中国组织工程研究与临床康复
中國組織工程研究與臨床康複
중국조직공정연구여림상강복
JOURNAL OF CLINICAL REHABILITATIVE TISSUE ENGINEERING RESEARCH
2010年
15期
2735-2739
,共5页
李湘平%罗艳红%钟广伟%尹桃%相玲丽%李炜%陈国林
李湘平%囉豔紅%鐘廣偉%尹桃%相玲麗%李煒%陳國林
리상평%라염홍%종엄위%윤도%상령려%리위%진국림
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
平肝潛暘方%天痳素%中藥血清%高效液相色譜%血管平滑肌細胞
평간잠양방%천마소%중약혈청%고효액상색보%혈관평활기세포
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2 g/d给药3 d或7 d,分别于3 d或7 d内于每天8:00和16:00各灌药1 g,与第4或第8天8:00灌药2 g,于最后给药前及给药后0.5,1,2,3,5 h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24 h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3 d与7 d给药大鼠,均在最后一次给药后1 h达到最高血药浓度,但3 d给药血药浓度迅速下降,而7 d给药可维持较长时间的血药浓度平台;天麻素在0.045-5.6 mg/L范围内呈线性,r=0.999 2,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7 d给药组的抑制率明显高于3 d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7 d后末次给药1 h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.
揹景:平肝潛暘方為中藥複方製劑,化學成分複雜,直接測定血清中藥物含量,比較睏難.天痳是平肝潛暘方的君藥之一,且其為成苷類衍生物,水平容易測定.目的:通過檢測血清中天痳素水平瞭解平肝潛暘方在體內的藥物濃度,探討製備含藥血清的具體方法,併觀察平肝潛暘方含藥血清對血管平滑肌細胞活性的影響.方法:大鼠予以平肝潛暘方浸膏2 g/d給藥3 d或7 d,分彆于3 d或7 d內于每天8:00和16:00各灌藥1 g,與第4或第8天8:00灌藥2 g,于最後給藥前及給藥後0.5,1,2,3,5 h心髒內採血.高效液相色譜測定大鼠體內天痳素水平;在血管平滑肌細胞的培養液中分彆添加不同濃度滅活處理的含藥血清,加藥培養後24 h採用MTT法觀察細胞活性的抑製率.結果與結論:平肝潛暘方浸膏3 d與7 d給藥大鼠,均在最後一次給藥後1 h達到最高血藥濃度,但3 d給藥血藥濃度迅速下降,而7 d給藥可維持較長時間的血藥濃度平檯;天痳素在0.045-5.6 mg/L範圍內呈線性,r=0.999 2,精密度RSD日內為1.86%,加樣迴收率為93.88%.不同濃度含藥血清對血管平滑肌細胞的生長均有抑製作用,但以5%濃度含藥血清對血管平滑肌細胞的生長抑製率最大(P<0.01),其中7 d給藥組的抑製率明顯高于3 d給藥組(P<0.01).因此,平肝潛暘方含藥血清的製備應選取連續給藥7 d後末次給藥1 h後為最佳採血時間點,含藥血清能夠抑製血管平滑肌細胞的活性.
배경:평간잠양방위중약복방제제,화학성분복잡,직접측정혈청중약물함량,비교곤난.천마시평간잠양방적군약지일,차기위성감류연생물,수평용역측정.목적:통과검측혈청중천마소수평료해평간잠양방재체내적약물농도,탐토제비함약혈청적구체방법,병관찰평간잠양방함약혈청대혈관평활기세포활성적영향.방법:대서여이평간잠양방침고2 g/d급약3 d혹7 d,분별우3 d혹7 d내우매천8:00화16:00각관약1 g,여제4혹제8천8:00관약2 g,우최후급약전급급약후0.5,1,2,3,5 h심장내채혈.고효액상색보측정대서체내천마소수평;재혈관평활기세포적배양액중분별첨가불동농도멸활처리적함약혈청,가약배양후24 h채용MTT법관찰세포활성적억제솔.결과여결론:평간잠양방침고3 d여7 d급약대서,균재최후일차급약후1 h체도최고혈약농도,단3 d급약혈약농도신속하강,이7 d급약가유지교장시간적혈약농도평태;천마소재0.045-5.6 mg/L범위내정선성,r=0.999 2,정밀도RSD일내위1.86%,가양회수솔위93.88%.불동농도함약혈청대혈관평활기세포적생장균유억제작용,단이5%농도함약혈청대혈관평활기세포적생장억제솔최대(P<0.01),기중7 d급약조적억제솔명현고우3 d급약조(P<0.01).인차,평간잠양방함약혈청적제비응선취련속급약7 d후말차급약1 h후위최가채혈시간점,함약혈청능구억제혈관평활기세포적활성.
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 2g 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=0.9992.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 administration group,the VSMCs of the 7-day group exhibited remarkable growth inhibition ratio(P<0.01)Accordingly,the optimal time for blood collecting is at 1 hour after a 7-day successive administration.The serum containing formula for calming the liver and suppressing yang can inhibit the growth of VSMCs.