高血压脑出血(HICH)是一种高死亡率的破坏性疾病,发病率,和残疾。积雪草苷(AC)是一种三萜衍生物,已证明对神经元和血管具有保护作用。探讨AC对HICH的作用及可能机制。用20U/mL凝血酶处理人脑微血管内皮细胞(hBMECs)24h,建立体外HICH模型,和浓度为1、2和4μM的AC用于孵育hBMEC。采用细胞计数试剂盒-8、流式细胞术、试管形成试验,血管通透性实验和蛋白质印迹分析。在体内,大鼠注射浓度为150mg/mL的20μL血红蛋白,然后胃内给予1.25、2.5和5mg/kgAC。行为测试,脑含水量测量,苏木精-伊红(HE)染色,末端脱氧核苷酸转移酶脱氧尿苷三磷酸(dUTP)缺口末端标记测定,采用免疫印迹法评价AC对HICH的作用及可能机制。AC(2和4µM)改善了增殖,凋亡,凝血酶诱导的hBMECs中的血管生成和血管通透性(p<0.05)。此外,AC(2.5和5mg/kg)改善了行为评分,脑含水量,病理性病变,HICH大鼠细胞凋亡和血管通透性相关蛋白的表达(p<0.05)。此外,在HICH后,AC在细胞和动物模型中均升高了PI3K/AKT途径的表达(p<0.05)。PI3K/AKT通路抑制剂LY294002的应用,逆转了AC对增殖的改善作用,凋亡,凝血酶诱导的hBMECs中的血管生成和血管通透性(p<0.05)。AC通过增加HICH后PI3K/AKT通路的表达来减轻脑损伤。
Hypertensive intracerebral hemorrhage (HICH) is a destructive disease with high mortality, incidence, and disability. Asiaticoside (AC) is a triterpenoid derivative that has demonstrated to exert a protective effect on neuron and blood vessel. To investigate the function and potential mechanism of AC on HICH. Human brain microvascular endothelial cells (hBMECs) were treated with 20 U/mL thrombin for 24 h to establish the HICH model in vitro, and AC with the concentration of 1, 2 and 4 µM were used to incubate hBMECs. The effect and potential mechanism of AC on HICH were investigated by using cell counting kit-8, flow cytometry, tube forming assays, vascular permeability experiments and western blot assays. In vivo, rats were injected with 20 µL hemoglobin with a concentration of 150 mg/mL, and then intragastrically administrated with 1.25, 2.5 and 5 mg/kg AC. Behavioral tests, brain water content measurement, hematoxylin-eosin (HE) staining, terminal deoxynucleotidyl transferase deoxyuridine triphosphate (dUTP) nick end labeling assays, and western blot were used to assess the effect and potential mechanism of AC on HICH. AC (at 2 and 4 µM) improved the proliferation, apoptosis, angiogenesis and vascular permeability in thrombin-induced hBMECs (p < 0.05). Besides, AC (2.5 and 5 mg/kg) ameliorated behavioral scores, brain water content, pathological lesion, apoptosis and the expression of vascular permeability-related proteins in rats with HICH (p < 0.05). In addition, AC elevated the expression of PI3K/AKT pathway after HICH both in cell and animal models (p < 0.05). Application of LY294002, an inhibitor of PI3K/AKT pathway, reversed the ameliorative effect of AC on the proliferation, apoptosis, angiogenesis and vascular permeability in thrombin-induced hBMECs (p < 0.05). AC reduced brain damage by increasing the expression of the PI3K/AKT pathway after HICH.