摘要
目的:以有丝分裂原激活蛋白激酶(p38)/信号传导与转录激活因子3(STAT3)信号通路为基础,探究阳和汤含药血清对乳腺癌MCF-7细胞凋亡的影响。方法:制备含生药1 g·m L^-1的阳和汤药液,将SD大鼠随机分为空白组(蒸馏水),阳和汤高、中、低剂量组(24,12,6 g·kg^-1),灌胃后制备阳和汤含药血清,用10%含药血清干预MCF-7细胞,通过细胞增殖及细胞毒性实验(CCK-8)检测阳和汤含药血清对MCF-7细胞增殖的影响;通过流式细胞术检测MCF-7细胞的凋亡情况;通过蛋白免疫印迹法(Western blot)检测p38和STAT3蛋白的表达情况;通过实时荧光定量聚合酶链式反应(Real-time PCR)检测B细胞淋巴瘤/白血病-xl(Bcl-xl)及生存素(Survivin)mRNA表达的情况。结果:CCK-8实验显示,与空白组比较,阳和汤含药血清抑制MCF-7细胞增殖,且呈时间与剂量依赖性。其中高剂量组抑制作用最为明显,不同时间抑制率分别达到38%,45%,54%(P<0.01);流式细胞术实验表明,与空白组比较,阳和汤含药血清中、高剂量组细胞凋亡率明显增加,且呈剂量依赖性,凋亡率分别达11.6%,16.5%(P<0.05,P<0.01);Western blot表明,与空白组比较,阳和汤中、高剂量含药血清组p38,STAT3蛋白表达减少(P<0.01),且呈剂量依赖性;Real-time PCR实验表明,与空白组比较,阳和汤中、高剂量含药血清组Bcl-xl,Survivin mRNA表达减少(P<0.05,P<0.01),且呈剂量依赖性。结论:阳和汤含药血清可以促进乳腺癌MCF-7细胞的凋亡,可能与p38/STAT3信号通路有关。
Objective:To investigate the effect of serum containing Yanghetang(YHT)on the apoptosis of MCF-7 cells in breast cancer based on the mitogen-activated protein kinase(p38)/signal transduction and transcriptional activator 3(STAT3)signal pathway.Method:YHT liquid with crude drug 1 g·m L^-1was prepared.Female SD rats were randomly divided into control group(distilled water),and high,medium and lowdose YHT groups(24,12,6 g·kg^-1).YHT-medicated serum was prepared,and 10%medicated serum was used to intervene MCF-7 cells.Cell proliferation and cytotoxicity assay(CCK-8)was used to detect the effect of serum containing YHT on MCF-7 cell proliferation,apoptosis of MCF-7 cells was detected by flow cytometry protein expressions of p38 and STAT3 were detected by Western blot,Quantitative Real-time PCR(Real-time PCR)was used to detect the expressions of B-cell lymphoma/leukemia-xl(Bcl-xl)and Survivin mRNA.Result:CCK-8assay showed that YHT serum inhibited the proliferation of MCF-7 cells in a time and dose-dependent manner compared with the blank group.The inhibitory effect was most obvious in the high-dose group,with the inhibition rates of 38%,45%and 54%at different time points(P<0.01).Flow cytometry showed that,compared with the blank group,the apoptosis rate in the medium and high-dose groups increased significantly in a dose-dependent manner,with the apoptosis rates at 11.6%and 16.5%respectively(P<0.05,P<0.01).Western blot analysis showed that,compared with the blank group,the expressions of p38 and STAT3 protein was decreased in high,medium-dose YHT groups(P<0.01)in a dose-dependent manner.Compared with the blank group,the expressions of Bcl-xl and Survivin mRNA were decreased in high,medium-dose YHT groups(P<0.05,P<0.01)in a dose-dependent manner.Conclusion:YHT serum can promote the apoptosis of MCF-7 cells in breast cancer,which may be related to the p38/STAT3 signaling pathway.
作者
杨硕
李康乐
朱中博
黄芊
姚佳
职媛
窦建卫
YANG Shuo;LI Kang-le;ZHU Zhong-bo;HUANG Qian;YAO Jia;ZHI Yuan;DOU Jian-wei(School of Basic Medicine,Gansu University of Chinese Medicine,Lanzhou 730000,China;School of Pharmacy,Xi'an Jiaotong University,Xi'an 710061,China;Xi'an Hospital of Traditional Chinese Medicine,Xi'an 710021,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2020年第5期6-10,共5页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(81873311,81573983)
西安市卫生健康委员会课题项目(SZJ201907).
