肠道菌群缺失在紫杉醇诱导的神经病理性疼痛中的作用  

Mechanism of gut microbiota deletion in regulating neuropathic pain induced by paclitaxel

作　　者：巴茜远 郝悦 蒋昌宇 廖宇良 BA Xiyuan;HAO Yue;JIANG Changyu;LIAO Yuliang(Key Laboratory of Pain Department,Huazhong University of Science and Technology Union Shenzhen Hospital,Shenzhen,Guangdong 518060,China;不详)

机构地区：[1]深圳市华中科技大学协和深圳医院疼痛科重点实验室,深圳广东518060 [2]深圳大学医学部基础医学院药学系

出　　处：《中国微生态学杂志》2024年第12期1382-1387,共6页Chinese Journal of Microecology

基　　金：国家自然科学基金(81971065)。

摘　　要：目的探索肠道菌群缺失对紫杉醇诱导的神经病理性疼痛行为的影响及相关机制。方法32只C57雄性小鼠随机分为4组(n=8):对照组(Control组)、抗生素组(ASC组)、紫杉醇组(PTX组)和抗生素+紫杉醇组(PTX+ASC组)。紫杉醇造模前,抗生素组小鼠口服ASC三联广谱抗生素(1 mg/mL),持续2周。第1、3、5、7天腹腔注射紫杉醇(2 mg/kg)建立神经病理性疼痛模型。紫杉醇给药前和给药后第14天分别测定小鼠的热缩足潜伏期(TWL)和机械缩足反应阈(MWT)。行为学实验结束后,取小鼠粪便样本(5~6粒/只)用于16S rRNA扩增子测序分析。Western blot法检测小鼠脊髓水平Toll样受体4(TLR4)蛋白和核因子κB(NF-κB)蛋白的表达情况。免疫荧光检测脊髓星形胶质细胞的活化情况。ELISA实验检测脊髓内肿瘤坏死因子α(TNF-α)和白细胞介素-1β(IL-1β)的表达水平。结果与对照组相比,口服ASC抗生素的小鼠,肠道菌群在科水平上Muribaculaceae、乳杆菌科和阿克曼菌科相对丰度显著下降(P<0.05);紫杉醇2 mg/kg造模后,小鼠机械/热痛阈值显著下降(P<0.05),小鼠脊髓TLR4和NF-κB蛋白和促炎性因子TNF-α、IL-1β表达水平显著升高(P<0.05),脊髓星形胶质细胞异常活化(P<0.05)。与紫杉醇组相比,抗生素+紫杉醇组小鼠的机械/热痛阈值显著升高(P<0.05)。小鼠脊髓内TLR4和NF-κB蛋白和TNF-α、IL-1β的表达显著降低(P<0.05),星形胶质细胞活化水平显著减弱(P<0.05)。结论ASC抗生素三联给药造成的肠道菌群缺失可缓解紫杉醇诱导的小鼠神经病理性疼痛,并可抑制紫杉醇诱导的脊髓TLR4/NF-κB信号通路蛋白的过表达,星形胶质细胞活化和炎性反应。Objective To explore the impact of gut microbiota deficiency on paclitaxel-induced neuropathic pain and its related mechanisms.Methods Thirty-two C57 male mice were randomly divided into 4 groups(n=8):control group(C group),antibiotic group(ASC group),paclitaxel group(PTX group),and antibiotic+paclitaxel group(PTX+ASC group).The mice in the ASC group were orally administered three broad-spectrum antibiotics(1 mg/mL)for 2 weeks.The neuropathic pain model was established using intraperitoneal injection of paclitaxel(2 mg/kg)on the 1st,3rd,5th and 7th day.The thermal withdraw latency(TWL)and mechanical withdraw threshold(MWT)of mice were measured on day 0 and 14,respectively.After the behavior test,the fecal samples(5~6 grain of feces/mouse)were taken for 16S rDNA amplification sequencing analysis.The expression of toll-like receptor 4(TLR4)and nuclear factor kappa-B(NF-κB)protein in mice spinal cord was detected with Western blot.The activation of astrocytes in spinal cord was detected with immunofluores-cence.ELISA method was used to detect the levels of inflammatory factors including TNF-αand IL-1β.Results Compared with the control group,mice orally treated with ASC antibiotics showed a significant decrease in the abundance of Muribaculaceae,Lactobacillaceae and Akkermansiaceae at the family level(P<0.05);after the administration of paclitaxel(2 mg/kg),the mechanical/thermal pain threshold of mice significantly decreased(P<0.05),the expression levels of TLR4 and NF-κB proteins and pro-inflammatory factors TNF-αand IL-1βin the spinal cord of mice significantly increased(P<0.05),and spinal astrocytes were abnormally activated(P<0.05).Compared with the paclitaxel group,the mechanical/thermal pain threshold of mice in the antibiotic+paclitaxel group significantly increased(P<0.05).The expressions of TLR4 and NF-κB proteins,as well as TNF-αand IL-1βin the spinal cord of mice significantly reduced(P<0.05),and the activation level of astrocytes was significantly weakened(P<0.05).Conclusion The neuropathic pain

关 键 词：神经病理性疼痛 肠道菌群 星形胶质细胞 TOLL样受体4 

分 类 号：R338.2[医药卫生—人体生理学]