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目的观察不同浓度鳖甲煎丸含药血清对血管新生的影响。方法 14只SD大鼠随机分成鳖甲煎丸组(12只)和生理盐水组(2只)。鳖甲煎丸组给予鳖甲煎丸6 mg/(kg·d)灌胃,生理盐水组给予等量生理盐水灌胃,连续5天。末次给药1 h后,腹主动脉取血分离血清。采用体外培养大鼠腹主动脉环血管生成模型,用含不同浓度的鳖甲煎丸血清(含5%、10%、15%、20%、30%、40%)培养基培养,并设对照组(生理盐水组血清),共培养10天。倒置显微镜下观察并拍摄微血管生长图像,计算微血管出芽面积;将培养物进行免疫组织化学染色,计算每高倍视野中平均微血管密度。并将血管环新生微血管密度与鳖甲煎丸含药血清浓度进行Spearman等级相关分析。结果与对照组比较,20%、15%、10%、5%鳖甲煎丸含药血清组微血管出芽面积均明显减少(P<0.01),30%、40%鳖甲煎丸含药血清组微血管出芽面积差异无统计学意义(P>0.05)。15%鳖甲煎丸含药血清组出芽面积最少,明显低于5%、20%、30%、40%鳖甲煎丸含药血清组(P<0.01),但与10%鳖甲煎丸含药血清组比较差异无统计学意义(P>0.05)。相关分析结果显示,鳖甲煎丸含药血清浓度≤15%时,随着浓度增加,对血管环新生血管抑制作用逐渐增强,血管环新生微血管密度逐渐减少(r=-0.637,P<0.05)。结论鳖甲煎丸对新生血管具有抑制作用,且以15%浓度含药血清效果最好。
Objective To observe the effect of different concentrations of biejiajian pill containing serum on angiogenesis. Methods Fourteen SD rats were randomly divided into Biejiaocao pill group (12 rats) and saline group (2 rats). Biejiaofu Pill group given BJJ 6 mg / (kg · d) gavage, normal saline group given the same amount of saline for 5 consecutive days. 1 h after the last administration, blood was collected from the abdominal aorta to separate the serum. The model of angiogenesis in rat abdominal aorta was cultured in vitro and cultured in different concentrations of BMRA serum (containing 5%, 10%, 15%, 20%, 30%, 40%) medium and control Group (saline group serum), co-cultured for 10 days. Under inverted microscope, the microvascular growth images were observed and photographed to calculate the microvascular sprouting area; the culture was subjected to immunohistochemical staining to calculate the average microvessel density per high power field. Spearman rank correlation analysis was performed between the neovascular microvessel density and the serum concentration of BJJ pill. Results Compared with the control group, the area of microvessel sprouting in 20%, 15%, 10%, 5% BMRA-containing serum was significantly decreased (P <0.01), and 30%, 40% There was no significant difference in the area of microvascular budding (P> 0.05). Bupijian pills containing 15% BJJ group had the lowest sprouting area, which was significantly lower than that of 5%, 20%, 30%, 40% BJJ pills (P <0.01) There was no significant difference between the drug-containing serum groups (P> 0.05). Correlation analysis showed that when the serum concentration of BJJ pill was less than or equal to 15%, the inhibitory effect on angiogenic neovascularization gradually increased with the increase of concentration, and the density of neovascularization of vascular ring gradually decreased (r = -0.637, P <0.05) . Conclusion Biejiajian Pills has an inhibitory effect on neovascularization, and the effect of containing 15% serum is the best.