亚慢性氟中毒大鼠脑组织RAGE/p38MAPK/NF-κB信号通路的变化及银杏叶提取物和RAGE阻断剂的保护作用  

Changes of RAGE/p38MAPK/NF-κB signaling pathway in brain tissue of subchronic fluorosis rats and protective effects of Ginkgo biloba extract and RAGE antagonist

作　　者：梁秋哲 李红梅 邓婕[2] 张婷[2] 马彦琳 张凯琳 LIANG Qiuzhe;LI Hongmei;DENG Jie;ZHANG Ting;MA Yanlin;ZHANG Kailin(School of Basic Medicine,Guizhou Medical University,Guiyang,Guizhou 550004,China;Ministry of Education Key Laboratory of Endemic and Minority Diseases/Key Laboratory of Molecular Biology of Guizhou Province,Guizhou Medical University,Guiyang,Guizhou 550004,China)

机构地区：[1]贵州医科大学基础医学院,贵州贵阳550004 [2]贵州医科大学地方病与少数民族性疾病教育部重点实验室/贵州省分子生物学重点实验室,贵州贵阳550004

出　　处：《环境与职业医学》2023年第5期577-582,共6页Journal of Environmental and Occupational Medicine

基　　金：国家自然科学基金项目(81860562、U1812403);贵州省科技厅基金项目(黔科合基础[2019]1276、黔科合基础-ZK[2021]一般490、黔科合基础-ZK[2022]一般401)。

摘　　要：[背景]长期摄入过量的氟会蓄积在脑组织中并造成神经损伤和学习记忆能力的衰退,晚期糖基化终产物受体(RAGE)/p38丝裂原活化蛋白激酶(p38MAPK)/核因子κB(NF-κB)信号通路是其中的重要机制。[目的]研究亚慢性氟中毒大鼠脑组织RAGE/p38MAPK/NF-κB信号通路的改变,并探讨银杏叶提取物(EGb761)和RAGE阻断剂(FPS-ZM1)对神经记忆能力的保护作用。[方法]选择雄性清洁级SD大鼠90只,将其分为9组,每组10只,造模周期6个月。分别为对照组(C组):自由饮用自来水(含氟量<0.5 mg·L^(−1)),低、高剂量染氟组(LF、HF组):分别自由饮用含氟量为10、50 mg·L^(−1)的自来水;银杏叶提取物干预组(CE、LFE、HFE组):在C、LF、HF组的基础上,每天给予100 mg·kg^(−1)·d^(−1)EGb761灌胃;FPS-ZM1干预组(CF、LFF、HFF组):在C、LF、HF组的基础上,造模结束前7 d每天腹腔注射1 mg·kg^(−1)·d^(−1)FPS-ZM1。检测各组大鼠的脑氟和血氟含量;水迷宫实验检测各组学习记忆能力;尼氏染色检测各组大鼠海马的病理形态学改变;蛋白免疫印迹法检测脑组织中RAGE及其配体高迁移率族蛋白B1(HMGB1)、NF-κB、p38MAPK、磷酸化p38MAPK(p-p38MAPK)、白介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)蛋白的表达水平;实时荧光定量PCR法检测RAGE、HMGB1、p38MAPK的mRNA表达水平。[结果]与C组血氟、脑氟相比,LF、HF组大鼠的血氟、脑氟含量均增加(P均<0.05)。水迷宫实验结果显示,与C组相比,LF组、HF组的逃避潜伏时间延长,穿越次数减少;与HF组相比,HFE组和HFF组的逃避潜伏时间缩短,穿越次数增加(P均<0.05)。尼氏染色结果显示,与C组相比,HF组尼氏体数量减少;与HF组相比,HFE组和HFF组尼氏体数量增加。Western blotting结果显示:与C组RAGE、HMGB1、NF-κB、p38MAPK、p-p38MAPK、IL-6、TNF-α蛋白相对表达量相比,LF组RAGE、HMGB1、NF-κB、p-p38MAPK、IL-6和HF组的RAGE、HMGB1、NF-κB、p38MAPK、p-p38MAPK、IL-6、TNF-α[Background]Fluorine accumulates in the brain tissue after long-term excessive intake and subsequently cause nerve damage and decline of learning and memory ability.Receptor of advanced glycation end-products(RAGE)/p38 mitogen-activated protein kinase(p38MAPK)/nuclear factor kappa-B(NF-κB)signaling pathway is considered to be involved in the associated mechanism.[Objective]To study the changes of RAGE/p38MAPK/NF-κB signaling pathway in rats with subchronic fluorosis,and to explore the protective effects of extract of Ginkgo biloba 761(EGb761)and RAGE antagonist(FPS-ZM1)on neuromemory ability.[Methods]Ninety male clean SD rats were divided into 9 groups with 10 rats in each group.The modeling period was 6 months.Control group(C group):free drinking tap water(fluoride content<0.5 mg·L^(−1)),low-and high-dose fluoride groups(LF group,HF group):free drinking tap water with 10 or 50 mg·L^(−1)fluoride;intervention group of Ginkgo biloba extract(CE,LFE,and HFE groups):on the basis of the C group,LF group,and HF group,100 mg·kg^(−1)·d^(−1)EGb761 was given daily via intragastric administration;FPS-ZM1 intervention groups(CF,LFF,and HFF groups):7 d before the end of modeling,1 mg·kg^(−1)·d^(−1)FPS-ZM1 was injected intraperitoneally daily on the basis of the C group,LF group,and HF group.The contents of fluoride in brain and blood of each group were detected.The learning and memory ability was tested by water maze experiment.The histopathologic changes of the hippocampus were detected by Nissl staining.The protein expression levels of RAGE and its ligand high mobility group protein B1(HMGB1),NF-κB,p38MAPK,phospho-p38MAPK(p-p38MAPK),interleukin-6(IL-6),and tumour necrosis factor-α(TNF-α)in brain tissue were detected by Western blotting.The mRNA expression levels of RAGE,HMGB1,and p38MAPK were detected by quantitative real-time PCR.[Results]Compared with the C group,the contents of blood fluoride and brain fluoride in the LF and the HF groups were increased(P<0.05).The results of the water maze experime

关 键 词：亚慢性氟中毒 脑组织 晚期糖基化终产物受体 银杏叶提取物 阻断剂 

分 类 号：R11[医药卫生—公共卫生与预防医学]