【作者】 文珠； 胡国柱； 何丹； 俞火； 戎吉平；

【Author】 Wen Zhu~1,Hu Guo-zhu~2,He Dan~2,Yu Huo~1,Rong Ji-ping~1 (1.Jiangxi Institute of Medical Science,Nanchang University,Nanchang 330006,P.R.China;2. Institute of Clinical Medicine of The Jiangxi Province People’s Hospital,Nanchang,330006, P.R.China)

【机构】 江西省医学科学研究所； 江西省人民医院临床医学研究所；

【摘要】 目的:探讨川芎嗪(Ligustrazine Hydrochloride,LH)对小鼠骨髓基质细胞(BMSC)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BMSC培养14d,应用MTT法测定细胞增殖;②不同浓度LH与BMSC培养1h后再加入5μg/ml VCR继续培养14d,测定细胞增殖;③BMSC培养14d后再加入不同浓度VCR继续培养24h,应用Hoechst 33342荧光染色及AnnexinV/PI双染流式细胞术(FACS)方法测定细胞凋亡。④不同浓度LH与BMSC培养14d后再加入5μg/mlVCR继续培养24h测定细胞凋亡。结果:①随LH的浓度增加细胞增殖率升高,第14d 20μg/ml(1.97)和40μg/ml(1.98)LH两组OD值与正常对照组(1.53)比较差异有显著性(P<0.05)。而VCR组随浓度增加增殖率下降,第14d 10μg/ml(0.83)和15μg/ml(0.72)VCR两组OD值显著低于正常对照组(1.53)(P<0.05)。②BMSC与LH孵育1h后再加入5μg/ml VCR,培养14d后10μg/ml(1.56)、20μg/ml(1.83)、40μg/ml(1.77)LH三组OD值与单用5μg/ml VCR(1.07)比较差异均有显著性(P<0.05)。③BMSC培养14d再加VCR继续培养24h后FACS测定5μg/ml、10μg/ml、15μg/ml三组凋亡率分别为31.12%、39.37%、64.10%,与正常对照组(7.70%)比较差异有显著性(P<0.05)。④LH与BMSC培养14d再加入5μg/ml VCR继续培养24h后,FACS测定20μg/ml和40μg/ml两组凋亡率为13.0%和12.71%,显著低于凋亡诱导组(31.12%)(P<0.05);Hoechst 33342检测,则10μg/ml、20μg/ml、40μg/ml LH三组凋亡率依次为30.67%、20.33%、19.0%,显著低于VCR 5μg/ml凋亡诱导组(47.0%)(P<0.05)。结论:川芎嗪可促进骨髓基质细胞增殖,干预长春新碱对骨髓基质细胞生长的抑制。川芎嗪能干预长春新碱诱导的骨髓基质细胞凋亡。更多还原

【Abstract】 Objective:To investigate the effect of LH（Ligustrazine Hydrochloride for Injection） on the proliferation promotion and apoptosis inhibition of mice bone marrow stromal cells（BMSCs） induced by VCR（Vincristine）.Methods:Ⅰ.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.Ⅱ.The BMSCs were pretreated by LH（5,10,20, 40μg/ml） for 1hr,then cultured with VCR 5μg/ml in 37C 5%CO₂ for 14 days,and the BMSCs proliferation were determined by MTT.Ⅲ.The BMSCs were cultured for 14 days,then VCR（2.5, 5,10,15μg/ml） was added to culture in 37℃5%CO₂ for 24 hours,respectively,and cell apoptosis was determined by FACS with Annexin V/PI.Ⅳ.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（1.97） and 40μg/ml（1.98） than in the normal control group（1.53） in 14th day（P<0.05）.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（0.83） and 15μg/ml（0.72） than in the normal control group（1.53） in 14th day（P<0.05）.The BMSCs were pretreated with LH for 1h,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（1.56）,20μg/ml（1.83）,40μg/ml（1.77） the OD value was significantly higher than in the only VCR 5μg/ml（1.07） group（P<0.05）.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（31.12%）,10μg/ml（39.37%）, 15μg/ml（64.10%） than in the normal control group（7.70%）（P<0.05）.After LH culturing with BMSCs for 14 days,5μg/ml VCR was added to culture to continue for 24 hours,then apoptosis rate of BMSCs detected by FACS with Annexin V/PI was significantly lower in the LH groups of 20μg/ml（13.0%） and 40μg/ml（12.71%） than in the only VCR 5μg/ml group（31.12%）（P<0.05）; and apoptosis rate of BMSCs detected by Hoechst 33342 fluorescent staining was significantly lower in the LH groups of 10μg/ml（30.67%）,20μg/ml（20.33%） and 40μg/ml（19.0%） than in the only VCR 5μg/ml group（47.0%）（P<0.05）.Conclusion:The results suggest that Ligustrazine can promote the BMSCs proliferation and prevent from inhibition of the proliferation and apoptosis of the BMSCs induced by vincristine.更多还原