摘要
目的:分析金丝桃苷(Hyp)调节硫氧还蛋白结合蛋白(TXNIP)/核苷酸结合寡聚结构域样受体蛋白3(NLRP3)信号通路对细菌性脑膜炎(BM)大鼠神经炎症的影响。方法:随机选择15只健康大鼠为健康组(尾静脉注射生理盐水),BM模型大鼠随机分为BM组(尾静脉注射生理盐水)、L-Hyp组(尾静脉注射10mg/kg Hyp)、H-Hyp组(尾静脉注射50mg/kg Hyp)、TXNIP-AAV组(尾静脉注射TXNIP-AAV)、AAV组(尾静脉注射AAV-NC)、白藜芦醇(Res)组(尾静脉注射30mg/kg Res)。采用Loeffler评分评估大鼠治疗后神经功能;血细胞分析仪测定白细胞(WBC)数量;酶联免疫吸附法分析脑脊液活性氧(ROS)、白细胞介素(IL)-1β、IL-6、肿瘤坏死因子α(TNF-α)水平;测定各组大鼠脑组织含水量;HE染色观察脑组织病理学变化;TUNEL染色观察脑组织细胞凋亡;免疫印迹法分析脑组织中TXNIP、NLRP3、caspase-1、凋亡相关斑点蛋白(ASC)、离子钙结合适配分子1(Iba1)、IL-1β蛋白表达。结果:与健康组比,BM组脑脊液ROS、WBC、IL-1β、IL-6、TNF-α水平及脑组织含水量、细胞凋亡率增加,Loeffler评分减少(P<0.05);与BM组比,L-Hyp组、H-Hyp组Loeffler评分增加,脑脊液ROS、WBC、IL-1β、IL-6、TNF-α水平及脑组织含水量、细胞凋亡率减少(P<0.05);与H-Hyp组比,TXNIP-AAV组、AAV组脑脊液ROS、WBC、IL-1β、IL-6、TNF-α水平及脑组织含水量、细胞凋亡率增加,Loeffler评分减少,Res组Loeffler评分增加,脑脊液ROS、WBC、IL-1β、IL-6、TNF-α水平及脑组织含水量、细胞凋亡率减少(P<0.05)。健康组大鼠脑组织形态正常;BM组、TXNIP-AAV组、AAV组脑组织形态改变,细胞排列散乱,出现核皱缩;L-Hyp组、H-Hyp组、Res组脑组织形态有所改善,细胞坏死、皱缩减少。与健康组比,BM组脑组织中TXNIP、NLRP3、caspase-1、ASC、Iba1、IL-1β表达增加(P<0.05);与BM组比,LHyp组、H-Hyp组脑组织中TXNIP、NLRP3、caspase-1、ASC、Iba1、IL-1β表达减少(P<0.05);与H-Hyp组比,TXNIP-AAV组、AAV组脑组织中TXNIP、NLRP3、caspase-1、ASC、Iba1、IL-1β表达增加,Res组脑组织中TXNIP、NLRP3、caspase-1、ASC、Iba1、IL-1β表达减少(P<0.05)。结论:Hyp可能抑制BM大鼠神经炎症,其机制可能与抑制TXNIP/NLRP3通路有关。
Objective:To analyze the effects of Hypericin(Hyp)on neuroinflammation in bacterial meningitis(BM)rats by regulating the thioredoxin-interacting protein(TXNIP)/nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)signaling pathway.Methods:Fifteen healthy rats were randomly selected as the control group(tail vein injection of saline).BM model rats were randomly divided into BM group(tail vein injection of saline),L-Hyp group(tail vein injection of 10mg/kg Hyp),H-Hyp group(tail vein injection of 50mg/kg Hyp),TXNIP-AAV group(tail vein injection of TXNIP-AAV),AAV group(tail vein injection of AAV-NC),and Resveratrol(Res)group(tail vein injection of 30mg/kg Res).Neurological function after treatment was evaluated using the Loeffler score;white blood cell(WBC)count was determined using a blood cell analyzer;reactive oxygen species(ROS),interleukin(IL)-1β,IL-6,and tumor necrosis factor-alpha(TNF-α)levels in cerebrospinal fluid(CSF)were analyzed by ELISA;brain tissue water content was measured;pathological changes in brain tissue were observed using HE staining;cell apoptosis in brain tissue was observed using TUNEL staining;and protein expressions of TXNIP,NLRP3,caspase-1,apoptosis-associated speck-like protein(ASC),ionized calcium-binding adaptor molecule 1(Iba1),and IL-1βin brain tissue were analyzed using Western blotting.Results:Compared with the control group,the BM group showed increased levels of ROS,WBC,IL-1β,IL-6,and TNF-αin CSF,increased brain tissue water content and cell apoptosis rate,and decreased Loeffler scores(P<0.05).Compared with the BM group,the L-Hyp and H-Hyp groups showed increased Loeffler scores,and decreased levels of ROS,WBC,IL-1β,IL-6,and TNF-αin CSF,brain tissue water content,and cell apoptosis rate(P<0.05).Compared with the H-Hyp group,the TXNIP-AAV and AAV groups showed increased levels of ROS,WBC,IL-1β,IL-6,and TNF-αin CSF,brain tissue water content,and cell apoptosis rate,and decreased Loeffler scores(P<0.05),whereas the Res group showed increased Loeffler scores and decreased levels of ROS,WBC,IL-1β,IL-6,and TNF-αin CSF,brain tissue water content,and cell apoptosis rate(P<0.05).The brain tissues of rats in the control group showed normal morphology;in the BM,TXNIP-AAV,and AAV groups,brain tissues showed morphological changes,disordered cell arrangement,and nuclear shrinkage;in the L-Hyp,H-Hyp,and Res groups,brain tissues showed improved morphology,with reduced cell necrosis and shrinkage.Compared with the control group,the BM group showed increased expressions of TXNIP,NLRP3,caspase-1,ASC,Iba1,and IL-1βin brain tissues(P<0.05).Compared with the BM group,the LHyp and H-Hyp groups showed decreased expressions of TXNIP,NLRP3,caspase-1,ASC,Iba1,and IL-1βin brain tissues(P<0.05).Compared with the H-Hyp group,the TXNIP-AAV and AAV groups showed increased expressions of TXNIP,NLRP3,caspase-1,ASC,Iba1,and IL-1βin brain tissues,while the Res group showed decreased expressions of these proteins(P<0.05).Conclusion:Hypericin may inhibit neuroinflammation in BM rats,possibly through inhibition of the TXNIP/NLRP3 pathway.
作者
陈杰
姜慧琳
王丽艳
杨俊杰
CHEN Jie;JIANG Huilin(Union Jiangbei Hospital,Huazhong University of Science and Technology,Hubei Wuhan 430100,China)
出处
《河北医学》
CAS
2024年第8期1261-1267,共7页
Hebei Medicine
基金
湖北省武汉市卫健委中医药科研项目,(编号:WZ22C18)。