乐脉颗粒促进鸡胚绒毛尿囊膜血管的生成

Angiogenetic effect of Le-Mai granule on the chick embryo chorioallantoic membrane

目的:乐脉颗粒在缺血性心脑血管病的治疗中具有较好的效果,其是否通过促进血管生成而发挥作用还不清楚。观察乐脉颗粒对鸡胚绒毛尿囊膜血管生成的影响。方法:实验于2005-03/08在江西省重点实验室南昌大学第二附属医院分子中心完成。①实验材料:新西兰大白兔8只,体质量2～2.5kg;新鲜白皮种蛋70只,质量50～60g。②实验方法:种蛋在(37.5±0.5)℃条件下孵育,种蛋受精率90%以上。第7天开窗暴露鸡胚绒毛尿囊膜建立鸡胚绒毛尿囊膜模型。将60枚存活鸡胚随机分为生理盐水组、正常血清组、乐脉血清组、内膜损伤血清组、内膜损伤乐脉治疗组以及血管内皮生长因子(20mg/L)组,每组10枚鸡胚。新西兰大白兔腹主动脉用球囊导管损伤建立血管内膜损伤模型,乐脉血清组及内膜损伤乐脉治疗组给予乐脉颗粒25mg/(kg·d)喂饲,各组7d后取血清。第8天,在鸡胚绒毛尿囊膜上放一直径为5mm的滤膜作为载体,分别加样5μL,1次/d,连续3d,③实验评估:第11天取鸡胚绒毛尿囊膜,数码相机拍照后平铺于载玻片上,计数载体周围血管数目及滤膜周围0.5cm范围内的血管分支点数并进行比较。结果:纳入大白兔8只,存活鸡胚60枚,均进入结果分析,无脱落。与正常血清组相比较,乐脉血清组鸡胚绒毛尿囊膜周围血管总数明显增多,血管以载体为中心呈辐辏状生长,差异具有显著性意义(P<0.05);与正常血清组相比较,内膜损伤血清组血清载体周围血管数量明显增多,差异具有极显著性意义(P<0.01);与内膜损伤乐脉治疗组相比较,血管总数差异无显著性。结论:①兔乐脉颗粒血清能够明显促进鸡胚绒毛尿囊膜上血管生成。②血管内膜损伤7d后的血清能够促进鸡胚绒毛尿囊膜上血管生成。

AIM： Le-Maigranule is effective in treatment of ischemic cardio-cerebrovascular diseases, but its angiogenesis effect is unclear. This study is designed to investigate the angiogenesis effect of Le-Mai granule on the chick embryo chorioallantoic membrane （CAM） model. METHODS： From March to August in 2005, the experiment was carded out in the Jiangxi Provincial Key Laboratory, Molecule Center at the Second Affiliated Hospital of Nanchang University, ①Expedmental materials： Eight New Zealand rabbits weighed 2-2,5 kg; Seventy fresh white hatching eggs weighed 50-60 g,②Experimental methods： Hatching eggs were incubated at （37.5±0.5℃ and fertilized for over 90%. Every egg underwent fenestration on day 7 to expose CAM and establish CAM model. Sixty survival chick embryos were randomly divided into 6 groups （n =10）： normal saline group, normal serum group, Le-Mai treatment group （T）, intimal injury group （I）, intimal injury and Le-Mai treatment group （IT）, and vascular endothelial growth factor （20 mg/L） group, Vascular intimal injury was induced by Foley＇s tube in abdominal aorta of rabbits, Le-Mai treatment referred to 25 mg/kg granule fed on rabbits every day. On day 8, one filter membrane of 5 mm diameter was placed on every CAM and 5 μL sample of each group was subtracted onto membrane once a day for three days continuously.③Experimental evaluation： The CAMs were harvested on day 11 and placed on slide after digital camera photograph. The blood vessels around the membrane were counted, And the vascular branch 0,5-com far from the membrane was measured and compared.RESULTS： Eight rabbits and sixty chick embryos were all involved in the result analysis. Compared with normal serum group, the proliferation of blood vessels around CAM was obviously increased in the group T, and the growth presented a convergence trend （P 〈 0.05）; The group Ⅰ showed an extremely significant proliferated vessels compared with normal serum group （P 〈 0.01 ）; There was no significant difference in the total number of blood vessels around CAM between group IT and group Ⅰ.CONCLUSION：①Serum of rabbits fed with Le-Maigranule can stimulate angiogenesis of CAM. ②Serum of rabbits suffering from vascular intimal injury can also stimulate angiogenesis of CAM.