摘要
目的:探讨番茄红素调节Janus激酶2(JAK2)/信号转导和转录激活因子3(STAT3)/血管内皮生长因子(VEGF)信号通路对脑小血管病(CSVD)大鼠血脑屏障和神经损伤的影响。方法:通过同种系微栓子体外注入法制备50只CSVD大鼠模型,随机分为模型组、番茄红素低剂量(65 mg/kg)组、番茄红素高剂量(85 mg/kg)组、AG490(JAK2抑制剂,3.5 mg/kg)组、番茄红素高剂量(85 mg/kg)+AG490(3.5 mg/kg)组,每组10只,另选取10只SD大鼠作为假手术组,以番茄红素和AG490分组处理后,采用Morris水迷宫实验和避暗实验检测大鼠认知能力;伊文思蓝法检测大鼠血脑屏障通透性;尼氏染色检测大鼠海马神经元数量;试剂盒检测大鼠脑组织炎症因子前列腺素E2(PGE2)、肿瘤坏死因子-α(TNF-α)及氧化应激因子过氧化氢酶(CAT)、超氧化物歧化酶(SOD)、丙二醛(MDA)水平;免疫印迹法检测大鼠脑组织基质金属蛋白酶(MMP)2、MMP9、紧密连接相关蛋白(ZO-1、Occludin)及JAK2/STAT3/VEGF通路相关蛋白表达。结果:与假手术组比较,模型组大鼠穿台次数、目标象限停留时间、步入潜伏期、海马神经元数量、脑组织CAT与SOD水平、ZO-1与Occludin、VEGF蛋白表达、p-JAK2/JAK2、p-STAT3/STAT3显著降低(P<0.05),错误次数、伊文思蓝含量、脑组织PGE2、TNF-α、MDA水平及MMP2、MMP9蛋白表达显著升高(P<0.05)。与模型组比较,番茄红素低剂量组、番茄红素高剂量组大鼠穿台次数、目标象限停留时间、步入潜伏期、海马神经元数量、脑组织CAT与SOD水平、ZO-1与Occludin、VEGF蛋白表达、p-JAK2/JAK2、p-STAT3/STAT3均升高(P<0.05),错误次数、伊文思蓝含量、脑组织PGE2、TNF-α、MDA水平及MMP2、MMP9蛋白表达均降低(P<0.05),且高剂量番茄红素作用更强;AG490组大鼠各指标变化趋势与番茄红素各剂量组相反,且AG490可逆转番茄红素的作用。结论:番茄红素可通过激活JAK2/STAT3/VEGF信号通路抑制CSVD大鼠炎症与氧化应激,从而减轻其血脑屏障和神经损伤,并提升其认知能力。
Objective:To investigate the effect of lycopene on blood-brain barrier and nerve damage in rats with cerebral small vessel disease(CSVD)by regulating Janus kinase 2(JAK2)/signal transducer and activator of transcription 3(STAT3)/vascular endo-thelial growth factor(VEGF)signaling pathway.Methods:Fifty CSVD rat models were prepared by in vitro injection of the same germ-line microemboli,and were randomly grouped into model group,low-dose lycopene(65 mg/kg)group,high-dose lycopene(85 mg/kg)group,AG490(JAK2 inhibitor,3.5 mg/kg)group,and high-dose lycopene(85 mg/kg)+AG490(3.5 mg/kg)group,with 10 cases in each group,another 10 SD rats were regarded as sham operation group.After grouping with lycopene and AG490,cognitive ability of rats was detected by Morris water maze test and dark avoidance test;permeability of blood-brain barrier in rats was detected by Evans blue method;the number of rat hippocampal neurons was detected by Nissl staining;levels of inflammatory factors prostaglandin E2(PGE2),tumor necrosis factor-α(TNF-α),and oxidative stress factors catalase(CAT),superoxide dismutase(SOD),malondialde-hyde(MDA)in rats brain tissue were detected by kits;expressions of matrix metalloproteinase(MMP)2,MMP9,tight junction-related proteins(ZO-1,Occludin)and JAK2/STAT3/VEGF pathway-related proteins in rats brain were detected by Western blotting.Results:Compared with sham operation group,the times of rats crossing the platform,the time of staying in the target quadrant,step through latency,the number of hippocampal neurons,levels of CAT and SOD in brain tissue,protein expressions of ZO-1,Occludin and VEGF,and p-JAK2/JAK2,p-STAT3/STAT3 were obviously decreased in model group(P<0.05),while the error times,Evans blue content,levels of brain tissue PGE2,TNF-αand MDA,and protein expressions of MMP2 and MMP9 were obviously increased(P<0.05).Compared with model group,the times of rats crossing the platform,the time of staying in the target quadrant,step through la-tency,the number of hippocampal neurons,levels of CAT and SOD in brain tissue,protein expressions of ZO-1,Occludin and VEGF,and p-JAK2/JAK2,p-STAT3/STAT3 were all increased in low-dose lycopene group and high-dose lycopene group(P<0.05),while the error times,Evans blue content,levels of brain tissue PGE2,TNF-αand MDA,and protein expressions of MMP2 and MMP9 were all decreased(P<0.05),and high dose lycopene was more effective;the change trend of each index in AG490 group was opposite to that in lycopene groups,and AG490 could reverse the effect of lycopene.Conclusion:Lycopene can inhibit inflammation and oxidative stress in CSVD rats by activating JAK2/STAT3/VEGF signaling pathway,thereby reducing the blood-brain barrier and nerve damage,and improving their cognitive ability.
作者
刘传芬
宋征宇
张晓丽
胡瑾
LIU Chuanfen;SONG Zhengyu;ZHANG Xiaoli;HU Jin(Hebei North University,Zhangjiakou 075000,China;Third Department of Neurology,the First Affiliated Hospital of Hebei North University,Zhangjiakou 075000,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2024年第11期2343-2349,共7页
Chinese Journal of Immunology
基金
河北省中医药管理局资助项目(2023100)。
作者简介
刘传芬,女,硕士,住院医师,主要从事脑小血管病方面的研究,E-mail:liuchuanfen1993f@163.com;通信作者:胡瑾,女,硕士,主任医师,主要从事神经系统疾病学研究,E-mail:miofs47@163.com。
