目的 基于腺苷酸活化蛋白激酶（AMPK）-过氧化物酶体增殖物激活受体 γ 共激活因子 1α（PGC1α）信号 轴探讨安宫牛黄丸减轻氧化应激改善脑出血小鼠神经功能损伤的作用机制。方法 采用脑立体定位法在小鼠左 侧尾状壳核注射 0.5 µL Ⅶ-S 型胶原酶（0.15 U·µL- ¹）复制脑出血小鼠模型。将 93 只 C57BL/6 小鼠随机分为假 手术组、模型组与安宫牛黄丸低、中、高剂量组（20、40、80 mg·kg-1 ），以及安宫牛黄丸（40 mg·kg-1 ）+抑制剂 组（Dorsomorphin 10 mg·kg-1 ，造模前 1 h 腹腔注射），每组 5～6 只。安宫牛黄丸给药组按上述不同剂量连续灌 胃干预 3 d，每日 1 次。采用改良神经功能缺损评分（NDS）及转角实验评估小鼠的神经功能，并测定脑含水量； FJB 染色法分析神经元退变情况；比色法检测脑组织氧化应激指标丙二醛（MDA）、超氧化物歧化酶（SOD）、谷 胱甘肽（GSH）、氧化型谷胱甘肽（GSSG）的水平；Western Blot 法检测脑组织磷酸化腺苷酸活化蛋白激酶（pAMPK）、PGC1α、核因子 E2 相关因子 2（Nrf2）蛋白表达水平。结果 与假手术组比较，模型组小鼠脑出血 24、 72 h 的 p-AMPK 蛋白表达显著上调（P＜0.05，P＜0.01），脑出血 72 h 的 PGC1α 蛋白表达显著上调（P＜0.01）， Nrf2 蛋白表达有上调趋势（P＞0.05）；NDS 评分及自主运动左转率均显著升高（P＜0.01）；同侧（左侧）大脑半球 的脑含水量显著升高（P＜0.01）；脑组织中 MDA、GSSG 水平显著升高（P＜0.01），SOD、GSH 水平显著降低 （P＜0.01）。与模型组比较，安宫牛黄丸中、高剂量组小鼠的 NDS 评分显著降低（P＜0.05，P＜0.01），但安宫 牛黄丸各剂量组小鼠的自主运动左转率无明显变化（P＞0.05）；安宫牛黄丸中剂量组小鼠同侧大脑半球的脑含 水量、FJB 阳性细胞数量显著降低（P＜0.01），脑组织中 MDA、GSSG 水平显著降低（P＜0.05，P＜0.01）， SOD、GSH 水平明显升高（P＜0.05），p-AMPK、PGC1α、Nrf2 蛋白表达均显著上调（P＜0.05，P＜0.01）。与安 宫牛黄丸中剂量组比较，安宫牛黄丸+抑制剂组小鼠脑组织中 p-AMPK、PGC1α、Nrf2 蛋白表达水平均显著降 低（P＜0.05，P＜0.01）。结论 安宫牛黄丸能减轻脑出血小鼠的局部脑水肿，减少神经元变性，改善神经功能 损伤，可能与调控 AMPK-PGC1α 信号轴减轻脑组织的氧化应激损伤有关。

Objective To investigate the mechanism by which Angong Niuhuang Pills alleviates neurological impairment in mice with intracerebral hemorrhage （ICH） by reducing oxidative stress， based on the adenosine monophosphate-activated protein kinase （AMPK）-peroxisome proliferator-activated receptor gamma coactivator 1- alpha （PGC1α） signaling axis. Methods An ICH mouse model was established by stereotactically injecting 0.5 µL of type VII-S collagenase （0.15 U·µL- ¹） into the left caudate putamen of mice. Ninety-three C57BL/6 mice were randomly divided into the following groups： sham-operated group， model group， low- ， medium- ， and high-dose Angong Niuhuang Pills groups （20，40，80 mg·kg⁻¹，respectively），and an Angong Niuhuang Pills （40 mg·kg⁻¹） + inhibitor group （Dorsomorphin 10 mg·kg⁻¹，intraperitoneally injected 1-hour before modeling），with 5-6 mice per group. Mice in the Angong Niuhuang Pills administration groups received the respective doses via oral gavage once daily for 3 consecutive days. Neurological function was assessed using the modified Neurological Deficit Score （NDS） and corner test， and brain water content was measured. Neuronal degeneration was analyzed by Fluoro-Jade B （FJB） staining. Oxidative stress markers in brain tissue， including malondialdehyde （MDA）， superoxide dismutase （SOD）， glutathione （GSH），and oxidized glutathione （GSSG），were detected by colorimetric assays. Protein expression levels of phosphorylated AMPK （p-AMPK），PGC1α，and nuclear factor erythroid 2-related factor 2（Nrf2） in brain tissue were detected by Western Blot. Results Compared with the sham-operated group，the expression of p-AMPK protein in the model group was significantly upregulated at 24 hours and 72 hours post-ICH （P＜0.05， P＜0.01）， PGC1α protein expression was significantly upregulated at 72 hours post-ICH （P＜0.01），and Nrf2 protein expression showed an upward trend （P＞0.05）. The NDS scores and the rate of autonomous left turns were significantly increased （P＜ 0.01）. The brain water content in the ipsilateral （left） cerebral hemisphere was significantly elevated （P＜0.01）. Levels of MDA and GSSG in brain tissue were significantly increased （P＜0.01）， while levels of SOD and GSH were significantly decreased （P＜0.01）. Compared with the model group， mice in the medium- and high-dose Angong Niuhuang Pills groups showed significantly reduced NDS scores （P＜0.05，P＜0.01），although the autonomous left turn rates did not change significantly across all Angong Niuhuang Pills dose groups （P＞0.05）. In the medium-dose Angong Niuhuang Pills group，the brain water content in the ipsilateral hemisphere and the number of FJB-positive cells were significantly reduced （P＜0.01）；levels of MDA and GSSG in brain tissue were significantly decreased （P＜ 0.05， P＜0.01）， while SOD and GSH levels were significantly increased （P＜0.05）. Furthermore， the protein expression of p-AMPK， PGC1α， and Nrf2 was significantly upregulated （P＜0.05， P＜0.01）. Compared with the medium-dose Angong Niuhuang Pills group，the Angong Niuhuang Pills + inhibitor group showed significantly reduced protein expression levels of p-AMPK，PGC1α，and Nrf2 in brain tissue （P＜0.05，P＜0.01）. Conclusion Angong Niuhuang Pills can alleviate local cerebral edema， reduce neuronal degeneration， and improve neurological impairment in mice with ICH. This effect may be associated with the modulation of the AMPK-PGC1α signaling axis to attenuate oxidative stress damage in brain tissue.

R285.5

“十四五”国家重点研发计划“中医药现代化研究”重点专项（2022YFC3501103）；河南省中医药科学研究专项重点课题（2024ZY1007）； 河南省高等学校重点科研项目（22A320037）；河南省中医药科学研究专项课题（2022ZY1009）。