摘要
目的:探究瞬时感受器电位香草酸亚型1(transient receptor potential vanilloid 1,TRPV1)介导小胶质细胞自噬在带状疱疹后神经疼痛中的作用机制。方法:40只小鼠随机分为正常组(Con)、带状疱疹后神经痛组(postherpetic neuralgia,PHN)、阴性对照组(PHN-sh-NC)、TRPV1干扰组(PHN-sh-TRPV1),每组10只。检测小鼠机械缩足阈值(mechanical withdrawal thresh-old,MWT)和热痛阈潜伏期(thermal withdrawal latency,TWL);ELISA检测血清中肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)、白细胞介素-1β(interleukin-1 beta,IL-1β)、白细胞介素6(interleukin-6,IL-6)、脑源性神经营养因子(brain-derived neuro-trophic factor,BDNF)水平;透射电镜检测脊髓组织中自噬体形成;免疫荧光检测脊髓组织中微管相关蛋白轻链3(microtubule-associated protein 1 light chain 3,LC3)、离子钙结合适配器分子1(ionized calcium-binding adapter molecule 1,Iba-1)表达;Western blot检测脊髓组织中Beclin-1、自噬微管相关蛋白轻链3β(microtubule-associated protein 1 light chain 3 beta,LC3B)、类似核孔糖蛋白p62(similar to nuclear pore glycoprotein p62,p62)蛋白表达。结果:和Con组相比,PHN组和PHN-sh-NC组小鼠、血清BNDF(t=10.49,P<0.001)水平及脊髓组织中p62(P=0.004)蛋白表达明显降低,小鼠血清中TNF-α(t=26.27,P<0.001)、IL-1β(t=17.0,P<0.001)、IL-6(t=25.48,P<0.001)水平及脊髓组织中Iba-1(P=0.002)、LC3(P<0.001)表达和Beclin-1(P=0.001)、LC3B(P<0.001)蛋白表达明显升高;和PHN组相比,PHN-sh-TRPV1组小鼠MWT、血清BNDF(t=5.174,P<0.001)水平及脊髓组织中p62(P<0.001)蛋白表达明显升高;TWL、血清中TNF-α(t=20.57,P<0.001)、IL-1β(t=8.260,P<0.001)和IL-6(t=19.81,P<0.001)水平及脊髓组织中Iba-1(P<0.001)、LC3(P=0.001)表达和LC3B(P<0.001)、Beclin-1(P<0.001)蛋白表达明显降低。结论:PHN小鼠脊髓小胶质细胞自噬激活,抑制TRPV1表达可通过抑制小胶质细胞自噬缓解带状疱疹后神经痛小鼠的疼痛。
Objective:To investigate the mechanism of transient receptor potential vanilloid subfamily 1(TRPV1)-mediated microglia autophagy in postherpetic neuralgia.Methods:Forty mice were randomly divided into control group,postherpetic neuralgia(PHN)group,PHN-sh-NC group,and PHN-sh-TRPV1 group,with 10 mice in each group.We tested the mice’s mechanical withdrawal threshold(MWT)and thermal withdrawal latency(TWL);measured serum levels of tumor necrosis factor-alpha(TNF-α),interleukin(IL)-1β,IL-6,and brain-derived neurotrophic factor(BDNF)by enzyme-linked immunosorbent assay;assessed the formation of au-tophagosomes in the spinal cord tissues by transmission electron microscopy;measured the expression of microtubule-associated pro-tein 1 light chain 3(LC3)and ionized calcium binding adaptor molecule 1(Iba-1)in the spinal cord tissues by immunofluorescence assay;and determined the protein expression of Beclin-1,microtubule-associated protein 1 light chain 3B(LC3B),and p62 in the spinal cord tissues by Western blot.Results:Compared with the control group,the PHN group and PHN-sh-NC group had sig-nificant decreases in MWT,serum BNDF level(t=10.49,P<0.001),and p62(P=0.004)protein expression in the spinal cord tissues and significant increases in TWL,serum TNF-α(t=26.27,P<0.001),IL-1β(t=17.0,P<0.001),and IL-6 levels(t=25.48,P<0.001),and the expression of Iba-1(P=0.002),LC3(P<0.001),LC3B(P=0.001),and Beclin-1 proteins(P=0.001)in the spinal cord tissues.Compared with the PHN group,the PHN-sh-TRPV1 group had a significantly higher MWT,a significantly higher serum BNDF level(t=5.174,P<0.001,a significantly higher p62 protein expression level(P<0.001)in the spinal cord tissues,a significantly lower TWL,significantly lower serum TNF-α(t=20.57,P<0.001),IL-1β(t=8.260,P<0.001),and IL-6 levels(t=19.81,P<0.001),and signifi-cantly lower expression levels of Iba-1(P<0.001),LC3(P<0.001),LC3B(P=0.001),and Beclin-1(P<0.001)in the spinal cord tis-sues.Conclusion:Microglia autophagy is activated in the spinal cord of PHN mice,and suppressing the expression of TRPV1 can inhibit microglia autophagy to relieve pain in PHN mice.
作者
陈太长
蹇忠禄
丁维
陈荣
陈英
吴闵
唐松江
贾敏
Chen Taichang;Jian Zhongu;Ding Wei;Chen Rong;Chen Ying;Wu Min;Tang Songjiang;Jia Min(Department of Anesthesiology,The First Affiliated Hospital of Guizhou University of Traditional Chinese Medicine;Department of Urology,The First Affiliated Hospital of Guizhou University of Traditional Chinese Medicine;Department of Traditional Chinese Medicine Internal Medicine,The First Clinical Medical College of Traditional Chinese Medicine,Guizhou University of Traditional Chinese Medicine;Department of Dermatology,The First Affiliated Hospital of Guizhou University of Traditional Chinese Medicine)
出处
《重庆医科大学学报》
北大核心
2025年第8期1058-1063,共6页
Journal of Chongqing Medical University
基金
2024年度省卫生健康委科学技术基金资助项目(编号:2024GZWJKJXM1026)。
