【摘要】 背景:中医活血化瘀治法是针对血瘀证而设立的,目前关于活血化瘀治疗对干细胞动员和归巢作用的研究甚少。目的:观察丹参、血府逐瘀汤含药血清对兔骨髓间充质干细胞体外迁移的影响。设计、时间及地点:细胞学体外观察,于2007-06/12在南昌大学心血管病研究所完成。材料:健康CV级6～8周龄雄性新西兰大耳兔24只,随机分为对照组、丹参组、血府组,8只/组。另取二三周龄雄性新西兰大耳兔24只用于制备骨髓间充质干细胞。丹参饮片、血府逐瘀汤饮片由江西省中医院临床药物实验基地提供。方法:取丹参及血府逐瘀汤的饮片,中药文火水煎二次过滤去渣,合并水煎液后浓缩成含生药0.5g/mL溶液。丹参组灌服丹参溶液2.5g/(kg?d),1次/d,连续7d;血府组同法灌服血府逐瘀汤3.95g/(kg?d);对照组灌服等量蒸馏水。分别于药物干预前、干预7d后抽取各组兔动脉血分离制备相应血清。取transwell系统,下室加入各组对应血清,上室加入第3代细胞骨髓间充质干细胞悬液,于37℃、体积分数为5%的CO2饱和湿度培养箱中培养。主要观察指标:ELISA法测定血清间质细胞衍生因子1的质量浓度,利用transwell系统观察含药血清对兔骨髓间充质干细胞迁移的影响,MTT法测定骨髓间充质干细胞的倍增时间。结果:①干预前,各组血清间质细胞衍生因子1的质量浓度大致相同;干预7d后,对照组间质细胞衍生因子1质量浓度无明显改变(P>0.05),丹参组、血府组间质细胞衍生因子1质量浓度均明显升高(P<0.05)。②干预12h后,对照组迁移细胞数增加1倍。与对照组比较,干预4h后丹参组、血府组骨髓间充质干细胞迁移数均明显增加(P<0.05),此后一直呈增加趋势。③与对照组比较,丹参组、血府组骨髓间充质干细胞的倍增时间均明显缩短(P<0.05)。结论:丹参和血府逐瘀汤含药血清均能促进兔骨髓间充质干细胞的体外迁移和增殖,且效果相当,其作用机制可能与升高血清间质细胞衍生因子1的质量浓度有直接关系。更多还原

【Abstract】 BACKGROUND:Chinese medical science of xuefuzhuyu method is used to treat blood stasis syndrome;however,studies regarding xuefuzhuyu method on stem cell mobilization and homing are poorly understood. OBJECTIVE:To explore the effects of salvia miltiorrhiza and xuefuzhuyu decoction on the migration of rabbit bone-derived mesenchymal stem cells(BMSCs) . DESIGN,TIME AND SETTING:An in vitro cytology observation. The experiment was performed at the Institute of Cardiovascular Research,Nanchang University between June and December 2007. MATERIALS:Twenty-four adult,healthy,CV grade,New Zealand rabbits,aged 6-8 weeks,were randomly divided into control,salvia miltiorrhiza,and xuefuzhuyu groups,with 8 animals in each group. Additional 24 rabbits with 2-3 weeks were collected to prepare BMSCs. salvia miltiorrhiza crude slices and xuefuzhuyu decoction were supplied by clinical drug base of Jiangxi Provincial College of Traditional Chinese Medicine. METHODS:The salvia miltiorrhiza crude slices and xuefuzhuyu soup were decocted with water using mild fire,removed dregs twice,followed by mixed the liquid to harvest solution containing 0.5 g/mL crude drug. Rabbits in the salvia miltiorrhiza group were drench salvia miltiorrhiza solution 2.5g/(kg?d) ,once per day for 7 successive days. Totally 3.95g/(kg?d) xuefuzhuyu decoction or same volume distilled water were drench in the xuefuzhuyu and control groups,respectively. The serum was collected at 7 day prior to and after intervention. subsequently,the serum and the 3rd passage BMSCs suspension were cultured in a humidified atmosphere of 5% CO2 at 37 ℃ using transwell system. MAIN OUTCOME MEASURES:The stromal derived factor-1(SDF-1) concentration in serum was measured by ELISA method;the effect of medicated serum on the migration of BMSCs was evaluated with transwell system;in addition,the doubling generation time of BMSCs was detected by MTT. RESULTS:Before the intervention,the serum SDF-1 levels were approximately identical in 3 groups,however,it had no changes in the control group at 7 days after intervention(P > 0.05) ,but,the serum SDF-1 levels were obvious increased in the salvia miltiorrhiza and xuefuzhuyu groups(P < 0.05) . The number of migrated BMSCs in the control group increased 1 fold at 12 hours after intervention. Compared to the control group,the migrated BMSCs numbers were significantly higher in the salvia miltiorrhiza and Xuefuzhuyu groups at 4 hour after intervention(P < 0.05) ,which showed an increasing trend. Simultaneously,the doubling generation time of BMSCs in the salvia miltiorrhiza and Xuefuzhuyu groups were significantly shorter than in that of the control group(P﹤0.05) . CONCLUSION:The serum treated with salvia miltiorrhiza and xuefuzhuyu soup has equivalent promoting effect on migration and proliferation of BMSCs,which may be directly related to the increased SDF-1 level.更多还原