穿心莲内酯对小鼠乳腺癌的抑制作用和机制  

The anti-tumor activity of andrographolide on breast cancer of mice and its mechanism

作　　者：鹿蓓蓓 傅鹤秀 Eric Achiborebador Okrah 郭文洁[2] 高静[1] LU Bei-bei;FU He-xiu;ERIC Achiborebador Okrah;GUO Wen-jie;GAO Jing(Lab of Mitochondria and Neurobiology,School of Pharmacy,Jiangsu University,Zhenjiang Jiangsu 212013,China;School of Life Sciences,Nanjing University,Nanjing 210023,China)

机构地区：[1]江苏大学药学院,线粒体与神经生物学实验室,江苏镇江212013 [2]南京大学生命科学学院,江苏南京210023

出　　处：《中国药理学通报》2022年第1期26-32,共7页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81402938)。

摘　　要：目的研究穿心莲内酯(andrographolide, Andro)对乳腺癌小鼠肿瘤生长的抑制作用及对线粒体功能的影响并探讨其机制。方法 MTT法检测Andro对4T1的抑制作用;测量肿瘤体积,计算脏器指数,HE染色分析Andro对肿瘤的抑制作用及对免疫、代谢器官的影响。试剂盒检测血清中乳酸含量,分析Andro对糖酵解的影响;试剂盒检测肿瘤组织中ATP、乙酰辅酶A含量、Western blot检测PDH蛋白表达,分析Andro对氧化磷酸化的影响;Clark氧电极、JC-1染色检测耗氧量及线粒体膜电位,分析线粒体功能。结果 Andro可抑制4T1细胞增殖,抑制肿瘤生长,且对免疫、代谢器官无显著影响;Andro可下调乳酸含量,抑制糖酵解;上调ATP、乙酰辅酶A含量及PDH蛋白表达,恢复氧化磷酸化;升高耗氧及线粒体膜电位,改善线粒体功能。结论 Andro对乳腺癌小鼠肿瘤生长具有抑制作用,其机制可能与抑制糖酵解,恢复氧化磷酸化,改善线粒体功能有关。Aim To evaluate the effects of andrographolide(Andro) on suppressing tumor growth and improving mitochondrial function on mouse breast cancer and explore its mechanism. Methods MTT assay was performed to measure the effect of Andro on the growth capacity of mouse breast cancer cell line 4 T1. Mice were treated with Andro, then tumor volume measured, organ index calculated and Hematoxylin-eosin(HE)stained to detect the inhibitory effect of Andro on tumors. Lactate assay kit was used to detect the lactate level in mouse serum to check glycolysis discrepancy.To illustrate the transformation in oxidative phosphorylation(OXPHOS), Acetyl coenzyme A(Acetyl-CoA)assay kit was used to ascertain Acetyl-CoA content level, Western blot was used to detect the protein expression of pyruvate dehydrogenase(PDH), and ATP assay kit was used to ascertain ATP content level. The mitochondria functions were analyzed: oxygen( O;)consumption was measured by Clark oxygen electrode,and mitochondrial membrane potential was detected by JC-1 staining.Results Andro could effectively inhibit the proliferation of 4 T1 cells and the growth of tumors,and had no significant damage on normal organs. Andro reduced serum lactic acid content, indicateing that Andro inhibited the process of glycolysis. The expression of PDH, content of acetyl-CoA and ATP content increased with the increase of Andro concentration. It showed that Andro could up-regulate oxidative phosphorylation. In addition, the O;consumption and mitochondrial membrane potential increased in 4 T1 cells,indicating that Andro could recover mitochondrial function.Conclusions Andro can inhibit the growth of mouse breast cancer, and its mechanism may be related to the inhibition of glycolysis level, restoration of OXPHOS and improvement of mitochondrial function.

关 键 词：穿心莲内酯 乳腺癌 4T1荷瘤小鼠 糖酵解 氧化磷酸化 线粒体 

分 类 号：R-332[医药卫生] R284.1