新橙皮苷对化疗损伤骨髓间充质干细胞的保护作用研究  

Protective Effects of Neohesperdin on Mouse Bone Marrow Mesenchymal Stem Cells against Chemotherapy Damage Induced by 5-Fluorouracil

作　　者：刁远明[1] 张小年[1] 周贤熙 余丽娟[1] 郭宇琳 雷清贵 DIAO Yuanming;ZHANG Xiaonian;ZHOU Xianxi;YU Lijuan;GUO Yulin;LEI Qinggui(School of Basic Medicine,Guangzhou University of Chinese Medicine,Guangzhou 510006 Guangdong,China)

机构地区：[1]广州中医药大学基础医学院,广东广州510006

出　　处：《中药新药与临床药理》2018年第6期731-737,共7页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：广东省高等学校优秀青年教师培养计划项目(YQ2015042);广东省科技计划项目(2015A070706007)

摘　　要：目的探讨新橙皮苷(Neohesperidin)对五氟尿嘧啶(5-FU)化疗损伤大鼠骨髓间充质干细胞(BMSCs)的保护作用。方法全骨髓贴壁培养法原代培养BMSCs,取P3代细胞进行实验,并将实验分为正常对照组、模型组和给药组。用25μg·mL^(-1)5-FU作用于BMSCs作为模型组,用12.5μmol·L^(-1)和25μmol·L^(-1)新橙皮苷溶液分别干预25μg·mL^(-1)5-FU损伤的BMSCs细胞作为给药组,并设常规培养的BMSCs作为正常对照组。分别用CCK-8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双染流式细胞术检测细胞早期凋亡率,Western Blot法检测Caspase-3蛋白的表达水平,一氧化氮合酶(NOS)测定试剂盒检测iNOS、TNOS的含量。结果 12.5μmol·L^(-1)和25μmol·L^(-1)新橙皮苷对BMSCs无细胞毒作用且对化疗损伤的BMSCs有较好的保护作用;与正常对照组比较,模型组细胞早期凋亡率显著升高(P<0.01),Caspase-3显著上调(P<0.01),iNOS和TNOS活力明显下降(P<0.01)。与模型组比较,12.5μmol·L^(-1)和25μmol·L^(-1)给药组BMSCs细胞的早期凋亡率显著降低(P<0.05),Caspase-3显著下调(P<0.05),iNOS和TNOS活力明显上升(P<0.05)。结论新橙皮苷可能通过下调Caspase-3表达来抑制细胞凋亡,从而保护化疗损伤的BMSCs细胞。Objective To investigate the protective effects of neohesperidin on mouse bone marrow mesenchymal stem cells (BMSCs) induced by 5-fluorouracil (5-FU). Methods BMSCs were cultured by the whole bone marrow adherent culture to the logarithmic growth phase in vitro. The P3 generation cells were divided into control group, model group and experimental group. BMSCs model group were induced with 25 μg·mL^-1 5- FU. The model cells were treated with 12.5 μmol·L^-1 and 25 μmol·L^-1 neohesperidin respectively as the experimental groups. The normal BMSCs were used as control group. Cell viability was detected by CCK- 8 assay. The apoptotic morphology was detected by Hoechst 33258 staining. The rate of apoptosis was detected using flow cytometry (FCM) by Annexin VFITC and propidium iodide (PI) double labeling technique. TNOS and iNOS were detected by nitric oxide synthase (NOS) assay kit. The expression of Caspase-3 was detected by Western Blot. Results Neohesperidin at the dosages of 12.5 μmol·L^-1 and 25 μmol·L^-1 had no effect on the survival rate of BMSCs cells,and had a protective effect on BMSCs induced by chemotherapy. Compared with the control group,the early apoptosis rate of model group cells was significantly increased (P<0.01), Caspase- 3 was significantly up-regulated (P<0.01), the activities of iNOS and TNOS were significantly decreased (P<0.01). Compared with the model group,the early apoptosis rates of injury BMSCs with 12.5 μmol·L^-1 and 25 μmol·L^-1 neohesperidin treatment were significantly decreased (P< 0.05), Caspase-3 levels were significantly down- regulated (P<0.05), the activities of iNOS and TNOS weresignificantly increased (P<0.05). Conclusion The results showed that neohesperidin could protect BMSCs against 5-FU damage,which may inhibit apoptosis by down-regulating the expression of Caspase-3.

关 键 词：新橙皮苷 五氟尿嘧啶 骨髓间充质干细胞 化疗损伤 

分 类 号：R285.6[医药卫生—中药学]