疮灵液对模型大鼠血管新生与创面炎症反应的影响

The effects of Chuanglingye decoction on angiogenesis and wound healing

目的：探讨疮灵液对血管新生与创面愈合的影响。方法通过鸡胚尿囊膜模型（CAM）筛选不同剂量的疮灵液对血管新生的疗效；通过血管内皮细胞增殖实验（MTT）、迁移划痕实验探查筛选适合剂量的疮灵液对血管新生的细胞学疗效；采用大鼠背部机械性全层皮肤创面，将生理盐水纱布（对照组）、疮灵液纱布（疮灵液组）覆盖创面，隔日更换；造模时及造模后第3、7、14、28天测定创面面积，造模后第3、7天测炎症反应积分，造模时及造模后第3、7天取创面组织用于检测肿瘤坏死因子（TNF-α）与白细胞介素-6（IL-6）含量。结果 CAM实验显示，疮灵液0.2 ml、0.3 ml组血管新生[分别为（62.6±4.88）个、（62.0±5.70）个]少于对照组（104.0±27.01）个，P＜0.05。选择疮灵液0.2 ml组进行细胞实验，其抑制人脐静脉内皮细胞（HUVEC）增殖（0.66±0.03）%。疮灵液组创面炎症积分治疗后3 d、7 d [分别为（2.91±0.66）分、（1.25±0.45）分]分别低于对照组治疗后3 d、7 d[（4.51±1.01）、（2.25±0.62）分]（P＜0.05或0.01）。创面肉芽TNF-α含量治疗后3 d、7 d[分别为（768±107）ng/L、（380±47）ng/L]，低于对照组治疗后3 d、7 d[分别为（958±140）ng/L、（512±62）ng/L]；IL-6含量治疗后3 d、7 d[分别为（664±133）ng/L、（363±43）ng/L]，低于对照组治疗后3 d、7 d[分别为（2215±314）ng/L、（1562±174）ng/L]（P＜0.05或0.01）。结论疮灵液具有抗血管内皮细胞的迁移与增殖，抑制血管新生作用；且可改善创面的炎症反应，不影响机械性创面的愈合。

Objective To evaluate the effects of Chuanglingye decoction on angiogenesis and wound healing. Methods With a series of dosages of Chuanglingye decoction, their optimal effects of angiogenesis were searched for through the chicken embryo allantois membrane model（CAM）. The vascular endothelial cell proliferation experiment （MTT） and the migration assay were used for the detection of effects. The gauze loading with Chuanglingye decoction of 0.2 ml as the experimental group and with saline of 0.2ml as the control group were applied on the total skin mechanical round wound of 1.5cm diameter and changed every other day. The sizes of area were detected on the day of 0,3,7,14 and 28 as well as the scores of inflammatory response, contains of TNF-αand Il-6 were detected on the day of 3 and 7. Results The CAM experiments showed that the angiogenic effects of 0.2 ml and 0.3 ml dosage of the Chuanglingye group were significantly lower than those of the control group（P〈0.05）. The 0.2 ml dosage of Chuanglingye decoction was chosen for the further experiment. The HUVEC proliferation rate of the experimental group decreased 21%, as compared with the results of control group. The cell migration movement of 12 hours, 24 hours in the experimental group were significantly lower than those in the control group. For theanimal experiments, the area sizes of the wound were＆nbsp;similar in the experimental and control group without any significant differences. The scores of inflammatory response and contains of TNF-α（768±107）ng/L,（380±47）ng/L and Il-6（664±133）ng/L,（363±43）ng/L in the experimental group were significant decreased than those of the control group on the day of 3（958± 140）ng/L,（2215±314）ng/Land 7（512±62）ng/L,（1562±174）ng/L. Conclusion It showed that Chuanglingye decoction had negative effects on vascular endothelial cell migration and proliferation and thus inhibiting angiogenesis. These effects did not infer the process of the wound healing due to its ameliorating the inflammatory response which may be a help to wound healing.