平肝潜阳方含药血清中天麻素水平测定及其对血管平滑肌细胞活性的影响  

Determination of gastrodin level in serum containing formula for calming the liver and suppressing yang and investigation of drug serum effects on vascular smooth muscle cell avtivity in rats

作　　者：李湘平[1] 罗艳红[2] 钟广伟[3] 尹桃[1] 相玲丽[3] 李炜[3] 陈国林[3] 

机构地区：[1]中南大学湘雅医院药剂科,湖南省长沙市410008 [2]中南大学湘雅医院急诊科,湖南省长沙市410008 [3]中南大学湘雅医院中西医结合研究所,湖南省长沙市410008

出　　处：《中国组织工程研究与临床康复》2010年第15期2735-2739,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家自然科学基金资助项目(30500644):运用差异蛋白质组技术研究平肝潜阳药治疗肝阳上抗证的分子机制;湖南省中医药科研计划项目(2009047):从MicroRNA角度研究平肝潜阳方逆转高血压血管重构的作用机制~~

摘　　要：背景:平肝潜阳方为中药复方制剂,化学成分复杂,直接测定血清中药物含量,比较困难。天麻是平肝潜阳方的君药之一,且其为成苷类衍生物,水平容易测定。目的:通过检测血清中天麻素水平了解平肝潜阳方在体内的药物浓度,探讨制备含药血清的具体方法,并观察平肝潜阳方含药血清对血管平滑肌细胞活性的影响。方法:大鼠予以平肝潜阳方浸膏2g/d给药3d或7d,分别于3d或7d内于每天8:00和16:00各灌药1g,与第4或第8天8:00灌药2g,于最后给药前及给药后0.5,1,2,3,5h心脏内采血。高效液相色谱测定大鼠体内天麻素水平;在血管平滑肌细胞的培养液中分别添加不同浓度灭活处理的含药血清,加药培养后24h采用MTT法观察细胞活性的抑制率。结果与结论:平肝潜阳方浸膏3d与7d给药大鼠,均在最后一次给药后1h达到最高血药浓度,但3d给药血药浓度迅速下降,而7d给药可维持较长时间的血药浓度平台;天麻素在0.045～5.6mg/L范围内呈线性,r=0.9992,精密度RSD日内为1.86%,加样回收率为93.88%。不同浓度含药血清对血管平滑肌细胞的生长均有抑制作用,但以5%浓度含药血清对血管平滑肌细胞的生长抑制率最大(P<0.01),其中7d给药组的抑制率明显高于3d给药组(P<0.01)。因此,平肝潜阳方含药血清的制备应选取连续给药7d后末次给药1h后为最佳采血时间点,含药血清能够抑制血管平滑肌细胞的活性。BACKGROUND：The formula for calming the liver and suppressing yang is a Chinese herbal medicine compound preparation composed complex chemical constituents,thus,it is difficult to determine its serum levels.However,as a principal drug for this formula,the level of gastrodin is easy to measure.OBJECTIVE：With a method of serum-pharmacology,to explore the prepare method of serum containing formula for calming the liver and suppressing yang,and to observe its effects on the activity of vascular smooth muscle cells（VSMCs） in rats.METHODS：Rats were given 1g formula for calming the liver and suppressing yang extract at 8：00 and 16：00 continuous for 3 days or 7 days,respectively.Then additional 2 g drugs were administrated at 8：00 in the next days of 3 or 7 days.The blood was taken from heart at prior to and 0.5,1,2,3 and 5 hours after the last administration.Gastrodin levels in these serums were determined by high performance liquid chromatogram.Subsequently,inactivated serum containing formulae for calming the liver and suppressing yang with different concentrations was used to culture VSMCs,and the inhibitory rates of drug serum on VSMCs activity were observed at 24 hours after culture using MTT assay.RESULTS AND CONCLUSION：Concentration of gastrodin reached the highest level at 1 hour after last administration of formulae for calming the liver and suppressing yang in the 3-day administration or 7-day administration groups.However,the gastrodin level was rapidly decreased in the 3-day administration group,which maintained for a long time in the 7-day administration group.A linearity was obtained over the concentration of 0.045-5.6 mg/L,r=0.999 2.The average recovery was 93.88% with relative standard deviation（RSD）within day=1.86%.Serum containing each herbal medicine could inhibit VSMCs activity at different concentrations.The greatest growth inhibitory ratio of VSMCs was obtained using 5.0% concentration serum containing Chinese herbal medicine（P〈0.01）,and compared with the 3-day administra

关 键 词：平肝潜阳方 天麻素 中药血清 高效液相色谱 血管平滑肌细胞 

分 类 号：R318[医药卫生—生物医学工程]