黄芪建中汤含药血清联合顺铂对A549细胞的抑制作用及Smad3、Smad7表达的影响
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摘要
目的:观察黄芪建中汤含药血清联合顺铂对肺癌A549细胞中Smad3、Smad7蛋白表达的影响。方法:应用MTT法分别计算5%高、中、低剂量和10%高、中、低剂量黄芪建中汤含药血清作用于人肺腺癌A549细胞株24、48、72 h的半数抑制浓度(IC_(50))、黄芪建中汤最佳含药血清(10%中剂量,48h)联合最佳顺铂(5μmol/mL,48 h)作用于人肺腺癌A549细胞株的增殖抑制率。免疫组织化学显色技术检测肺癌A549细胞内Smad3、Smad7蛋白的表达情况;Western Blot技术检测Smad3、Smad7蛋白表达。结果:①黄芪建中汤含药血清作用于A549细胞株48h的(IC_(50))为10%中剂量。黄芪建中汤含药血清联合顺铂作用于A549细胞的增殖抑制率高于两单药组和空白组(P<0.05)。②免疫组化方法检测Smad3、Smad7在A549细胞中的表达,与其它组相比在中西药联合组(HuangQi+Cisplatin)中Smad3蛋白表达棕黄色颗粒最少,Smad7蛋白表达棕黄色颗粒最多。差异具有统计学意义(P<0.05)。③Western Blot技术检测Smad3与Smad7的表达。与其它组相比,中西药联合组Smad3表达下调,Smad7表达上调(P<0.05)。结论:黄芪建中汤含药血清联合顺铂对肺癌A549细胞具有抑制增殖作用,其作用机制可能与上调smad7蛋白表达,下调Smad3蛋白表达有关。
Objecttive:To observe the effect of HuangqiJianzhong Decoction combined with cisplatin on the expressions of Smad3 and Smad7 proteins in lung cancer A549 cells. Methods:By MTT method,we calculated half inhibitory concentration(IC_(50)) of the drug serum of 5% and 10% HuangqiJianzhong Decoctionon human lung adenocarcinoma A549 cell line for 24, 48, 72 h and the proliferation inhibition rate of serum-containing HuangqiJianzhong Decoction(10%,middle dose, 48 h) and cisplatin(5 μmol/mL, 48 h) on human lung adenocarcinoma A549 cell line. The expression and localization of Smad3 and Smad7 proteins in lung cancer A549 cells were detected by immunohistochemical coloration. Protein expressions of Smad3 and Smad7 were detected by Western Blot. Results: The serum of Huangqi Jianzhong Decoctionon A549 cell linefor 48 h was 10% medium dose. The proliferation inhibition rate of Huangqi Jianzhong Decoction combined with cisplatin on A549 cells was higher than that of the two monotherapy groups and the blank group(P<0.05). The expressions of Smad3 and Smad7 in A549 cells were detected by immunohistochemistry. Compared with other groups, the brown-yellow particles were leastly expressed by Smad3 protein in the combination of Chinese and western medicine(Huangqi+Cisplatin), and the brown-yellow particles were mostly expressed by Smad7 protein. The difference was statistically significant(P<0.05). The expressions of Smad3 and Smad7 were detected by Western Blot. Compared with other groups, Smad3 expression was down-regulated and Smad7 expression was up-regulated(P<0.05). Conclusion: Huangqi Jianzhong Decoction combined with cisplatin can inhibit the proliferation of lung cancer A549 cells, and the mechanism may be related to up-regulation of smad7 protein expression and down-regulation of Smad3 protein expression.
Objecttive:To observe the effect of HuangqiJianzhong Decoction combined with cisplatin on the expressions of Smad3 and Smad7 proteins in lung cancer A549 cells. Methods:By MTT method,we calculated half inhibitory concentration(IC_(50)) of the drug serum of 5% and 10% HuangqiJianzhong Decoctionon human lung adenocarcinoma A549 cell line for 24, 48, 72 h and the proliferation inhibition rate of serum-containing HuangqiJianzhong Decoction(10%,middle dose, 48 h) and cisplatin(5 μmol/mL, 48 h) on human lung adenocarcinoma A549 cell line. The expression and localization of Smad3 and Smad7 proteins in lung cancer A549 cells were detected by immunohistochemical coloration. Protein expressions of Smad3 and Smad7 were detected by Western Blot. Results: The serum of Huangqi Jianzhong Decoctionon A549 cell linefor 48 h was 10% medium dose. The proliferation inhibition rate of Huangqi Jianzhong Decoction combined with cisplatin on A549 cells was higher than that of the two monotherapy groups and the blank group(P<0.05). The expressions of Smad3 and Smad7 in A549 cells were detected by immunohistochemistry. Compared with other groups, the brown-yellow particles were leastly expressed by Smad3 protein in the combination of Chinese and western medicine(Huangqi+Cisplatin), and the brown-yellow particles were mostly expressed by Smad7 protein. The difference was statistically significant(P<0.05). The expressions of Smad3 and Smad7 were detected by Western Blot. Compared with other groups, Smad3 expression was down-regulated and Smad7 expression was up-regulated(P<0.05). Conclusion: Huangqi Jianzhong Decoction combined with cisplatin can inhibit the proliferation of lung cancer A549 cells, and the mechanism may be related to up-regulation of smad7 protein expression and down-regulation of Smad3 protein expression.
引文
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[21] 王莉,余刚,向德兵,等.肺鳞癌组织中TβRⅡ、Smad7的表达及其相关性[J].肿瘤学杂志,2010,16(12):926-929.
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[2] 杨冲,李宪松,刘孟军,等.酸枣的营养成分及开发利用研究进展 [J].北方园艺,2017,41(5):184-188.
[3] 周金玲,杨玉芳,黄振光,等.三七总皂苷通过 HIF-1α/BNIP3 途径增强线粒体自噬保护大鼠顺铂肾损伤 [J].中国药学杂志,2017,52(3):196-200.
[4] 丁斗,董小君,王敏,等.尿β2-微球蛋白在顺铂所致肾毒性中的早期监控 [J].甘肃中医学院学报,2013,30(2):5-7.
[5] 魏自太,王彩云,包素珍,等.脾气虚证肺癌小鼠血清IL-12,IL-10的表达及加味黄芪建中汤的影响[J].云南中医学院学报,2017,40(4):10-13.
[6] 包素珍,张誉引,陈淇.加味黄芪建中汤对脾虚证肺癌小鼠VEGF的表达及MVD的影响[J].陕西中医药大学学报,2016,39(1):99-101.
[7] 侯星宇.基于隐喻认知的癌症放化疗术后中医防护思想与方法研究[D].北京:北京中医药大学,2018.
[8] 周明德.基于药物干预的慢性乙型肝炎肝郁脾虚证特征性蛋白的验证研究[D].成都:成都中医药大学,2017.
[9] 张莹.黄芪建中汤加味对中晚期非小细胞肺癌患者化疗期间免疫功能和生存质量的影响[J].河南中医,2015,35(3):474-476.
[10] 罗蒙蒙.黄芪建中汤加减防治晚期非小细胞肺癌化疗毒副反应的临床研究[D].郑州:河南中医药大学,2017.
[11] TGF-β超家族调节着各种细胞的生长和分化,对于抑制细胞增殖、肿瘤发生及发展的过程起到重要的作用.
[12] 吕洋.TGF-β1诱导、转染大鼠骨髓间充质干细胞启动心肌分化及抑制Wnt/β-catenin信号通路的调控机制[D].石家庄:河北医科大学,2018.
[13] 张乐.秦川牛Smad2和Smad3基因转录调控研究[D].咸阳:西北农林科技大学,2018.
[14] Verechia F,Mauviel A.Transforming growth factor-βsignaling through the Smad pathway:role in extracelular matrix gene expresion and regula- tion [J].J Invest Dermatol,2002,118(2):211- 215.
[15] 付玲珠,郑婷,张永生.TGF-β/Smad信号转导通路与肝纤维化研究进展[J].中国临床药理学与治疗学,2014,19(10):1189-1195.
[16] 刘镕,赵琴平,董惠芬,等.TGF-β信号传导通路及其生物学功能[J].中国病原生物学杂志,2014,9(1):77-83.
[17] 苗小玲.四逆散加味对肝纤维化大鼠TGF-β/Smads通路及P-STAT3、P38MAPK、a-SMA的影响[D].郑州:河南中医学院,2013.
[18] 高杨,阎继东,张志勇,等.BCAR1和Smad3蛋白在非小细胞肺癌组织中的表达及临床病理意义[J].中国老年学杂志,2013,33(4):800-802.
[19] 吴芙蓉,姜玲,何晓丽,等.橙皮苷对肝星状细胞TGF-β1/Smad信号通路的影响[J].中国中药杂志,2015,40(13):2639-2643.
[20] 陈炎.TGF-β信号中R-Smads磷酸酶在宫颈癌发生发展过程中的表达及作用机制研究[D].合肥:安徽医科大学,2018.
[21] 王莉,余刚,向德兵,等.肺鳞癌组织中TβRⅡ、Smad7的表达及其相关性[J].肿瘤学杂志,2010,16(12):926-929.
[22] 林艳蓝.基于TGF-β1/Smad3信号通路研究桃核承气汤对CRF大鼠肾间质纤维化中miR-29表达的影响[D].福州:福建中医药大学,2018.
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