淫羊藿苷对5-氟尿嘧啶诱导的大鼠骨髓间充质干细胞损伤的保护作用  

Protection of icariin on impairment of rat bone mesenchymal stem cells induced by 5-fluorouracil in vitro

作　　者：张小年[1] 张洁文[2] 吴绍锋 雷清贵 郭宇琳 刁远明[1] ZHANG Xiao-nian;ZHANG Jie-wen;WU Shao-feng;LEI Qing-gui;GUO Yu-lin;DIAO Yuan-ming(College of Basic Medical Science, Guangzhou University of Chinese Medicine, Guangzhou 510006, China;Dept of Traditional Chinese Medicine, the First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080, China;College of Traditional Chinese Medicine, Guangzhou University of Chinese Medicine, Guangzhou 510006, China)

机构地区：[1]广州中医药大学基础医学院,广东广州510006 [2]中山大学附属第一医院中医科,广东广州510080 [3]广州中医药大学中药学院,广东广州510006

出　　处：《中国药理学通报》2020年第5期675-681,共7页Chinese Pharmacological Bulletin

基　　金：广东省高等学校优秀青年教师培养计划(No YQ2015042);广东省科技计划(No 2015A070706007);广东省中医药局中医药科研项目(No 20191095)。

摘　　要：目的探讨淫羊藿苷(icariin,ICA)对5-氟尿嘧啶(5-Fluorouracil,5-FU)所致大鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)损伤的保护作用及其机制。方法 MTT比色法检测细胞存活率,Hoechst 33258染色法观察细胞凋亡形态,Annexin V-FITC/PI双染流式检测细胞早期凋亡率,Western blot法检测Cleaved caspase-3蛋白表达水平,caspase-3活性检测试剂盒检测caspase-3酶活性,一氧化氮合酶(nitric oxide synthase,NOS)测定试剂盒检测iNOS、TNOS含量。结果 MTT检测发现一定浓度的ICA可促进BMSCs增殖,并可对抗5-FU对大鼠BMSCs造成的生长抑制,明显提高细胞存活率; 10,20μmol·L-1ICA与2. 5×10-2g·L-15-FU共同作用大鼠BMSCs 48 h后,与模型组比较,大鼠BMSCs的凋亡率明显降低(P <0. 01),Cleaved caspase-3蛋白表达量以及caspase-3酶活性下降(P <0. 01),iNOS和TNOS活力有一定程度的降低(P <0. 05)。结论ICA可以抑制5-FU对BMSCs所造成的损伤作用,保护BMSCs细胞,其作用机制可能是降低iN OS和TNOS的活性,抑制caspase-3激活,从而抑制BMSCs凋亡的发生。Aim To study the protective effect of icariin on the impairment of rat bone mesenchymal stem cells(BMSCs)induced by 5-fluorouracil(5-FU)and the underlying mechanism.Methods The proliferation of BMSCs was evaluated by MTT assay,the morphological change of apoptotic cells was tested by using the chromatin dye Hoechst 33258,and the apoptotic rate was analyzed by flow cytometry.The proteins expression of cleaved caspase-3 was detected with Western blotting,the activity of caspase-3 was detected by Activity Assay Kit;iNOS and TNOS were detected by nitric oxide synthase(NOS)assay kit.Results MTT detection showed that 5-FU could reduce the survival rate of BMSCs,10,20,40μmol·L^-1 ICA could promote the proliferation of BMSCs,and blocked the inhibition of BMSCs cells growth induced by 5-FU.When BMSCs cells were treated by both ICA(10,20μmol·L^-1)and 5-FU(2.5×10-2 g·L^-1)for 48 hours,compared with model group,the early apoptotic rates of ICA test groups were significantly reduced(P<0.01),the expression of cleaved caspase-3 and the activity of caspase-3 were significantly reduced(P<0.01),and the activity of iNOS and TNOS was inhibited in a certain degree(P<0.05).Conclusions ICA could protect BMSCs against 5-FU-induced damage.The protective role of ICA is probably related to reducing the activity of iNOS and TNOS,inhibiting the activation of caspase-3,and inhibiting the occurrence of apoptosis.

关 键 词：淫羊藿苷 5-氟尿嘧啶 骨髓间充质干细胞 化疗损伤 凋亡 保护 

分 类 号：R-332[医药卫生] R284.1