【作者】 李征；

【导师】 何剪太；

【作者基本信息】 中南大学 ， 生物医学工程， 2010， 硕士

【摘要】 实验目的：观察姜黄素对血管平滑肌细胞增殖的抑制作用;探讨其作用机理。方法：(1)体内实验：将实验大鼠分为4组：假手术组(N组)、模型组(M组)、高剂量药物作用组(H组)、低剂量药物作用组(L组)。行P53,Fas组化检测;Bcl-2、Bax免疫印迹检测；TUNEL染色。结果用麦克奥迪图象分析系统分析,计算每张片子的平均光密度,药物用组分别与模型组进行比较,行单因素方差分析,计算P值。(2)体外实验：将VSMC随机分为五组,分别为空白组、模型组(AngⅡ10-7mol/L)和模型+姜黄素1、2、3组(姜黄素剂量为10、20、40、80μmol/L),各组VSMC用相应的细胞培养液孵育48小时,分别应用MTT检测生长率,用流式细胞术检测VSMC的细胞周期构成比,用TUNEL检测凋亡率。实验数据用平均数±标准差(X±S)表示,各组之间的数值比较采用样本均数方差分析进行统计学分析。结果：(1)体内实验：高剂量药物作用组和假手术组Fas、P53的表达低于模型组,P<0.05;低剂量药物作用组与模型组相比无显著性差异,P>0.05。高、低剂量药物作用组和假手术组的Bcl-2和TUNEL的农达低于模型组,P<0.05。高、低剂量药物作用组和假手术组Bax表达高于模型组,P<0.05。(2)姜黄素作用后的VSMC较模型组的VSMC都有增殖减少,并与姜黄素的浓度呈反比；姜黄素作用后的VSMCG0/G1期构成比较模型组的VSMC升高,与姜黄素的浓度呈正比,S期、G2/M期构成比较模型组的VSMC降低,增殖指数下降,S期与姜黄素的浓度呈反比,G2/M期与姜黄素的浓度无相关；姜黄素作用后的VSMC较模型组的VSMC凋亡率增加,并与姜黄素的浓度呈正比。结论：姜黄素能抑制体内与体外平滑肌细胞Fas、P53表达;影响Bcl-2、Bax基因表达,促进血管平滑肌细胞凋亡;达到制平滑肌细胞增殖的作用。更多还原

【Abstract】 Objeetive:To explore the influence of CUR on vascular smooth muscle cell proliferation and its mechanism. Methods:(1)In vivo test: Wistar rats were divided into 4 groups:big dose drug group, small dose drug group, mode group, false operation group.P53, Fas were detected by histoehemistry and were detected in Bcl-2. Bax by West Blot and TUNEL tests in these paraffin slices.The average light density of every slice were detected by computer image analysis system. The results were analyzed by one-way analysis of variance.And then the results of every group were compared with those of mode group by using Dunnet-t test. (2) In vitro test:Divide VSMC into 5 groups at random.The groups are respectively named with control group,modal group(AngⅡ10-/mol/L),modal+ Curcumin groups,among the model+ Curcumin groups, Curcumin is divided to 10、20、40、80μmol/L different density. Every group is cultured 48 hours.To determine the cell proliferation by MTT,to detect the cell cycle by flow cytometry,to detect the apoptosis by TUNEL.The experimental data was denoted to mean±standard deviation(x±s),use analysis of variance to statistics analyze the numerus compare of each group.Results:(1) In vivo test:The expression of P53 and Fas in slices of three groups are lower than the mode group.Compared with P53 and Fas results of mode group there is significant difference from the results of false operation group and big dose drug group P<0.05. Compared with P53 and Fas results of mode group there is not significant difference from the results of small dose drug group P>0.05.The expression of Bcl2 in slices of three groups are lower than the mode group.Compared with Bcl2 and TUNEL test results of mode group there is significant difference from the results of false operation group, small and big dose drug group P<0.05. The expression of Bax in slices of three groups are higher than the mode group.Compared with Bax results of mode group there is significant difference from the results of false operation group, small and big dose drug group P<0.05..(2)In vitro test:The VSMC proliferation rate after Curcumin is lower than that after AngⅡ. It shows inverse proportion to the density of Curcumin. Constituent ratio of VSMC G0/G1 cell cycle after Curcumin is higher thanthat after AngⅡ. It shows direct proportion to the density of hebesser. Constituent ratio of VSMC S cell cycle、G2/M cell cycle and proliferation index after Curcumin is lower than those after AngⅡ.The S cell cycle shows inverse proportion to the density of Curcumin,the G2/M cell cycle doesn’t relate to the density of Curcumin.The VSMC apoptosis rate after Curcumin is higher than that after AngⅡ.It shows direct proportion to the density of Curcumin.CONCLUSION The results show that Curcumin can markedly inhibit expression of P53, Fas in vascular smooth muscle cells, adjust expression of Bax,Bcl2 gene relating with VSMC apoptosis and pomote VSMC apoptosis. Curcumin can Inhibit VSMC Proliferation.更多还原