紫草素调节HIF-1α/NLRP3信号通路对蛛网膜下腔出血大鼠神经功能损伤的影响  

Impact of shikonin on neural function damage in rats with subarachnoid hemorrhage by regulating the HIF-1α/NLRP3 signaling pathway

作　　者：饶重贤 胡姗姗 谭伟[2] 王军民[3] 金胜昔[4] 周游[4] RAO Chongxian;HU Shanshan;TAN Wei(Brain Center,Geriatric Hospital Affiliated to Wuhan University of Science and Technology,Hubei,Wuhan 430065,China;不详)

机构地区：[1]武汉科技大学附属老年病医院脑科中心,武汉市430065 [2]武汉科技大学附属老年病医院全科医学科,武汉市430065 [3]武汉大学人民医院神经外科 [4]武汉科技大学附属天佑医院神经外科

出　　处：《河北医药》2024年第4期496-500,505,共6页Hebei Medical Journal

基　　金：湖北省卫生健康委中医药科研立项项目(编号:ZY2021M074)。

摘　　要：目的探究紫草素调节缺氧诱导因子-1α(HIF-1α)/NOD样受体热蛋白结构域相关蛋白3(NLRP3)信号通路对蛛网膜下腔出血(SAH)大鼠神经功能损伤的影响。方法大鼠随机分为假手术组、模型组、YC-1(HIF-1α抑制剂,5 mg/kg)组、紫草素低剂量组(4 mg/kg)、紫草素高剂量组(25 mg/kg)、紫草素高剂量(25 mg/kg)+AG1(HIF-1α激活剂,10 mg/kg)组,每组15只。采用颈内动脉刺破法制备SAH模型。采用Zea-Longa评分法评估6组大鼠神经功能;ELISA法检测血清肿瘤坏死因子α(TNF-α)、白介素-6(IL-6)和IL-1β水平;HE染色观察大鼠海马组织形态学变化,TUNEL染色法检测海马神经元凋亡率;伊文思蓝染色检测血脑屏障通透性;商品化试剂盒检测大鼠脑组织超氧化物歧化酶(SOD)、过氧化氢酶(MDA)、丙二醛(CAT)水平,Western blot检测大鼠脑组织Bax、Bcl-2、HIF-1α、NLRP3蛋白表达。结果假手术组大鼠海马神经元形态结构正常;与假手术组相比,模型组大鼠海马神经元有大量水肿、结构模糊,有细胞核溶解,变型固缩,部分细胞核消失,大鼠神经功能评分、血清TNF-α、IL-6和IL-1β水平、海马神经元凋亡率、伊文思蓝渗出量、脑组织MDA水平、Bax、HIF-1α、NLRP3水平显著增加,脑组织SOD、CAT水平、Bcl-2蛋白水平显著降低(P<0.05);与模型组比较,紫草素低剂量组、紫草素高剂量组和YC-1组鼠海马神经元病理损伤显著改善,大鼠神经功能评分、血清TNF-α、IL-6和IL-1β水平、海马神经元凋亡率、伊文思蓝渗出量、脑组织MDA水平、Bax、HIF-1α、NLRP3水平显著降低,SOD、CAT水平、Bcl-2蛋白水平显著升高(P<0.05);与紫草素高剂量组比较,紫草素高剂量+AG1组大鼠海马神经元病理损伤显著加重,大鼠神经功能评分、血清TNF-α、IL-6和IL-1β水平、海马神经元凋亡率、伊文思蓝渗出量、脑组织MDA水平、Bax、HIF-1α、NLRP3水平显著增加,SOD、CAT水平、Bcl-2蛋白水平显著降低(P<0.05)。Objective To investigate the impact of shikonin on neural function damage in rats with subarachnoid hemorrhage(SAH)by regulating the hypoxia inducible factor-1α(HIF-1α)/NOD-like receptor pyrin domain containing 3(NLRP3)signaling pathway.Methods Rats were randomly grouped into the sham surgery group,model group,YC-1(HIF-1αinhibitor,5mg/kg)group,low-dose shikonin group(4mg/kg),high-dose shikonin group(25mg/kg),and high-dose shikonin(25mg/kg)+AG1(HIF-1αactivator,10mg/kg)group,with 15 rats peRgroup.The internal carotid artery puncture method was applied to prepare a SAH model.The Zea Longa scoring method was applied to evaluate the neurological function of rats in each group.The enzyme-linked immunosorbent assay(ELISA)method was applied to detect serum levels of tumoRnecrosis factor alpha(TNF-α),interleukin-6(IL-6),and IL-1β.Morphological changes of hippocampal tissues,apoptosis rate of hippocampal neurons and blood-brain barrieRpermeability were detected by hematoxylin and eosin(H&E)staining,terminal deoxynucleotidyl transferase dUTP nick end labeling(TUNEL)staining and Evans blue staining,respectively.Commercial reagent kits were used to detect the levels of superoxide dismutase(SOD),malondialdehyde(MDA)and catalase(CAT)in rat brain tissue.Western blot was applied to detect the protein expressions of Bax,Bcl-2,HIF-1α,and NLRP3 in rat brain tissue.Results The morphology and structure of hippocampal neurons in the sham operation group were intact and clear.Compared with those of the sham surgery group,rats in the model group presented a large amount of edema,blurred structure,nucleaRdissolution,deformation,pyknosis and the disappearance of some nuclei in hippocampal neurons,significantly higher Zea Longa score,serum TNF-α,IL-6 and IL-1βlevels,apoptosis rate of hippocampal neurons,Evans blue exudation amount,MDA level and protein expressions of Bax,HIF-1α,and NLRP3,but significantly lower levels of SOD and CAT and protein expression of Bcl-2(P<0.05).Compared with those of the model group,rats in the low-dose

关 键 词：紫草素 HIF-1α/NLRP3信号通路 蛛网膜下腔出血 神经功能损伤 

分 类 号：R743.35[医药卫生—神经病学与精神病学]