目的 探究生、熟大黄在脑中动脉栓塞再灌注（middle cerebral artery occlusion and reperfusion，MCAO/R）模型大鼠体内药物代谢的差异性。方法 线栓法构建MCAO/R大鼠模型，将造模成功的大鼠随机分为模型组、生大黄（2.5 g/kg）组和熟大黄（2.5 g/kg）组，每组6只，大鼠连续给药7 d。末次给药前禁食12 h，给药1 h后，采集血液及脑组织。采用超高效液相色谱-四极杆-静电场轨道阱-线性离子阱质谱（ultra-high performance liquid chromatography-quadrupole electrostatic field orbitrap linear ion trap mass spectrometry，UPLC-Orbitrap Fusion Lumos Tribrid-MS）表征生、熟大黄在MCAO/R大鼠体内（血浆、脑组织）中的原型及其代谢产物，通过多元统计分析筛选差异性成分，并结合解析电离喷雾电离-质谱成像技术（desorption electrospray ionization mass spectrometry imaging，DESI-MSI）对脑组织中生、熟大黄差异成分的原位空间分布进行可视化分析。结果 在MCAO/R大鼠体内中共鉴定109个原型成分和代谢产物，根据变量权重值（variable important in projection，VIP）＞1、P＜0.05，从血浆中和脑组织中分别筛选出生、熟大黄有23、8个差异性成分，熟大黄组血浆中蒽醌苷类减少而脑组织中苷元及代谢物（如没食子酸、M31）增加，且主要分布在脑组织中的海马区、中脑及皮质区；共有差异成分没食子酸和代谢产物M31在熟大黄组显著上升，在纹状体呈现特异性富集。结论 对生、熟大黄在MCAO/R大鼠体内的药物代谢差异性成分进行表征及空间定位，发现熟大黄能够提高脑内活性苷元和代谢物暴露水平，增强其对脑组织的保护作用，为“生熟异治”的炮制理论提供科学依据。

Objective To explore the differences in the in vivo drug metabolism of raw and processed Dahuang (Rhei Radix et Rhizoma) in a rat model of middle cerebral artery occlusion and reperfusion (MCAO/R). Methods The MCAO/R rat model was established by the suture method. The successfully modeled rats were randomly divided into model group, raw Rhei Radix et Rhizoma (2.50 g/kg) group and processed Rhei Radix et Rhizoma group, with six rats in each group. Rats were administered continuously for 7 d. 12 h before the last administration, the rats were fasted, and 1 h after administration, blood and brain tissues were collected. Ultra-high performance liquid chromatography-quadrupole electrostatic field orbitrap linear ion trap mass spectrometry (UPLC-Orbitrap Fusion Lumos Tribrid-MS) was used to characterize the prototypes and metabolites of raw and processed Rhei Radix et Rhizoma in plasma and brain tissues of MCAO/R rats. Multivariate statistical analysis was used to screen the differential components, and desorption electrospray ionization mass spectrometry imaging (DESI-MSI) technique was combined to visualize the in situ spatial distribution of the differential components of raw and processed Rhei Radix et Rhizoma in brain tissue. Results A total of 109 prototype components and metabolites were identified in MCAO/R rats. According to variable important in projection (VIP) > 1 and P < 0.05, 23 and eight differential components of raw and processed Rhei Radix et Rhizoma were screened from the plasma and brain tissues, respectively. In the plasma of processed Rhei Radix et Rhizoma group, anthraquinone glycosides were decreased, while in the brain tissue, aglycones and metabolites (such as gallic acid, M31) were increased, and were mainly distributed in hippocampus, midbrain and cortex of brain tissue. The common differential components gallic acid and metabolite M31 were significantly increased in the processed Rhei Radix et Rhizoma group and were specifically enriched in the striatum. Conclusion This study characterized and spatially located the differential components of raw and processed Rhei Radix et Rhizoma in MCAO/R rats and found that processed Rhei Radix et Rhizoma increased the exposure levels of active aglycones and metabolites in the brain, enhancing its protective effect on brain tissue, providing a scientific basis for the “different treatment for raw and processed” processing theory.

R285.61

河南省中医药科研专项课题（2024ZY1033）