芦荟大黄素对动脉损伤后血管平滑肌细胞增殖的影响

目的 :探讨中药大黄的活性提取成分———芦荟大黄素对兔髂动脉损伤后离体培养的平滑肌细胞的增殖动力学的影响。方法 :纯种日本大耳白兔髂动脉内皮剥脱术后 48h ,取中膜做平滑肌细胞原代培养。指数增长期中的平滑肌细胞同步化于G0 期后 ,加入浓度梯度为 1 0 - 1 ～ 1 0 - 5g/L的芦荟大黄素于含 1 0 %胎牛血清的细胞培养液中 ,2 4h后 ,用3H TdR脉冲标记 6h ,然后测平滑肌细胞中TdR掺入量 (cpm ,每份样品每分钟脉冲数 )。同样地 ,细胞同步化于G0 期后 ,加入 2 0mg/L芦荟大黄素于含 1 0 %胎牛血清的细胞培养液中 ,作用 2 4h后 ,用流式细胞仪对细胞周期时相进行了测定。以上两实验与对照组加入等体积的细胞培养液。结果 :药物组与对照组比较 ,3H TdR的掺入量有显著性差异 (P <0 0 5 ) ,药物浓度为 1 0 - 1 ～ 1 0 - 2g/L时 ,抑制率 (对照组cpm -药物组cpm/对照组cpm× 1 0 0 % )高达 90 %以上。药物对平滑肌细胞的抑制呈剂量依赖关系 ,药物浓度越高 ,抑制作用越大。与对照组比较 ,加入芦荟大黄素后 ,G0 /G1 期细胞百分比升高 ,S期百分比下降。细胞由G0 期向S期的转化受到阻滞。结论 :芦荟大黄素是一种强平滑肌细胞抑制剂 ,这种药理作用在体内可能有利于减轻平滑肌细胞的增殖 ,从而减轻导致动脉损伤后再狭?

Effect of Aloe Emodin on Proliferation of Vascular Smooth Muscle Cells after Arterial Injury

OBJECTIVE: To study the effect of Aloe-emodin (AE), an active ingredient of Rhubarb, on the kinetics of proliferation of smooth muscular cells (SMCs) cultured in vitro after rabbit iliac arterial injury. METHODS: Forty-eight hours after de-endothelialization (balloon endothelial denudation), the iliac arteries of the Japanese white rabbits were isolated and the smooth muscle cells were cultured primarily. AE was added to culture medium containing 10% fetal calf serum (FCS). The cultures were pulse-labeled with 3H-TdR and TdR uptake into VSMC were measured and the cell cycle of the cultures were analyzed by using flow cytometer. RESULTS: Compared with control, when the concentration gradient ranged from 10(-1) to 10(-5) g/L, the amount (cpm, count per minute) of 3H-TdR uptake into SMCs has significant differences (P < 0.05) and 10(-1) and 10(-2) g/L AE showed strong inhibitory effects on TdR uptake into VSMC and the percentage of inhibition [% inhibition = (cpm without AE - cpm with AE)/cpm without AE x 100%] was more than 90%. AE displayed concentration dependent inhibitory effects. The percentage of cells in G0/G1 phase was increased, but the percentage of cells in S phase was decreased in AE group, the transition of SMC cycle phase from G0 to S was blocked. CONCLUSIONS: AE is a strong inhibitor to the proliferation of SMCs and the pharmacological action of AE might reduce SMC proliferation in vivo and decrease intimal hyperplasia of restenosis.