川芎嗪对长春新碱诱导的骨髓基质细胞生长抑制和凋亡的干预  

Effect of Ligustrazine Hydrochloride on suppression and apoptosis of bone marrow stromal cells induced by Vincristine

作　　者：文珠[1] 胡国柱[2] 何丹[2] 俞火[1] 戎吉平[1] 

机构地区：[1]江西省医学科学研究所血液学研究室,南昌330006 [2]江西省人民医院临床医学研究所干细胞研究室,南昌330006

出　　处：《中华中医药杂志》2010年第12期2176-2179,共4页China Journal of Traditional Chinese Medicine and Pharmacy

摘　　要：目的:探讨川芎嗪(ligustrazine hydrochloride,LH)对小鼠骨髓基质细胞(BMSCs)生长的促进及干预长春新碱(VCR)诱导的凋亡作用。方法:①LH(5、10、20、40μg/mL)及VCR(2.5、5、10、15μg/mL)分别与BMSCs培养14d,应用MTT法测定细胞增殖;②不同浓度LH与BMSCs培养1h后再加入5μg/mL VCR继续培养14d,测定细胞增殖;③BMSCs培养14d后再加入不同浓度VCR继续培养24h,应用Hoechst33342荧光染色及Annexin V/PI双染流式细胞术(FACS)方法测定细胞凋亡。④不同浓度LH与BMSCs培养14d后再加入5μg/mL VCR继续培养24h测定细胞凋亡。结果:①随LH的浓度增加BMSCs增殖率升高,其中20μg/mL和40μg/mL两组OD值与正常对照组比较差异显著(P<0.05)。而VCR组随浓度增加BMSCs增殖率下降,10μg/mL和15μg/mL两组OD值显著低于正常对照组(P<0.05)。②BMSsC与LH孵育1h后再加入5μg/mL VCR,培养14d后10μg/mL、20μg/mL、40μg/mLLH三组OD值与单用5μg/mLVCR组比较差异均有显著性(P<0.05)。③BMSCs培养14d再加VCR继续培养24h后FACS测定5μg/mL、10μg/mL、15μg/mL VCR三组凋亡率与正常对照组比较均有显著性差异(P<0.05)。④LH与BMSCs培养14d再加入5μg/mL VCR继续培养24h后,FACS测定20μg/mL和40μg/mL LH两组凋亡率显著低于凋亡诱导组(P<0.05);Hoechst33342检测,则10μg/mL、20μg/mL、40μg/mL LH3组凋亡率依次为(30.67±8.02)%、(20.33±4.16)%、(19.0±4.58)%,显著低于VCR5μg/mL凋亡诱导组(47.0±6.00)%(P<0.05)。结论:川芎嗪可促进BMSCs增殖,干预长春新碱对BMSCs生长的抑制;川芎嗪能干预长春新碱诱导的BMSCs凋亡。Objective：To investigate the effect of LH（Ligustrazine Hydrochloride Injection）on the proliferation promotion and apoptosis inhibition of mice bone marrow stromal cells（BMSCs）induced by VCR（Vincristine）.Methods：The BMSCs were cultured with LH（5,10,20,40μg/mL）and VCR（2.5,5,10,15μg/mL）in 37℃5%CO2 for 14 days,respectively and the BMSCs proliferation were determined by MTT.The BMSCs were pretreated by LH（5,10,20,40μg/ml）for 1hour,then cultured with VCR 5μg/ml in 37℃5%CO2 for 14 days,and the BMSCs proliferation were determined by MTT.The BMSCs were cultured for 14 days,then VCR（2.5,5,10,15μg/ml）was added to culture in 37℃5%CO2 for 24hours,respectively,and cell apoptosis was determined by FACS with Annexin V/PI.The BMSCs were cultured with LH（5,10,20,40μg/mL）for 14 days,then 5μg/ml VCR was added to culture in 37℃5%CO2 for 24 hours,respectively,and cell apoptosis was determined by Hoechst 33342 fluorescent staining and FACS with Annexin V/PI.Results：The BMSCs grew more quick along with LH concentration increasing,and the OD value was significantly higher in the LH groups of 20μg/ml and 40μg/ml than in the normal control group on 14th day（P0.05）.In the VCR groups the BMSCs grew slower along with VCR concentration increasing,and the OD value was significantly slower in the VCR groups of 10μg/ml and 15μg/ml than in the normal control group in 14th day（P0.05）.The BMSCs were pretreated with LH for 1 hour,then cultured with 5μg/ml VCR for 14 days,and proliferation of the BMSCs increased along with LH concentration increasing.In the LH groups of 10μg/ml,20μg/ml,40μg/ml the OD value was significantly higher than in the only VCR 5μg/ml group（P0.05）.After the BMSCs were cultured for 14 days,VCR（2.5,5,10 or 15μg/ml）was added to culture and continued to be cultured for 24 hours,then the apoptosis rate of BMSCs detected by FACS with Annexin V/PI was significantly higher in the VCR groups of 5μg/ml,10μg/ml,15μg/ml than in the normal control group（P0.0

关 键 词：川芎嗪 长春新碱 骨髓基质细胞 增殖 凋亡 

分 类 号：R285[医药卫生—中药学]