肠道菌群对大鼠紫杉醇神经病理性疼痛模型的影响  

Effects of gut microbiome on paclitaxel-induced neuropathic pain in rats

作　　者：朱彬 郑彩虹 边英麟 施海滨 ZHU Bin;ZHENG Cai-hong;Bian Ying-lin;SHI Hai-bin(Department of Anesthesiology,Hangzhou Women's Hospital,Hangzhou 310000,China;Department of Anesthesiology,the Affiliated Hangzhou First People's Hospital,Zhejiang University School of Medicine,Hangzhou 310000,China)

机构地区：[1]杭州市妇产科医院,浙江杭州310000 [2]浙江大学医学院附属杭州市第一人民医院,浙江杭州310000

出　　处：《中国病理生理杂志》2022年第11期1990-1997,共8页Chinese Journal of Pathophysiology

基　　金：浙江省医药卫生科技项目(No.2020KY231,No.2022KY943)。

摘　　要：目的:探讨肠道菌群对大鼠紫杉醇(PTX)神经病理性疼痛模型的影响。方法:构建大鼠PTX神经病理性疼痛模型,并通过喂养大鼠广谱抗生素或移植粪便改变大鼠肠道菌群;SD大鼠随机分为对照组、PTX+生理盐水组(PTX组)、PTX+抗生素(Abx)组和PTX+粪菌移植(FMT)组,每组6只。测定大鼠的机械缩足反射阈值(MWT)和热缩足潜伏期(TWL);16S rDNA分析肠道菌群种属;Western blot检测各组结肠组织Toll样受体4(TLR4)及丝裂原活化蛋白激酶(MAPKs)[包括磷酸化p38 MAPK(p-p38 MAPK)、磷酸化细胞外信号调节激酶1/2(p-ERK1/2)和磷酸化c-Jun氨基末端激酶(p-JNK)]的表达蛋白水平。PTX模型大鼠造模第1天鞘内分别注射TLR4激动剂LPS-PG Ultrapure、TLR4抑制剂TAK242和p38抑制剂SB203580(每组6只),检测大鼠的MWT和TWL,ELISA法检测各组大鼠血清白细胞介素1β(IL-1β)和肿瘤坏死因子α(TNF-α)含量。结果:(1)PTX组大鼠MWT和TWL随着造模时间的增加呈下降趋势,造模第21天最敏感,均低于对照组(P<0.01);PTX造模后第12天,PTX+FMT组MWT和TWL较PTX组呈上升趋势(P<0.05);PTX组与PTX+Abx组比较无显著差异。(2)16S rDNA分析结果显示,各组粪便菌群种属存在显著差异(P<0.05)。(3)Western blot实验结果显示,PTX组与PTX+Abx组TLR4、p-p38、p-JNK和p-ERK1/2均高于对照组(P<0.05)。(4)PTX大鼠模型鞘内注射TLR4或p38抑制剂,均可缓解大鼠MWT和TWL下降趋势(P<0.05)。(5)ELISA结果显示,PTX诱导的神经病理性疼痛大鼠模型中血清炎症因子IL-1β和TNF-α水平显著高于对照组(P<0.01);PTX模型大鼠鞘内注射LPS-PG Ultrapure促进IL-1β和TNF-α的释放(P<0.01);鞘内注射TAK242或SB203580均抑制IL-1β和TNF-α的释放(P<0.01)。结论:肠道菌群可能通过激活TLR4/MAPKs信号通路介导PTX大鼠模型神经病理性疼痛。AIM:To investigate the effect of gut microbiome on paclitaxel(PTX)-induced neuropathic pain(PINP)in rats.METHODS:The PINP models were established in Sprague-Dawley(SD)rats,and the gut microbiome of rats was altered by feeding broad-spectrum antibiotics or transplanting fecal microbiota.The SD rats were randomly divided into 4 groups of 6 each,namely control group,PTX group,PTX+antibiotic(Abx)group,and PTX+fecal microbiota transplantation(FMT)group.The mechanical withdrawal threshold(MWT)and thermal withdrawal latency(TWL)of rat hind paws were tested.Additionally,microbial analyses of the stool were performed using 16S rDNA sequencing.The protein levels of Toll-like receptor 4(TLR4)and mitogen-activated protein kinases(MAPKs),including phosphorylated p38 MAPK(p-p38 MAPK),phosphorylated extracellular signal-regulated kinase 1/2(p-ERK1/2)and phosphorylated c-Jun N-terminal kinase(p-JNK),in the colon tissues were determined by Western blot.The PINP rat models from the 3 groups were intrathecally injected with TLR4 agonist LPS-PG Ultrapure,TLR4 inhibitor TAK242,or p38 inhibitor SB203580,and then the MWT and TWL of rat hind paws were detected.Lastly,the content of inflammatory cytokines interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)in the colon tissues was detected ELISA.RESULTS:The MWT and TWL in PTX group were lower than those in control group(P<0.01)and were the lowest on day 21.On day 12 after PTX treatment,the MWT and TWL in PTX+FMT group were higher than those in PTX group(P<0.05).However,there was no statistically significant difference between PTX and PTX+Abx groups.In contrast,16S rDNA analysis of the stools revealed significant differences in the gut microbiome among all groups(P<0.05).Additionally,the protein levels of TLR4,pp38,p-JNK and p-ERK1/2 in PTX and PTX+Abx groups were higher than those in control group(P<0.05).Moreover,the levels of IL-1β and TNF-αin PTX group were higher than those in control group(P<0.01).However,intrathecal injection of TAK242 or SB203580 relieved mechanical and t

关 键 词：肠道微生物 丝裂原活化蛋白激酶 神经病理性疼痛 紫杉醇 TOLL样受体4 

分 类 号：R741.02[医药卫生—神经病学与精神病学] R363.2[医药卫生—临床医学]