摘要
目的探讨青蒿琥酯(Artesunate,ART)调节白介素-6(Interleukin-6,IL-6)/信号转导及转录激活因子3(Signal transduction and transcription activating factor 3,STAT3)信号通路对神经病理性疼痛(Neuropathic pain,NP)大鼠模型的治疗作用。方法随机选取12只大鼠记为正常对照(Normal control,NC)组,其他大鼠构建NP大鼠模型,造模成功大鼠随机平分为NP组、ART组(25 mg/kg的ART)、IL-6组(0.075μg/kg的IL-6重组蛋白)、ART+IL-6组(25 mg/kg的ART+0.075μg/kg的IL-6重组蛋白),每组各12只,药物连续处理1周;NC组和NP组给予等量的生理盐水;24 h后进行后续实验,检测机械缩足反射阈值(Mechanical withdrawal threshold,MWT)以及热痛阈(Thermal withdrawal latency,TWL);酶联免疫吸附测定(Enzyme linked immunosorbent assay,ELISA)法检测肿瘤坏死因子-α(Tumor necrosis factor-α,TNF-α)、IL-1β水平;苏木精-伊红(Hematoxylin-eosin,HE)染色观察背根神经节病理变化;免疫荧光染色检测小胶质细胞和星形胶质细胞活性;Western blot检测IL-6/STAT3信号通路相关蛋白水平。结果NC组大鼠背根神经节细胞排列整齐,细胞间距规则,细胞核大小正常;与NC组比较,NP组背根神经节细胞排列紊乱,细胞间距较大,细胞核较小,可见凋亡细胞,大鼠白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)水平、离子钙接头蛋白分子1(Ion calcium junction protein molecule 1,Iba1)、胶质纤维酸性蛋白(Glial fibrillary acidic protein,GFAP)阳性细胞数、IL-6,STAT3蛋白水平显著升高(P<0.05),MWT值、TWL值显著下降(P<0.05);与NP组比较,ART组背根神经节细胞的排列较规则,细胞核略小,MWT值、TWL值显著增高(P<0.05),IL-1β,TNF-α水平、Iba1,GFAP阳性细胞数、IL-6,STAT3蛋白水平显著降低(P<0.05),而IL-6重组蛋白与ART的作用相反,即IL-6重组蛋白消除了ART对大鼠NP的治疗作用。结论ART可能通过抑制IL-6/STAT3信号通路来减轻NP大鼠模型的疼痛。
Objective To investigate the therapeutic effect of artesunate(ART)on neuropathic pain(NP)in rats through the regulation of the interleukin-6(IL-6)/signal transduction and transcription activating factor 3(STAT3)signaling pathway.Methods Twelve rats were randomly selected as the normal control(NC)group,while the remaining rats were used to construct the NP rat model.The successfully generated rats were then randomly separated into four groups:the NP group,ART group(25 mg/kg ART),IL-6 group(0.075μg/kg IL-6 recombinant protein),and ART+IL-6 group(25 mg/kg ART+0.075μg/kg IL-6 recombinant protein),with 12 rats in each group.The rats in each group were continuously treated with the indicated drugs for 1 week,while the NC and NP groups were given equal amounts of physiological saline.Subsequent experiments were conducted after 24 hours.The mechanical withdrawal threshold(MWT)and thermal withdrawal latency(TWL)were measured.The levels of interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)were detected using the enzyme-linked immunosorbent assay(ELISA)method.Pathological changes in the dorsal root ganglion were observed using hematoxylin-eosin(HE)staining.The activity of microglia and astrocytes was detected using immunofluorescence staining.The levels of IL-6/STAT3 signaling pathway-related proteins were measured using Western blotting.Results The dorsal root ganglion cells of rats in the NC group were neatly arranged with regular cell spacing and standardized nuclear size.In contrast,the dorsal root ganglion cells in the NP group exhibited a disorderly arrangement,greater cell spacing,and fewer nuclei.Apoptotic cells were also observed.The levels of TNF-αand IL-1β,as well as the numbers of ion calcium junction protein molecule 1(Iba1)-and glial fibrillary acidic protein(GFAP)-positive cells,were significantly increased in the rats,as were the levels of the IL-6 and STAT3 proteins(P<0.05).The MWT and TWL were significantly lower(P<0.05).However,in the ART group,the arrangement of dorsal root ganglion cells was more regular,with slightly fewer nuclei.The MWT and TWL were significantly greater(P<0.05).Moreover,the levels of IL-1βand TNF-α,the numbers of Iba1-and GFAP-positive cells,and the levels of the IL-6 and STAT3 proteins were significantly lower in the rats than in the control rats(P<0.05).Interestingly,the effect of the recombinant IL-6 protein on ART was opposite;it eliminated the therapeutic effect of ART on rat NPs.Conclusion ART may alleviate pain in NP rats by inhibiting the IL-6/STAT3 signaling pathway.
作者
李先超
罗红玉
Li Xianchao;Luo Hongyu(Department of Pain,Huanggang Central Hospital Affiliated to Changjiang University,Huanggang Hubei 438000;不详)
出处
《卒中与神经疾病》
2024年第2期187-191,205,共6页
Stroke and Nervous Diseases
基金
湖北省卫生健康委2021-2022年度卫生健康科研立项项目(WJ2021Q016)。
作者简介
通信作者:罗红玉。
