目的 基于接触系统探讨僵蚕提取液激活凝血因子XII（coagulation factor XII，FXII）对缺血性脑卒中大鼠血管新生的干预作用。方法 检测僵蚕提取液干预血浆凝血四项，体外检测僵蚕提取液对FXII及其下游底物激肽释放酶（kallikrein-kinin，KK）的激活作用。通过大鼠脑缺血再灌注损伤模型，检测大鼠的脑形态学及FXII、KK、血管内皮生长因子（vascular endothelial cell growth factors，VEGF）、CD31、Brdu⁺/vWF⁺蛋白表达；通过氧糖剥夺模型，检测大鼠脑微血管内皮细胞的形态学改变、细胞增殖情况及VEGF的表达。采用LC-MS鉴定僵蚕提取液药效物质。结果 在体外实验中，僵蚕提取液延长了人血浆部分凝血酶原时间（activated partial thromboplastin time，APTT）、凝血酶原时间（prothrombin time，PT）、凝血酶时间（thrombin time，TT）（P＜0.05），激活FXII并导致下游底物KK的生成，呈剂量相关性（P＜0.05）；在大鼠脑缺血再灌注模型中，僵蚕提取液改善了大鼠神经元损伤，激活了FXII，导致KK生成和VEGF、CD31、Brdu⁺/vWF⁺的表达增加（P＜0.05、0.01）；在细胞氧糖剥夺实验中，僵蚕提取液干预提高了大鼠脑微血管内皮细胞的增殖和VEGF的表达（P＜0.05）。LCMS鉴定僵蚕提取液含代谢物共计809个。结论 僵蚕提取液可改善缺血再灌注大鼠神经功能损伤，其机制可能与抗凝和促脑微血管内皮细胞增殖有关。

Objective To explore the interventional effect of Jiangcan (Bombyx Batryticatus) extracts as an activator of coagulation factor Ⅻ (FⅫ) on angiogenesis of rats with ischemic stroke by contact system.Methods The effects of Bombyx Batryticatus extract on plasma coagulation were detected, and the activation of FXII and its downstream substrate kallikrein kinase (KK) by Bombyx Batryticatus extract was detected in vitro. The brain morphology and expressions of FXII, KK, vascular endothelial growth factors (VEGF), CD31, Brdu⁺/vWF⁺ were detected by rats model of cerebral ischemia reperfusion injury; The morphological changes, cell proliferation and VEGF expression of brain microvascular endothelial cells were detected by oxygen glucose deprivation model. The pharmacodynamic substances of Bombyx Batryticatus extract were identified by LC-MS.Results In vitro, Bombyx Batryticatus extract prolonged the activated partial thromboplastin time (APTT), prothrombin time (PT), thrombin time (TT) (P<0.05), activated FⅫ and promoted the production of downstream substrate KK, with dose-dependent (P<0.05). In the model of cerebral ischemia reperfusion in rats, Bombyx Batryticatus extract improved the neuronal damage of rats, activated FXII and increased the production of KK and the expressions of VEGF, CD31, Brdu⁺/vWF⁺ (P<0.05, 0.01); In the cell oxygen and glucose deprivation experiment, Bombyx Batryticatus extract increased the proliferation of brain microvascular endothelial cells and expression of VEGF in rats (P<0.05). A total of 809 metabolites in Bombyx Batryticatus extract were identified by LC-MS.Conclusion Bombyx Batryticatus extract can ameliorate the injury of nerve function in rats with ischemia-reperfusion, and its mechanism may be related to anticoagulation and promoting the proliferation of brain microvascular endothelial cells.

R285.5

湖南省自然科学基金资助项目(2020JJ5424);湖南省教育厅青年基金资助项目(21B30386);湖南中医药大学中西医结合“双一流”学科开放基金资助项目(2020ZXYJH08)