西黄丸组分中药调节肿瘤微环境中Treg细胞PI3K/AKT通路的抗肿瘤作用机制研究
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摘要
目的研究西黄丸组分中药是否与西黄丸具有相似的抗癌作用及机制。方法 4T1乳腺癌细胞荷瘤建立动物模型,西黄丸低、中、高剂量(0.39、0.78、1.95 g/kg)和西黄丸组分中药0.3 mL(74.5 mg/kg榄香烯、2.73 mg/kg牛磺酸、0.52mg/kg麝香酮和4.75 mg/kg 11-羰基-β乙酯乳香酸混合物,对应于西黄丸高剂量组)ig给药14 d,剥离肿瘤组织,称质量,匀浆,制备单细胞悬液,免疫磁硃分离Treg细胞。流式细胞术和免疫组化检测肿瘤微环境中Treg细胞数量变化情况,Western Blotting检测肿瘤微环境中Treg细胞磷脂酰肌醇3-激酶/蛋白激酶B (PI3K/AKT)蛋白表达情况,实时荧光定量PCR(qRT-PCR)检测肿瘤微环境中Treg细胞PI3K、AKT mRNA表达情况。结果与模型组及西黄丸低、中剂量组比较,西黄丸组分中药组肿瘤质量显著减少(P<0.05);流式细胞术和免疫组化结果显示,肿瘤微环境中Treg细胞数量显著减少(P<0.05);Western Blotting和qRT-PCR结果显示,肿瘤微环境中Treg细胞PI3K、AKT蛋白及mRNA表达显著下降(P<0.05)。与西黄丸高剂量组比较,西黄丸组分中药组肿瘤质量、Treg细胞数量和PI3K、AKT蛋白及mRNA表达均明显升高(P<0.05)。结论西黄丸组分中药通过抑制肿瘤微环境中Treg细胞PI3K/AKT信号通路的表达减少Treg细胞数量,进而表现与西黄丸相似的抗癌作用和机制。
Objective To study whether the Xihuang Pills component-based Chinese medicine has similar anti-cancer effect and mechanism as Xihuang Pills. Methods 4 T1 breast cancer cells were used to establish a tumor model in mice. The model mice were ig administered with Xihuang Pills of low, medium, and high doses (0.39, 0.78 and 1.95 g/kg) and Xihuang Pills component-based Chinese medicine of 0.3 mL (74.5 mg/kg elemene, 2.73 mg/kg taurine, 0.52 mg/kg Muscone and 4.75 mg/kg 11-carbonyl-beta-ethyl ester mastic acid mixture, corresponding to the high dosage group of Xihuang Pills) for 14 d. The tumor tissues were removed,weighed and homogenized to prepare single cell suspensions. Treg cells were prepared by magnetic cell sorting. The changes in number of Treg cells in the tumor microenvironment were detected by flow cytometry and immunohistochemistry. The protein expression of PI3K and AKT in Treg cells in the tumor microenvironment was detected by western blotting. The mRNA expression of PI3 K and AKT in Treg cells in the tumor microenvironment was detected by the Real-time PCR (qRT-PCR). Results Compared with model group and Xihuang Pills low-and medium-dose groups, the weight of tumors was significantly decreased in the Xihuang Pills component-based Chinese medicine group (P < 0.05); The results of flow cytometry and immunohistochemistry showed a significant decrease in the number of Treg cells in the tumor microenvironment (P < 0.05); Western Blotting and qRT-PCR results showed that the expression of PI3 K, AKT protein and their mRNA in Treg cells in the tumor microenvironment was significantly decreased (P < 0.05). Compared with the Xihuang Pills high-dose group, the tumor weight, the number of Treg cells, and the protein expression of PI3 K and AKT and their mRNA expression in the Xihuang Pills component-based Chinese medicine group were significant increased (P < 0.05). Conclusion Xihuang Pills component-based Chinese medicine can reduce the number of Treg cells by inhibiting the expression of PI3 K/AKT signaling pathway in Treg cells in the tumor microenvironment, and thus exhibit similar anti-cancer effects and mechanisms as Xihuang Pills.
Objective To study whether the Xihuang Pills component-based Chinese medicine has similar anti-cancer effect and mechanism as Xihuang Pills. Methods 4 T1 breast cancer cells were used to establish a tumor model in mice. The model mice were ig administered with Xihuang Pills of low, medium, and high doses (0.39, 0.78 and 1.95 g/kg) and Xihuang Pills component-based Chinese medicine of 0.3 mL (74.5 mg/kg elemene, 2.73 mg/kg taurine, 0.52 mg/kg Muscone and 4.75 mg/kg 11-carbonyl-beta-ethyl ester mastic acid mixture, corresponding to the high dosage group of Xihuang Pills) for 14 d. The tumor tissues were removed,weighed and homogenized to prepare single cell suspensions. Treg cells were prepared by magnetic cell sorting. The changes in number of Treg cells in the tumor microenvironment were detected by flow cytometry and immunohistochemistry. The protein expression of PI3K and AKT in Treg cells in the tumor microenvironment was detected by western blotting. The mRNA expression of PI3 K and AKT in Treg cells in the tumor microenvironment was detected by the Real-time PCR (qRT-PCR). Results Compared with model group and Xihuang Pills low-and medium-dose groups, the weight of tumors was significantly decreased in the Xihuang Pills component-based Chinese medicine group (P < 0.05); The results of flow cytometry and immunohistochemistry showed a significant decrease in the number of Treg cells in the tumor microenvironment (P < 0.05); Western Blotting and qRT-PCR results showed that the expression of PI3 K, AKT protein and their mRNA in Treg cells in the tumor microenvironment was significantly decreased (P < 0.05). Compared with the Xihuang Pills high-dose group, the tumor weight, the number of Treg cells, and the protein expression of PI3 K and AKT and their mRNA expression in the Xihuang Pills component-based Chinese medicine group were significant increased (P < 0.05). Conclusion Xihuang Pills component-based Chinese medicine can reduce the number of Treg cells by inhibiting the expression of PI3 K/AKT signaling pathway in Treg cells in the tumor microenvironment, and thus exhibit similar anti-cancer effects and mechanisms as Xihuang Pills.
引文
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[18]苗晋鑫,苗明三.组分中药研究现状、存在问题及思考[J].中医学报,2012,27(9):1116-1119.
[2]王硕,刘杰,林洪生.中医药在癌症治疗中的作用[J].环球中医药,2013,6(1):31-35.
[3]张俊华,樊官伟,张晗,等.组分中药理论的发展与应用[J].中国中药杂志,2017,42(21):4054-4058.
[4]岳广欣,刘丽梅,柏冬,等."组分中药"药理研究内容和特点述评[J].中国中医药信息杂志,2015,22(3):121-126.
[5]谭萍.西黄丸治疗恶性肿瘤应用体会[J].浙江中西医结合杂志,2001,11(4):244-244.
[6]张霞丽.牛磺酸对肿瘤细胞凋亡的作用及其机制初步研究[D].南昌:南昌大学,2012.
[7]刘永惠,常靖,薛陆军,等.麝香酮对血瘀证乳腺癌生长及对VEGF表达的影响[J].西安交通大学学报:医学版,2014,35(4):547-550.
[8]Yadav V R,Prasad S,Sung B,et al.Boswellic acid inhibits growth and metastasis of human colorectal cancer in orthotopic mouse model by downregulating inflammatory,proliferative,invasive and angiogenic biomarkers[J].Int J Cancer,2012,130(9):2176-2184.
[9]麻杰,陈娟,赵冰洁,等.抗癌药物β-榄香烯及其衍生物的研究进展[J].中草药,2018(5):1184-1191.
[10]Li X Y,Su L,Jiang Y M,et al.The antitumor effect of xihuang pill on treg cells decreased in tumor microenvironment of 4T1 breast tumor-bearing mice by PI3K/AKT~AP-1 signaling pathway[J].Evid Based Compl Alter Med,2018,2018:6714829.
[11]谷伟伟,叶韵斌.调节性T细胞与肿瘤相互作用机制的研究进展[J].医学综述,2012,18(5):680-684
[12]Liu F F,Lang R G,Zhao J,et al.CD8?cytotoxic T cell and FOXP3?regulatory T cell infiltration in relation to breast cancer survival and molecular subtypes[J].Breast Cancer Res Treat,2011,130(2):645-655.
[13]戴晓晓,张永胜,刘加豪,等.乳腺癌流行病学及临床病理学分析[J].中国医院药学杂志,2018:1-5.
[14]吴世婷,刘丽芳,丁玲,等.中医药治疗乳腺癌临床研究进展[J].湖南中医杂志,2015,31(9):170-173.
[15]Sami A,Karsy M.Targeting the PI3K/AKT/mTORsignaling pathway in glioblastoma:novel therapeutic agents and advances in understanding[J].Tumour Biol,2013,34(4):1991-2002.
[16]李建,周怀君.PI3K/Akt信号通路在肿瘤研究中的进展[J].东南大学学报:医学版,2014,33(3):392-395.
[17]郭倩,田成旺,任涛,等.中药药效物质基础研究进展[J].世界科学技术:中医药现代化,2015,17(3):648-654.
[18]苗晋鑫,苗明三.组分中药研究现状、存在问题及思考[J].中医学报,2012,27(9):1116-1119.