脾交感神经调控DRG巨噬细胞浸润和极化在小鼠糖尿病神经病理性痛中的作用  

Role of splenic sympathetic nerve-regulated infiltration and polarization of dorsal root ganglion macrophages in diabetic neuropathic pain in mice

作　　者：韩守孟 何万友[2] 陈欣[3] 吴范灿 梁洪宾 王龙[1] 王汉兵[2] Han Shoumeng;He Wanyou;Chen Xin;Wu Fancan;Liang Hongbin;Wang Long;Wang Hanbing(Department of Anesthesiology,Renmin Hospital of Wuhan University,Wuhan 430060,China;Department of Anesthesiology,The First People′s Hospital of Foshan,Foshan 528000,China;Department of Anesthesiology,Shaanxi Provincial People′s Hospital,Xi′an 710068,China)

机构地区：[1]武汉大学人民医院麻醉科,武汉430060 [2]佛山市第一人民医院麻醉科,佛山528000 [3]陕西省人民医院麻醉科,西安710068

出　　处：《中华麻醉学杂志》2025年第1期71-76,共6页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金(82201549)。

摘　　要：目的评价脾交感神经调控背根神经节(DRG)巨噬细胞浸润和极化在小鼠糖尿病神经病理性痛(DNP)中的作用。方法选取SPF级健康雄性C57BL/6J小鼠48只,6周龄,体质量20~22 g,采用随机数字表法分为4组(n=12):对照组(Con组)、DNP组、DNP+假手术组(DNP+Sham组)和DNP+脾交感神经去除术组(DNP+SS组)。DNP+SS组采用6-羟基多巴胺溶液毁损脾脏交感神经,DNP+Sham组用0.2%抗坏血酸盐溶液代替6-羟基多巴胺溶液,术后小鼠休养2周。于小鼠8周龄时腹腔注射链脲佐菌素120 mg/kg制备糖尿病模型。分别于造模前1 d和造模后第7、14、21、28天时测定机械缩足反应阈(MWT)。最后一次行为学测试后,麻醉后取DRG,采用免疫荧光法检测巨噬细胞标志物离子钙结合适配器分子1(Iba1)、降钙素基因相关肽(CGRP)和肿瘤坏死因子α(TNF-α)的表达;采用qRT-PCR法检测巨噬细胞M1型标志物CD16、TNF-α、诱导型一氧化氮合酶(iNOS)和M2型标志物白细胞介素-10(IL-10)、转化生长因子β1(TGF-β1)、CD206的mRNA表达。结果与Con组相比,DNP组造模后第14、21及28天时MWT降低,脊髓DRG CGRP和TNF-α表达上调,Iba1阳性细胞计数升高,CD16、TNF-α、iNOS的mRNA表达上调(P<0.05),IL-10、TGF-β1、CD206的mRNA表达差异无统计学意义(P>0.05);与DNP组和DNP+Sham组相比,DNP+SS组造模后第14、21及28天时MWT升高,脊髓DRG CGRP和TNF-α表达下调,Iba1阳性细胞计数降低,CD16、TNF-α、iNOS的mRNA表达下调,IL-10、TGF-β1、CD206的mRNA表达上调(P<0.05);DNP组与DNP+Sham组各时点上述各指标比较差异无统计学意义(P>0.05)。结论脾交感神经激活可促进DRG巨噬细胞浸润与极化,从而参与小鼠DNP的过程。ObjectiveTo evaluate the role of splenic sympathetic nerve-regulated infiltration and polarization of dorsal root ganglion(DRG)macrophages in diabetic neuropathic pain(DNP)in mice.MethodsForty-eight specific pathogen-free male C57BL/6 mice,aged 6 weeks,weighing 20-22 g,were divided into 4 groups(n=12 each)using a random number table method:control group(Con group),DNP group,DNP plus sham operation group(DNP+Sham group),and diabetes mellitus induced by DNP plus splenic sympathetic denervation group(DNP+SS group).In DNP+SS group,the splenic sympathetic denervation procedures were performed using 6-hydroxydopamine solution,while a solvent of 0.2%ascorbic acid saline solution was used as a substitute for 6-hydroxydopamine solution in DNP+Sham group.After a two-week recovery,the diabetes mellitus was induced by intraperitoneal injection of streptozotocin 120 mg/kg in mice at 8 weeks of age.The mechanical paw withdrawal threshold(MWT)were measured on day 1 before developing the model and on days 7,14,21 and 28 after developing the model.After the last behavioral testing,the DRG was taken after anesthesia for determination of the expression of the macrophage marker ionized calcium-binding adaptor molecule 1(Iba1),calcitonin gene-related peptide(CGRP),and tumor necrosis factor-α(TNF-α)(by immunofluorescence)and expression of M1 phenotype markers(CD16,TNF-α,inducible nitric oxide synthase[iNOS])and M2 phenotype markers(interleukin-10[IL-10],transforming growth factor-β1[TGF-β1],and CD206)mRNA(using quantitative real-time polymerase chain reaction).ResultsCompared with Con group,the MWT was significantly decreased on days 14,21 and 28 after developing the model,the expression of CGRP and TNF-αin the DRG was up-regulated,the count of Iba1-positive cells was increased,the expression of CD16,TNF-αand iNOS mRNA was up-regulated(P<0.05),and no significant change was found in the expression of IL-10,TGF-β1 and CD206 in DNP group(P>0.05).Compared with DNP group and DNP+Sham group,the MWT was significantly increased on day

关 键 词：糖尿病神经病变 交感神经系统 背根神经节 巨噬细胞 极化 

分 类 号：R614[医药卫生—麻醉学]