摘要
目的:观察川芎嗪对氧化型低密度脂蛋白诱导人脐动脉平滑肌细胞增殖的抑制作用。方法:实验于2004/2005在泰山医学院基础医学研究所完成。①分离健康人新鲜血浆制备氧化型低密度脂蛋白。②取新鲜健康人脐带,离体培养人脐动脉平滑肌细胞。③制备含药血清:取Wistar大鼠20只,雌雄各半。随机分为川芎嗪组15只,对照组5只。川芎嗪组大鼠分别给予10,30,100mg/kg灌胃,1次/d,连续3d;对照组以等量M199灌胃,饮水不受限制。第3天给药1.5h后,从腹主动脉取血6mL,分离血清备用。④采用生长稳定的三四代人脐动脉平滑肌细胞,用无血清培养基培养24h使其生长同步化后,应用氧化型低密度脂蛋白建立平滑肌细胞增殖模型。实验分为5组,空白对照组,氧化型低密度脂蛋白组,氧化型低密度脂蛋白+10μmol/L川芎嗪组,氧化型低密度脂蛋白+30μmol/L川芎嗪组;氧化型低密度脂蛋白+100μmol/L川芎嗪组。其中,氧化型低密度脂蛋白的剂量为50μmol/L。经不同浓度川芎嗪处理72h后进行细胞计数,于川芎嗪处理12,24,72h采用氚-胸腺嘧啶核苷掺入法检测药物效应。结果:①氧化型低密度脂蛋白处理组与对照组相比,人脐动脉平滑肌细胞数目增加,氚-胸腺嘧啶核苷掺入率增加,DNA合成也增加。②与氧化型低密度脂蛋白处理组相比,氧化型低密度脂蛋白与不同浓度的川芎嗪同时处理组细胞数目减少,氚-胸腺嘧啶核苷掺入率也降低,DNA合成下降(r=0.9316,P<0.05)。③随川芎嗪浓度增加对脐动脉平滑肌细胞增殖的抑制作用递增。结论:①氧化型低密度脂蛋白有促平滑肌细胞增殖作用,川芎嗪表现为剂量依赖性地抑制平滑肌细胞计数及氚-胸腺嘧啶核苷掺入量。②可初步认为川芎嗪降血压作用可能与其抑制人脐动脉平滑肌细胞增殖有关。
AIM： To explore the inhibitive effects of tetramethylpyrazine （TMP） on oxidized low density lipoprotein （ox-LDL） induced proliferation of human umbilical artery smooth muscle cells.
METHODS： The experiment was performed at the Institute of Basic Medicine, Taishan Medical College from 2004 to 2005. ①Fresh plasma of healthy person was isolated to prepare ox-LDL.②Fresh healthy human umbilical cord was obtained and human umbilical artery smooth muscle cells were cultured in vitro. ③Preparation of drug containing serum： A total of 20 Wistar rats, male and female （half and half）, were selected and randomly assigned into TMP group with 15 rats and control group with 5 rats. Rats in the TMP group were treated with 10,30,100 mg/kg TMP by gastric perfusion, respectively, once a day for 3 days successively. Those in the control received M199 of the same volume by gastric perfusion, and drinking water was admitted. After 1.5 hours on the third day, 6 mL blood was gained from abdominal aorta and serum was isolated for preparation. ④ The well-growth third and fourth generation human umbilical artery smooth muscle cells were applied, and cultured with serum-free medium for 24 hours. After its growth was synchronizedr proliferation models were established with ox-LDL. There were 5 groups in the trial, blank control group, ox-LDL group, ox-LDL plus 10 μmol/L TMP group, ox-LDL plus 30μmol/L TMP group and ox-LDL plus 100μmol/L TMP group. The dosage of ox-LDL was 50μmol/L, After being disposed with TMP of different concentrations for 72 hours, cell counting was conducted. At hours 12, 24 and 72, the effects of TMP was determined with ^3H - thymidine（^3H-TdR） incorporation method.
RESULTS： ① Compared with control group, the number of human umbilical artery smooth muscle cells and incorporation efficiency of ^3H-TdR were increased in the ox-LDL group, besides, there was rising proliferation of DNA also in latter group.②Compared with ox-LDL group, smooth muscle cells number and incorporation efficiency of ^3H-TdR and DNA synthesis in various thickness TMP groups were lower （r= 0.931 6,P 〈 0.05）.③With the rising thickness of TMP, the inhibitive effect on proliferation of human umbilical artery smooth muscle cells was added gradually.
CONCLUSION： ①ox-LDL can promote proliferation of human umbilical artery smooth muscle cells, and the effect of TMP on smooth muscle cell counting and incorporation of ^3H-TdR is dose dependent.②The effect of TMP on blood pressure decrease is probably due to its inhibition on proliferation of smooth muscle cells.
出处
《中国临床康复》
CSCD
北大核心
2006年第43期98-100,共3页
Chinese Journal of Clinical Rehabilitation
基金
山东省卫生厅青年项目基金资助(JZ43)~~
