臭牡丹提取物对H22荷瘤小鼠体内抑瘤作用及对相关蛋白表达的影响
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摘要
目的研究臭牡丹提取物对H22实体移植瘤的抑制作用及与B淋巴细胞瘤/白血病-2(Bcl-2)和Bcl-2相关X蛋白(Bax)表达的相关性。方法皮下接种H22瘤株建立小鼠H22实体移植瘤模型。60只荷瘤鼠随机分为模型组、5-氟尿嘧啶(5-Fu)组和臭牡丹高、中、低剂量(2.18、1.09、0.55 g/kg)组,每组12只,各组分别给予生理盐水、5-Fu(25 mg/kg,1次/d)及不同浓度的臭牡丹提取物干预。连续给药14 d后处死,检测小鼠体质量变化、抑瘤率及脾脏和胸腺指数;Western blot法检测Bcl-2和Bax蛋白的表达水平。结果与模型组比较,臭牡丹高、中剂量组小鼠的肿瘤质量均显著降低(P<0.01),胸腺和脾脏指数显著增加(P<0.05);同时Bax蛋白的表达水平显著上调,Bcl-2蛋白的表达显著降低(P<0.01)。结论臭牡丹提取物能够抑制H22实体移植瘤生长,其机制可能与上调Bax/Bcl-2比值和免疫调节有关。
Objective To study the inhibitory effect of Clerodendron bungei extract on H22 solid xenograft and its correlation with the expression of B cell lymphoma/leukemia-2(Bcl-2) and Bcl-2 related X protein(Bax) proteins.Methods A mouse H22 solid xenograft model was established by subcutaneous inoculation of H22 tumor strain.Sixty tumor-bearing mice were randomly divided into model group,5-fluorouracil(5-Fu) group and Clerodendron bungei high,medium and low dose groups,12 rats in each group.Each group was given normal saline and 5-Fu(25 mg/kg,1 time/d) and different concentrations of Clerodendron bungei intervention.After 14 days of continuous administration,the tumor was sacrificed,and the changes in body mass,tumor inhibition rate and spleen and thymus index were measured.Western blot was used to detect the expression levels of Bcl-2 and Bax proteins.Results Compared with the model group,the tumor mass of the high,middle and low dose groups of the Clerodendron Bungei group was significantly decreased(P<0.01),and the thymus and spleen index increased(P<0.05).At the same time,the expression level of bax protein was up-regulated and the expression of bcl-2 protein was decreased(P<0.01).Conclusion The Clerodendron bungei extract can inhibit the growth of H22 solid xenografts,and its mechanism may be related to up-regulating the ratio of bax/bcl-2 to induce apoptosis.
Objective To study the inhibitory effect of Clerodendron bungei extract on H22 solid xenograft and its correlation with the expression of B cell lymphoma/leukemia-2(Bcl-2) and Bcl-2 related X protein(Bax) proteins.Methods A mouse H22 solid xenograft model was established by subcutaneous inoculation of H22 tumor strain.Sixty tumor-bearing mice were randomly divided into model group,5-fluorouracil(5-Fu) group and Clerodendron bungei high,medium and low dose groups,12 rats in each group.Each group was given normal saline and 5-Fu(25 mg/kg,1 time/d) and different concentrations of Clerodendron bungei intervention.After 14 days of continuous administration,the tumor was sacrificed,and the changes in body mass,tumor inhibition rate and spleen and thymus index were measured.Western blot was used to detect the expression levels of Bcl-2 and Bax proteins.Results Compared with the model group,the tumor mass of the high,middle and low dose groups of the Clerodendron Bungei group was significantly decreased(P<0.01),and the thymus and spleen index increased(P<0.05).At the same time,the expression level of bax protein was up-regulated and the expression of bcl-2 protein was decreased(P<0.01).Conclusion The Clerodendron bungei extract can inhibit the growth of H22 solid xenografts,and its mechanism may be related to up-regulating the ratio of bax/bcl-2 to induce apoptosis.
引文
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[29]陈璇,童晔玲,任泽明,等.杨梅醇对人肝癌HepG2细胞Caspase-9、Bcl-2、p21和Bax表达水平的影响[J].中华中医药杂志,2015,30(10):3656-3658.
[2]朱克俭,张亚利,欧阳剑虹,等.保肺饮预防肺癌的临床流行病学研究[J].中国中医药科技,1997(6):332-333.
[3]陈思勤,朱克俭,程晓燕,等.臭牡丹化学成分及其药理作用研究进展[J].湖南中医杂志,2012,28(2):141-142.
[4]洪梓德,莫志贤.中药抗肿瘤机制中的11种信号通路[J].中国实验方剂学杂志,2018,24(21):205-218.
[5]蔡利,管翰粟.中药在抗肿瘤过程中的应用及研究进展[J].中医临床研究,2017,9(11):141-142.
[6]白云,刘萍,贾博宇,等.中药抗肿瘤作用的研究进展[J].中国药师,2014,17(8):1406-1409.
[7]余娜,朱克俭,马思静,等.臭牡丹总黄酮对A549细胞增殖迁移侵袭力及Wnt通路相关蛋白的影响[J].世界中医药,2018,13(4):978-983,987.
[8]余娜,朱克俭,马思静,等.臭牡丹总黄酮通过调控Wnt/β-catenin通路影响A549细胞上皮间质转化研究[J].中草药,2018,49(3):663-670.
[9]陈思勤,朱克俭,李勇敏,等.臭牡丹总黄酮抑制小鼠Lewis肺癌实体瘤及其与p53、bcl-2、bax表达相关性研究[J].世界中医药,2016,11(6):946-949.
[10]胡琦,朱克俭,谭小宁,等.臭牡丹总黄酮对人肝癌HepG2细胞增殖作用的实验研究[J].湖南中医杂志,2015,31(4):166-168.
[11]胡琦,谭小宁,余娜,等.臭牡丹总黄酮介导Wnt/β-catenin信号转导诱导人肝癌细胞HepG2的凋亡[J].世界中医药,2016,6(2):954-957.
[12]Babichev Y,Kabaroff L,Datti A,et al.PI3K/AKT/mTORinhibition in combination with doxorubicin is an effective therapy for leiomyosarcoma[J].J Transl Med,2016,14(1):67-71.
[13]Cho D C.Targeting the PI3K/Akt/mTOR pathway in malignancy:rationale and clinical outlook[J].Bio Drugs,2014,28(4):373-381.
[14]马星,夏伟.中药抗肝癌细胞增殖和诱导细胞凋亡的信号通路研究进展[J].上海中医药杂志,2018,52(8):98-101.
[15]丁大伟,章永红.以细胞凋亡通路为靶点的抗肿瘤中药研究进展[J].中国老年学杂志,2018,38(1):239-241.
[16]周红林,刘建新,周俐,等.臭牡丹提取物抗炎镇痛抗过敏作用的实验研究[J].中国新药杂志,2006,15(23):2027-2029.
[17]李慧乐,莫传伟,赵春辉,等.莪术提取物榄香烯对肝癌H22荷瘤小鼠的抑瘤作用[J].中国临床药理学杂志,2018,34(11):1345-1348.
[18]Su T S,Lu H Z,Cheng T,et al.Long-term survival analysis in combined transarterial embolization and stereotactic body radiation therapy versus stereotactic body radiation monotherapy for unresectable hepatocellular carcinoma>5 cm[J].BMC Cancer,2016,16(1):834.
[19]诸佳瑜,陈闯,欧杰,等.中医辩证治疗原发性肝癌的研究进展[J].当代医药论丛,2017,15(10):32-34.
[20]Liang S H,Sun K,Wang Y,et al.Role of Cyt-C/caspases-9,3,Bax/Bcl-2 and the FAS death receptor pathway in apoptosis induced by zinc oxide nanoparticles in human aortic endothelial cells and the protective effect by alphalipoic acid[J].Chem Biol Interact,2016,258(1):40-51.
[21]Dou C Q,Han M M,Zhang B,et al.Chrysotoxene induces apoptosis of human hepatoblastoma HepG2 cells in vitro and in vivo via activation of the mitochondriamediated apoptotic signaling pathway[J].Oncol Lett,2018,15(4):4611-4618.
[22]Lin H F,Hsieh M J,Hsi Y T,et al.Celastrol-induced apoptosis in human nasopharyngeal carcinoma is associated with the activation of the death receptor and the mitochondrial pathway[J].Oncol Lett,2017,14(2):1683-1690.
[23]徐云峰,刘松坚.Bcl-2家族的研究进展[J].中国疗养医学,2014,23(10):884-886.
[24]Li H L,Lv B,Kong L,et al.Nova1 mediates resistance of rat pheochromocytoma cells to hypoxia-induced apoptosis via the Bax/Bcl-2/caspase-3 pathway[J].Int JMol Med,2017,40(4):1125-1133.
[25]李扩,许秋然,刘欣,等.miR-204通过下调Bcl-2和Sirt1表达抑制肝癌细胞生长[J].细胞与分子免疫学杂志,2015,31(2):168-172.
[26]Williams M M,Cook R S.Bcl-2 family proteins in breast development and cancer:could Mcl-1 targeting overcome therapeutic resistance[J].Oncotarget,2015,6(6):3519-3130.
[27]李文婷,赵凤鸣,戴紫函,等.白花蛇舌草注射剂抑制人肝癌细胞SMMC-7721增殖及调控Bcl-2/CytC信号通路诱导线粒体凋亡[J].中成药,2015,37(10):2264-2267.
[28]崔明宇,崔轶琼,刘媛媛,等.香叶木苷对人肝癌HepG2细胞凋亡及Bcl-2/Bax基因表达的影响[J].药物评价研究,2018,41(11):1958-1962.
[29]陈璇,童晔玲,任泽明,等.杨梅醇对人肝癌HepG2细胞Caspase-9、Bcl-2、p21和Bax表达水平的影响[J].中华中医药杂志,2015,30(10):3656-3658.