular Mechanism of Dabufei Decoction in Dunhuang Formula Combined with Cisplatin on Lewis Lung Adenocarcinoma in Mice

作者：肖孟勇[1];骆亚莉[1,2];齐晓风[1];罗晶[1];马琳娜[1];阮琳峰[1];连妮妮[1]

第一作者：肖孟勇

机构：[1]甘肃中医药大学,兰州730000;[2]甘肃省中医药防治慢性疾病重点实验室,兰州730000

第一机构：甘肃中医药大学

年份：2024

卷号：30

期号：17

起止页码：95

中文期刊名：中国实验方剂学杂志

外文期刊名：Chinese Journal of Experimental Traditional Medical Formulae

收录：CSTPCD;;Scopus;北大核心:【北大核心2023】；CSCD:【CSCD2023_2024】；

基金：国家自然科学基金项目(82160846);甘肃省“双一流”科研重点项目(GSSYLXM-05)。

语种：中文

中文关键词：大补肺汤;顺铂;Lewis肺腺癌;免疫细胞;磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)/哺乳动物雷帕霉素靶蛋白(mTOR)信号通路

外文关键词：Dabufei decoction;cisplatin;Lewis lung adenocarcinoma;immune cell;phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)/mammalian target of rapamycin(mTOR)signaling pathway

摘要：目的:探讨敦煌方大补肺汤联合顺铂对Lewis肺腺癌荷瘤小鼠的干预作用及相关分子机制。方法:C57BL/6J小鼠50只,随机挑选10只作为空白组(不造模),40只皮下接种Lewis细胞,复制Lewis肺腺癌荷瘤小鼠模型,随机分为模型组(等量生理盐水)、顺铂组(5 mg·kg^(-1))、大补肺汤组(14.35 g·kg^(-1)·d^(-1))、大补肺汤联合顺铂组(大补肺汤14.35 g·kg^(-1)·d^(-1)+顺铂5 mg·kg^(-1)),每组10只,连续治疗14 d。观察小鼠一般状况,记录体质量变化,计算抑瘤率、脾指数、胸腺指数,血常规检测外周血白细胞(WBC)、血小板(PLT)、血红蛋白(HGB),流式细胞术检测脾脏CD4^(+)CD25^(+)FoxP3^(+)调节性T细胞(Treg)、自然杀伤(NK)细胞表达,蛋白免疫印迹法(Western blot)及实时荧光定量聚合酶链式反应(Real-time PCR)测定肿瘤组织内磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)/哺乳动物雷帕霉素靶蛋白(mTOR)信号通路相关蛋白及mRNA的表达。结果:与空白组比较,模型组体质量降低(P<0.05),脾指数、胸腺指数明显降低(P<0.05),脾脏中NK细胞百分比明显下降(P<0.05),Treg细胞百分比明显升高(P<0.05),WBC、PLT、HGB数量均明显下降(P<0.05);与模型组比较,大补肺汤组肿瘤生长明显抑制,体质量升高,瘤质量明显降低(P<0.05),NK细胞百分比明显升高(P<0.05),Treg细胞比例明显降低(P<0.05),WBC、PLT、HGB数量明显升高(P<0.05),顺铂组肿瘤生长明显抑制,体质量明显降低(P<0.05),瘤质量明显降低(P<0.05),胸腺指数、脾指数明显降低(P<0.05),Treg细胞百分比明显降低(P<0.05),WBC、PLT、HGB数量明显下降(P<0.05),大补肺汤联合顺铂组肿瘤生长明显抑制,瘤质量明显降低(P<0.05),各给药组肿瘤组织中PI3K、Akt、mTOR蛋白磷酸化及mRNA水平明显降低(P<0.05);与顺铂组比较,大补肺汤联合顺铂组肿瘤生长明显抑制,瘤质量明显降低(P<0.05),体质量明显升高(P<0.05),脾指数、胸腺指数明显升高(P<0.05),NK细胞明显升高(P<0.05),Treg细胞百分比明显降低(P<0.05),WBC、PLT、HGB数量明显增多(P<0.05),PI3K、Akt、mTOR磷酸化及mRNA水平明显降低(P<0.05)。结论:大补肺汤联合顺铂具有增效减毒作用,可有效调节免疫功能,增加NK细胞比例,降低Treg细胞比例,改善骨髓抑制,下调PI3K/Akt/mTOR信号通路抑制Lewis肺腺癌荷瘤小鼠的肿瘤生长。
Objective:To explore the intervention effect and molecular mechanism of Dabufei decoction in Dunhuang formula combined with cisplatin on Lewis lung adenocarcinoma-bearing mice.Method:Fifty C57BL/6J mice were used,with 10 randomly assigned to the blank group(without modeling),and 40 subcutaneously inoculated with Lewis cells to establish a Lewis lung adenocarcinoma-bearing mouse model.These 40 mice were randomly divided into the following four groups(with 10 mice in each group):Model group(equal volume of physiological saline),cisplatin group(5 mg·kg^(-1)),Dabufei decoction group(14.35 g·kg^(-1)·d^(-1)),and Dabufei decoction combined with cisplatin group(Dabufei decoction 14.35 g·kg^(-1)·d^(-1)+cisplatin 5 mg·kg^(-1)).Each group was treated continuously for 14 days.The general condition of the mice was observed,body weight changes were recorded,and the tumor inhibition rate,spleen index,and thymus index were calculated.Peripheral blood white blood cell(WBC),platelet(PLT),and hemoglobin(HGB)were detected by routine blood tests.Flow cytometry was used to detect the expression of CD4^(+)CD25^(+)FoxP3^(+)regulatory T cells(Treg)and natural killer(NK)cells in the spleen.Western blot and real-time quantitative polymerase chain reaction(Real-time PCR)were used to determine the expression of proteins and mRNA related to the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)/mammalian target of rapamycin(mTOR)signaling pathway in tumor tissues.Result:Compared with the blank group,the model group showed decreased body weight(P<0.05),spleen index,and thymus index(P<0.05),decreased percentage of NK cells in the spleen(P<0.05),increased percentage of Treg cells(P<0.05),and decreased counts of WBC,PLT,and HGB(P<0.05).Compared with the model group,the Dabufei decoction group exhibited significant tumor growth inhibition,increased body weight,and reduced tumor weight(P<0.05),increased percentage of NK cells(P<0.05),decreased proportion of Treg cells(P<0.05),and increased counts of WBC,PLT,and HGB(P<0.05).In the cisplatin group,tumor growth was significantly inhibited,body weight significantly decreased(P<0.05),and tumor weight significantly reduced(P<0.05).The spleen index and thymus index decreased(P<0.05),and the percentage of Treg cells significantly decreased(P<0.05).The counts of WBC,PLT,and HGB significantly decreased(P<0.05).In the Dabufei decoction combined with cisplatin group,tumor growth was significantly inhibited,and tumor weight significantly reduced(P<0.05).The levels of phosphorylated PI3K,Akt,and mTOR proteins and mRNA in tumor tissues were significantly reduced in all medication groups(P<0.05).Compared with the cisplatin group,the Dabufei decoction combined with cisplatin group showed significantly inhibited tumor growth,reduced tumor weight(P<0.05),increased body weight(P<0.05),increased spleen index and thymus index(P<0.05),increased percentage of NK cells(P<0.05),decreased percentage of Treg cells(P<0.05),significantly increased counts of WBC,PLT,and HGB(P<0.05),and reduced levels of phosphorylated PI3K,Akt,and mTOR and their mRNA(P<0.05).Conclusion:Dabufei decoction combined with cisplatin has a synergistic effect with reduced toxicity,effectively regulating immune function,increasing the proportion of NK cells,reducing the proportion of Treg cells,improving bone marrow suppression,and downregulating the PI3K/Akt/mTOR signaling pathway to inhibit tumor growth in Lewis lung adenocarcinoma-bearing mice.