Hou Yongchun, Yan Zi, Wan Haitong, Yang Jiehong, Ding Zhishan, He Yu
Basic Medical School, Zhejiang Chinese Medical University, Hangzhou 310053, China.
Key Laboratory of TCM Encephalopathy of Zhejiang Province, Hangzhou 310053, China.
Brain Sci. 2024 Jul 31;14(8):781. doi: 10.3390/brainsci14080781.
Cerebral ischemia-reperfusion injury (IRI), occurring after blood supply restoration, contributes significantly to stroke-related deaths. This study explored the combined impact and mechanisms of astragaloside IV (AS-IV), hydroxysafflor yellow A (HSYA), and their combination in mitigating IRI. Male Sprague-Dawley (SD) rats were randomized to the Sham, MCAO, MCAO+AS-IV, MCAO+HSYA, and MCAO+AS-IV+HSYA groups. Neurological deficits and cerebral infarction were examined after restoring the blood supply to the brain. Pathomorphological changes in the cerebral cortex were observed via HE staining. IL-1β and IL-18 were quantified using ELISA. The expression of NF-κB and GSDMD in the ischemic cerebrum was analyzed using immunohistochemistry. The expression levels of NLRP3, ASC, IL-1β, Caspase-1, and GSDMD in the ischemic cerebrum were evaluated using Western blot. The MCAO+AS-IV, MCAO+HSYA, and MCAO+AS-IV+HSYA groups exhibited notably better neurological function and cerebral infarction compared with the MCAO group. The combined treatment demonstrated superior brain tissue injury alleviation. Reductions in NF-κB, GSDMD positive cells, and NLRP3/ASC/IL-1β/Caspase-1/GSDMD protein expression in the ischemic brain were significantly more pronounced with the combined therapy, indicating a synergistic effect in countering cerebral IRI via the NF-κB/NLRP3/Caspase-1/GSDMD pathway inhibition of cell pyroptosis-induced injury.
脑缺血再灌注损伤(IRI)发生于血供恢复后,是导致卒中相关死亡的重要原因。本研究探讨了黄芪甲苷(AS-IV)、羟基红花黄色素A(HSYA)及其组合对减轻IRI的联合影响及机制。将雄性Sprague-Dawley(SD)大鼠随机分为假手术组、大脑中动脉闭塞(MCAO)组、MCAO+AS-IV组、MCAO+HSYA组和MCAO+AS-IV+HSYA组。在恢复脑血供后检测神经功能缺损和脑梗死情况。通过苏木精-伊红(HE)染色观察大脑皮质的病理形态学变化。采用酶联免疫吸附测定(ELISA)法对白细胞介素-1β(IL-1β)和白细胞介素-18(IL-18)进行定量分析。采用免疫组织化学法分析缺血脑组织中核因子-κB(NF-κB)和Gasdermin D(GSDMD)的表达。采用蛋白质免疫印迹法评估缺血脑组织中NLRP3、凋亡相关斑点样蛋白(ASC)、IL-1β、半胱天冬酶-1(Caspase-1)和GSDMD的表达水平。与MCAO组相比,MCAO+AS-IV组、MCAO+HSYA组和MCAO+AS-IV+HSYA组的神经功能和脑梗死情况明显更好。联合治疗在减轻脑组织损伤方面表现更优。联合治疗使缺血脑组织中NF-κB、GSDMD阳性细胞以及NLRP3/ASC/IL-1β/Caspase-1/GSDMD蛋白表达的降低更为显著,表明其通过抑制NF-κB/NLRP3/Caspase-1/GSDMD途径减轻细胞焦亡诱导的损伤,从而对脑IRI具有协同作用。