摘要
目的:探讨清热开窍方对缺血性脑卒中(ischemic stroke,IS)大鼠的神经保护作用,并基于核因子E2相关因子2(nuclear factor erythroid 2-related factor 2,Nrf2)/血红素加氧酶-1(heme oxygenase-1,HO-1)信号通路探讨相关作用机制。方法:将75只雄性SD大鼠随机分为假手术组(Sham组)、模型组(MCAO组)、清热开窍方组(QRKQF组,9.14 g·kg^(-1))、清热开窍方+Nrf2抑制剂组(QRKQF+ML385组)、Nrf2抑制剂组(ML385组,30 mg·kg^(-1)),每组15只。除Sham组外,其余大鼠采用线栓法建立大脑中动脉栓塞(middle cerebral artery occlusion,MCAO)模型。采用Garcia JH法评价大鼠的神经功能;TTC染色法检测脑缺血体积;干湿重法检测脑含水量;Western blot法检测皮质Nrf2、HO-1、核苷酸结合寡聚化结构域样受体蛋白3(nucleotide-binding oligomerization domain-like receptor protein 3,NLRP3)、裂解的半胱天冬酶-1(cleaved cysteinyl aspartate specific proteinase-1,c-Caspase-1)、消皮素D-N(gasdermin D-N,GSDMD-N)蛋白表达水平;试剂盒检测皮质丙二醛(malondialdehyde,MDA)、谷胱甘肽(glutathione,GSH)、超氧化物歧化酶(superoxide dismutase,SOD)、过氧化氢酶(catalase,CAT)水平。结果:与Sham组比较,MCAO组大鼠Garcia JH评分降低、脑缺血体积与脑含水量增加(P<0.01);与MCAO组比较,QRKQF组大鼠Garcia JH评分升高、脑缺血体积与脑含水量减少(P<0.01);与ML385组比较,QRKQF+ML385组大鼠Garcia JH评分升高、脑缺血体积与脑含水量减少(P<0.05)。与MCAO组比较,QRKQF组大鼠皮质Nrf2、HO-1蛋白表达水平升高,NLRP3、c-Caspase-1、GSDMD-N蛋白表达水平降低(P<0.01);与ML385组比较,QRKQF+ML385组大鼠皮质Nrf2、HO-1蛋白表达水平升高,NLRP3、c-Caspase-1、GSDMD-N蛋白表达水平降低(P<0.01)。与Sham组比较,MCAO组大鼠皮质MDA水平升高,GSH、SOD、CAT水平降低(P<0.01);与MCAO组比较,QRKQF组大鼠皮质MDA水平降低,GSH、SOD、CAT水平升高(P<0.01);与ML385组比较,QRKQF+ML385组大鼠皮质MDA水平降低,GSH、SOD、CAT水平升高(P<0.01)。结论:清热开窍方通过抑制IS大鼠细胞焦亡发挥神经保护作用,这可能与激活Nrf2/HO-1信号通路有关。
Objective:To investigate the neuroprotective effect of Qingre Kaiqiao Formula on ischemic stroke(IS)rats,and to explore the mechanism based on nuclear factor erythroid 2-related factor 2(Nrf2)/heme oxygenase-1(HO-1)signaling pathway.Methods:Seventy-five male SD rats were randomly divided into sham operation group(Sham group),model group(MCAO group),Qingre Kaiqiao Formula group(QRKQF group,9.14 g·kg^(-1)),Qingre Kaiqiao Formula Nrf2 inhibitor group(QRKQF ML385 group)plus Nrf2 inhibitor group(ML385 group,30 mg·kg^(-1)),with 15 rats in each group.Except for the Sham group,the other rats used the wire thrombosis method to establish a middle cerebral artery occlusion(MCAO)model.Garcia JH method was used to evaluate the neurological function of rats.TTC staining was used to detect cerebral ischemic volume.Brain water content was detected by dry and wet gravimetric method;Cortical Nrf2,HO-1,nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3),and cleaved cysteinyl aspartate specific were detected by Western blot proteinase-1,c-caspase-1),gasdermin D-N(GSDMD-N)protein expression levels;The kit detected the levels of malondialdehyde(MDA),glutathione(GSH),superoxide dismutase(SOD),and catalase(CAT).Results:Compared with the Sham group,the Garcia JH score of the rats in the MCAO group decreased,and the cerebral ischemic volume and cerebral water content increased(P<0.01).Compared with the MCAO group,the Garcia JH score of the rats in the QRKQF group was increased,and the cerebral ischemic volume and cerebral water content were decreased(P<0.01).Compared with the ML385 group,the Garcia JH score,cerebral ischemia volume and cerebral water content of the rats in the QRKQF ML385 group were increased(P<0.05).Compared with the MCAO group,the expression levels of cortical Nrf2 and HO-1 proteins in the QRKQF group were increased,and the expression levels of NLRP3,c-Caspase-1 and GSDMD-N proteins decreased(P<0.01).Compared with the ML385 group,the expression levels of cortical Nrf2 and HO-1 proteins in the QRKQF ML385 group were increased,and the expression levels of NLRP3,c-caspase-1 and GSDMD-N proteins were decreased(P<0.01).Compared with the Sham group,the levels of cortical MDA in the MCAO group were increased,and the levels of GSH,SOD and CAT decreased(P<0.01).Compared with the MCAO group,the levels of cortical MDA in the QRKQF group decreased,and the levels of GSH,SOD and CAT increased(P<0.01).Compared with the ML385 group,the levels of cortical MDA in the QRKQF ML385 group were decreased,and the levels of GSH,SOD and CAT were increased(P<0.01).Conclusion:Qingre Kaiqiao Formula exerts a neuroprotective effect by inhibiting pyroptosis in IS rats,which may be related to the activation of Nrf2/HO-1 signaling pathway.
作者
刘嘉
刘云婷
鱼童
张紫涵
刘雪梅
LIU Jia;LIU Yunting;YU Tong;ZHANG Zihan;LIU Xuemei(Dongfang Hospital,Beijing University of Chinese Medicine,Beijing China 100078)
出处
《中医学报》
2025年第9期1960-1967,共8页
Acta Chinese Medicine
基金
国家自然科学基金项目(81973788,82374405)。
