目的 基于高迁移率族蛋白B1（high mobility group box 1，HMGB1）/晚期糖基化终末产物受体（receptor for advanced glycation end products，RAGE）通路探究松萝酸对脑出血（intracerebral hemorrhage，ICH）小鼠炎症损伤的影响。方法 将C57BL/6小鼠随机分为假手术组、模型组、HMGB1抑制剂丙酮酸乙酯（40 mg/kg）组及松萝酸低、高剂量（20、30 mg/kg）组和松萝酸（30 mg/kg）＋rHMGB1（8 μg/kg）组，每组18只。采用颅内注射0.5μL胶原酶构建ICH模型，连续3 d给药后，评价各组小鼠改良神经功能缺损评分（modified neurological severity score，mNSS）和左转率；测定小鼠脑组织含水率；ELISA检测脑组织白细胞介素-6（interleukin-6，IL-6）、IL-1β和IL-18水平；苏木素-伊红（hematoxylin-eosin，HE）染色观察脑组织病理改变；免疫荧光观察脑组织小胶质细胞活化情况；TUNEL染色检测脑组织细胞凋亡；Western blotting检测脑组织HMGB1/RAGE通路相关蛋白表达。结果 与假手术组比较，模型组小鼠纹状体组织损伤严重，mNSS、左侧大脑半球含水率、离子化钙结合适配分子1-（ionized calcium-binding adapter molecule 1，Iba1）阳性细胞数、炎症因子水平、细胞凋亡率及HMGB1、RAGE、磷酸化核因子-κB p65（phosphorylated nuclear factor-κB p65，p-NF-κB p65）/NF-κB p65、肿瘤坏死因子-α（tumor necrosis-α，TNF-α）蛋白表达水平显著升高（P＜0.05），左转率显著降低（P＜0.05）；与模型组比较，松萝酸低、高剂量组和丙酮酸乙酯组纹状体组织损伤程度减轻，mNSS、左侧大脑半球含水率、Iba1阳性细胞数、炎症因子水平、细胞凋亡率及HMGB1/RAGE通路相关蛋白表达水平显著降低（P＜0.05），左转率显著升高（P＜0.05）；rHMGB1显著逆转了松萝酸对ICH小鼠炎症损伤的抑制作用（P＜0.05）。结论 松萝酸可能通过抑制HMGB1/RAGE通路减轻ICH小鼠炎症损伤。

Objective To explore the effect of usnic acid on inflammatory injury in mice with intracerebral hemorrhage (ICH) based on high mobility group box 1 (HMGB1)/receptor for advanced glycation end products (RAGE) pathway. Methods C57BL/6 mice were randomly divided into sham group, model group, HMGB1 inhibitor ethyl pyruvate (40 mg/kg) group, usnic acid low-, high-dose (20, 30 mg/kg) groups and usnic acid (30 mg/kg) + rHMGB1 (8 μg/kg) group, with 18 mice in each group. The ICH model was constructed by intracranial injection of 0.5 μL collagenase. After continuous administration for 3 d, the modified neurological severity score (mNSS) and left turn rate of mice in each group were evaluated. The water rate of brain tissue in mice was determined. ELISA was used to measure the levels of interleukin-6 (IL-6), IL-1β and IL-18 in brain tissue. Hematoxylin-eosin (HE) staining was used to observe pathological changes in brain tissue. Immunofluorescence was used to observe microglia activation in brain tissue. TUNEL staining was used to detect cell apoptosis of brain tissue. Western blotting was used to detect the expressions of HMGB1/RAGE pathway related proteins in brain tissue. Results Compared with sham group, the striatum tissue of mice in model group was severely damaged, mNSS, left cerebral hemisphere water rate, ionized calcium-binding adapter molecule 1 (Iba1) positive cells number, inflammatory factors levels, cell apoptosis rate, HMGB1, RAGE, phosphorylated nuclear factor-κB p65 (p-NF-κB p65)/NF-κB p65 and TNF-α protein expression levels were significantly increased (P < 0.05), and the left turn rate was significantly decreased (P < 0.05). Compared with model group, the damage degree of striatum tissue in usnic acid low-, high-dose group and ethyl pyruvate group was alleviated (P < 0.05), mNSS, left cerebral hemisphere water rate, Iba1 positive cells number, inflammatory factors levels, cell apoptosis rate and HMGB1/RAGE pathway related proteins expression levels were significantly decreased (P < 0.05), and the left turn rate was significantly increased (P < 0.05). rHMGB1 significantly reversed the inhibitory effect of usnic acid on inflammatory injury in ICH mice (P < 0.05). Conclusion Usnic acid may alleviate inflammatory injury in ICH mice by inhibiting HMGB1/RAGE pathway.

R285.5

河南省科学技术厅科技攻关项目（252102310266）