目的 探讨疏血通注射液是否通过抑制泛凋亡减轻大鼠脑缺血再灌注损伤。方法 雄性SD大鼠随机分为假手术组、模型组及疏血通注射液高、中、低剂量（1.08、0.54、0.27 mL/kg）组和依达拉奉（1.35 mL/kg）组，通过短暂性大脑中动脉闭塞制备脑缺血再灌注损伤大鼠模型，术后24 h进行指标评定及取材。神经功能评分检测神经功能；2,3,5-三苯基氯化四氮唑（2,3,5-triphenyltetrazolium chloride solution，TTC）法检测脑梗死面积；苏木素-伊红（hematoxylin eosin，HE）染色和尼氏染色观察脑组织形态结构；透射电镜观察神经元、小胶质细胞和星形胶质细胞的超微结构；TUNEL染色检测细胞凋亡率；碘化丙啶（propidium iodide，PI）染色检测细胞坏死情况；生化试剂盒检测血清和皮层丙二醛（malondialdehyde，MDA）含量和谷胱甘肽过氧化物酶（glutathione peroxidase，GSH-Px）活性；ELISA检测血清和皮层白细胞介素-1β（interleukin-1β，IL-1β）、IL-6和大鼠肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）含量；qRT-PCR和Western blotting检测脑组织核苷酸结合寡聚化结构域样受体蛋白3（NOD-like receptor protein 3，NLRP3）、半胱氨酸天冬氨酸蛋白酶-1（cystein-asparate protease-1，CASP1）、消皮素D（gasdermin D，GSDMD）、CASP8、B细胞淋巴瘤-2（B-cell lymphoma-2，Bcl-2）、Bcl-2相关X蛋白（Bcl-2 associated X protein，Bax）、受体相互作用丝氨酸/苏氨酸激酶1（receptor interacting serine/threonine kinase 1，RIPK1）、RIPK3、混合谱系激酶结构域样蛋白（mixed lineage kinase domain-like protein，MLKL）、Toll样受体2（Toll-like receptor 2，TLR2）、TLR4的mRNA和蛋白表达。结果 与模型组比较，疏血通注射液高剂量组大鼠神经功能评分和脑梗死率显著降低（P＜0.01），脑组织病理学改变减轻；神经元、小胶质细胞和星形胶质细胞体积减少，细胞肿胀情况减轻，细胞膜破裂减少，细胞凋亡率和坏死率显著降低（P＜0.01）；大鼠血清和皮层中MDA、IL-1β、IL-6和TNF-α含量显著降低（P＜0.01），GSH-Px活性显著升高（P＜0.01）；皮层中Bcl-2的mRNA和蛋白表达水平显著升高（P＜0.05、0.01），NLRP3、CASP1、GSDMD、CASP8、Bax、RIPK1、RIPK3、MLKL、TLR2、TLR4的mRNA和蛋白表达水平显著降低（P＜0.05、0.01）。结论 疏血通注射液可能通过抑制泛凋亡，减轻大鼠脑缺血再灌注损伤。

Objective To investigate whether Shuxuetong Injection (疏血通注射液) can alleviate cerebral ischemia-reperfusion injury in rats by inhibiting PANoptosis. Methods Male SD rats were randomly divided into sham group, model group, Shuxuetong Injection high-, medium-, low-dose (1.08, 0.54, 0.27 mL/kg) groups and edaravone (1.35 mL/kg) group. A rat model of cerebral ischemia-reperfusion injury was prepared by transient middle cerebral artery occlusion, and indicators were evaluated and samples were taken 24 h after surgery. Nerve function score was used to detect nerve function. The cerebral infarction area was measured by 2,3,5-triphenyltetrazolium chloride solution (TTC). The morphological structure of brain tissue was observed by hematoxylin-eosin (HE) staining and Nissl staining. The ultrastructure of neurons, microglia and astrocytes were observed by transmission electron microscopy. TUNEL staining was used to detect apoptosis rate. Propidium iodide (PI) staining was used to detect cell necrosis. Content of malondialdehyde (MDA) and glutathione peroxidase (GSH-Px) activity in serum and cortex were detected by biochemical reagent kits. Content s of interleukin-1β (IL-1β), IL-6 and tumor necrosis factor-α (TNF-α) in serum and cortex were detected by ELISA. The mRNA and protein expressions of NOD-like receptor protein 3 (NLRP3), cystein-asparate protease-1 (CASP1), gasdermin D (GSDMD), CASP8, B-cell lymphoma-2 (Bcl-2), Bcl-2 associated X protein (Bax), receptor interacting serine/threonine kinase 1 (RIPK1), RIPK3, mixed lineage kinase domain-like protein (MLKL), Toll-like receptor 2 (TLR2) and TLR4 in brain tissue were detected by qRT-PCR and Western blotting. Results Compared with model group, the neurological function score and cerebral infarction rate of rats in Shuxuetong Injection high-dose group were significantly decreased (P < 0.01), and pathological changes in brain tissue was reduced. The volume of neurons, microglia and astrocytes were decreased, the swelling of cells was decreased, the rupture of cell membranes was decreased, and the rates of apoptosis and necrosis were significantly decreased (P < 0.01). The levels of MDA, IL-1β, IL-6 and TNF-α in serum and cortex of rats were significantly reduced (P < 0.01), while the activity of GSH-Px was significantly increased (P < 0.01). The mRNA and protein expression levels of Bcl-2 in cortex were significantly increased (P < 0.05, 0.01), while the mRNA and protein expression levels of NLRP3, CASP1, GSDMD, CASP8, Bax, RIPK1, RIPK3, MLKL, TLR2, TLR4 were significantly reduced (P < 0.05, 0.01). Conclusion Shuxuetong Injection may alleviate cerebral ischemia-reperfusion injury in rats by inhibiting PANoptosis.

R285.5

国家自然科学基金青年科学基金项目（82003960）