红黄煎剂通过调节AKT/FoxO3a通路改善阿霉素造成的心肌细胞凋亡  

Honghuang Decoction Enhances Protective Effect Against Doxorubicin-Induced Myocardial Apoptosis Via Akt/FoxO3a Pathway

作　　者：薛静娴[1] 朱智媛 章宜芬[1] 卞卫和[1] 姚昶[1] XUE Jingxian;ZHU Zhiyuan;ZHANG Yifen;BIAN Weihe;YAO Chang(The Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210046,Jiangsu,China)

机构地区：[1]南京中医药大学附属医院

出　　处：《中华中医药学刊》2018年第10期2397-2401,I0017,共6页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(81473676);江苏省高校优势学科建设项目II期项目(012062003010);江苏省中医院高峰学术人才培养项目(y2014cr06)

摘　　要：目的：阿霉素是一种临床常用的化疗药物,其心脏毒性制约了它的临床使用,为了了解红黄煎剂在体外对阿霉素造成的心脏损伤降的作用,并探讨机制,做如下体外实验。方法：体外培养H9c2心肌细胞。用不同浓度红黄煎剂（HHD）,阿霉素（DOX）,红黄煎剂＋阿霉素分别处理H9c2心肌细胞24 h,MTT法检测细胞活力,TUNEL和DAPI双染法检测细胞凋亡率,Western Blot法检测细胞凋亡相关蛋白。然后Si-Fox O3a转染H9c2心肌细胞,免疫荧光及Western Blot法确定转染效果,western及MTT法检测红黄煎剂,阿霉素,红黄煎剂＋阿霉素对Si-Fox O3a转染的H9c2心肌细胞的作用。结果：不同浓度（0-2 mg·m L-1）的红黄煎剂作用与H9c2细胞,发现对细胞有一定的促进增值的作用（P 〈0. 05）,阿霉素作用与H9c2细胞,其IC 50为（3. 03±0. 11）μM,加用红黄煎剂以后发现其抑制曲线下移,其IC 50为（7. 07±0. 48）μM,其差异有统计学意义（P 〈0. 01）。TUNEL＆DAPI双染法检测细胞凋亡,发现HHD＋DOX将单用DOX组的凋亡率从（43. 23±5. 22）%下调到（25. 22±3. 21）%。其差异有统计学意义（P 〈0. 01）。红黄煎剂能显著下调阿霉素引起的Caspase-3的激活（P 〈0. 05）,同时能上调生存蛋白Bcl-2（P 〈0. 05）。同时红黄煎剂能够提高H9c2细胞中的Akt/Fox O3a磷酸化的水平。转染siFox O3a后细胞增值率下降（P 〈0. 05）,并且减弱了HHD促进细胞增值的作用（P 〈0. 05） Western Blot检测也证明了HHD在转染细胞系中的保护作用降低（P 〈0. 05）。结论：红黄煎剂能降低阿霉素诱导的心肌细胞凋亡,其可能是通过调节AKT/Fox O3a产生的。Objectie：Doxorubicin（ DOX）,an anthracycline-based antitumor drug,has been used in clinic. However,its cardiac toxicity limits its clinical use. Honghuang Decoction（ HHD） is a formula that has been used to treat doxorubicin-induced cardiotoxicity for many years. Therefore,in this study,we aimed to investigate the underlying mechanisms in vitro. Methods： H9 c2 cells were randomly assigned to three groups and treated with doxorubicin,Honghuang Decoction,respectively. Cell growth was determined by the MTT cell proliferation and cytotoxicity assay kit. Cell apoptosis was assessed by using TUNEL and DAPI staining. The expression of c-caspase 3/caspase 3,bcl-2,Fox O3 a,and AKT was examined by Western blotting. The Fox O3 a gene in H9 c2 cells was knocked down by small interfering RNA（siRNA） to prove that HHD reduced cell apoptosis induced by doxorubicin via AKT/Fox O3 a pathway. Results： Our study proved the HHD reduced cell apoptosis in H9 c2 cell lines. This protective effect was achieved by HHD via Akt/Fox O3 a pathway. Conclusion： It can be suggested that HHD had the protective effect of doxorubicin-induced myocardial apoptosis via Akt/Fox O3 a pathway.

关 键 词：红黄煎剂 阿霉素 H9C2细胞 AKT FOXO3A 心肌细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]