鳖甲煎丸对体外培养大鼠主动脉环血管新生的影响

The Effect of Biejia Jian Wan (鳖甲煎丸) on Angiogenesis of vitro Cultured Aorta Ring in Rats

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 Biejia Jian Wan（鳖甲煎丸） medicine-containing serum of different concentration on angiogenesis. Methods Randomly divided 14 SD rats into the Biejia Jian Wan group（12 rats）and the saline group（2 rats）. The Biejia Jian Wan group was given Biejia Jian Wan 6 mg /（ kg·d） for gavage while the saline group was given the same volume of saline for gavage,both groups for 5 days. Took blood from abdominal aorta and separated serum one hour after the last gavage. Adopted in vitro cultured rat abdominal aorta ring to develop the model. Used culture medium contending Biejia Jian Wan serum of different concentrations（ containing 5%,10%,15%,20%,30% and 40% respectively） for culture. Set the control group（ serum of the saline group）,cultivated for 10 days in total. Observed and filmed microvascular growth images under inverted microscope. Calculated microvascular budding areas; Conducted immunohistochemical staining on the culture. Calculated the average density of microvascular in every high power field. Conducted Spearman ranking correlation analysis between density of nascent capillaries in aorta ring and the concentration of Biejia Jian Wan serum. Results Compared with the control group,microvascular budding areas of the 20%,15%,10% and 5% Biejia Jian Wan serum groups all remarkably decreased（ P〈0. 01）. There is no statistical significance in difference between microvascular budding areas of the30% and 40% Biejia Jian Wan serum groups（ P〈0. 05）. Microvascular budding area of the 15% Biejia Jian Wan serum group was the lowest,remarkably lower than the 5%,20%,30%,and 40% Biejia Jian Wan serum group（P〈0. 01）. But the difference between it and the 10% Biejia Jian Wan serum group has no statistical significance（P〈0. 05）. As is shown by related analysis results,when the concentration of Biejia Jian Wan serum≤15%,its function to inhibit angiogenesis in aorta ring generally reinforced and density of nascent capillaries in aorta ring decreased with concentration increased（ r =- 0. 637,P〈0.. 05） Conclusion Biejia Jian Wan had restraining function on new vessels and drug concentration of 15% seems to be the most effective.