Effects of Jiawei Longdan Xiegan Decoction on Neuroinflammation in Postherpetic Neuralgia Rats via the cAMP/PKA/PPARγ Pathway
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摘要:
目的 探讨加味龙胆泻肝汤（ jiawei longdan xiegan decoction，JWD）对带状疱疹后遗神经痛（postherpetic neuralgia，PHN）大鼠的镇痛作用及其对脊髓神经炎症的影响，并探究其是否通过调控环磷酸腺苷/蛋白激酶A/过氧化物酶体增殖物激活受体γ（cyclic adenosine monophosphate/protein kinase A/peroxisome proliferator-activated receptor γ，cAMP/PKA/PPARγ）信号通路发挥作用。 方法 采用坐骨神经慢性压迫性损伤联合角叉菜胶局部致炎法建立PHN大鼠模型，将造模成功大鼠随机分为模型组（Model）、JWD低剂量组（JWD-L）、高剂量组（JWD-H）、加巴喷丁阳性对照组（GBP）、高剂量+cAMP激活剂FSK组（JWD-H+FSK），另设假手术组（Sham），n = 10/组。连续给药28 d后，检测机械痛阈（paw withdrawal mechanical threshold，PWMT）与热痛阈（paw withdrawal thermal latency，PWTL），观察自发痛及焦虑/抑郁样行为；采用酶联免疫吸附实验检测脊髓中cAMP、PKA及炎症因子，白细胞介素-1β（interleukin-1β，IL-1β）、白细胞介素-6（interleukin-6，IL-6）、肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）含量；免疫荧光染色观察小胶质细胞与星形胶质细胞活化；蛋白质免疫印迹检测（phosphorylated cAMP response element-binding protein/cAMP response element-binding protein，p-CREB/CREB）、PPARγ、磷酸化核因子-κB（phosphorylated nuclear factor-κB，p-NF-κB）、NOD样受体家族含吡喃结构域蛋白3（NOD-like receptor family pyrin domain-containing protein 3，NLRP3）、环氧合酶2（cyclooxygenase-2，COX-2）、诱导型一氧化氮合酶（inducible nitric oxide synthase，iNOS）以及脑源性神经营养因子（brain-derived neurotrophic factor，BDNF）、酪氨酸激酶B（tyrosine kinase B，TrkB）、胶质细胞源性神经营养因子（glial cell line-derived neurotrophic factor，GDNF）、重排转染期间原癌基因（rearranged during transfection，RET）、突触后致密蛋白 95（postsynaptic density protein 95，PSD-95）等蛋白表达。脊髓HE与尼氏染色评估神经元损伤及存活率，检测背根神经节（dorsal root ganglion，DRG）中炎症因子IL-1β、IL-6、TNF-α及瞬时受体电位香草酸亚型1（transient receptor potential vanilloid 1，TRPV1）蛋白表达。 结果 与Model相比，JWD-H组可显著提高PWMT和PWTL，减少自发痛行为，改善焦虑与抑郁样行为（P < 0.01）。同时，JWD-H能显著抑制脊髓背角小胶质细胞和星形胶质细胞活化，降低IL-1β、IL-6、TNF-α水平，下调cAMP含量及p-CREB/CREB比值，上调PPARγ蛋白表达，并抑制NF-κB通路活化（P < 0.001）。此外，JWD-H在抑制疼痛相关BDNF/TrkB表达的同时，上调了神经保护性GDNF/RET及突触蛋白PSD-95的表达（P < 0.001）。病理学结果显示，JWD-H可显著减轻脊髓背角神经元固缩、空洞形成，提高Nissl染色神经元存活率（P < 0.001）；同时降低DRG中炎症细胞浸润，下调IL-1β、IL-6、TNF-α含量及TRPV1蛋白表达（P < 0.001）。而JWD-H和GBP在部分指标（COX-2和iNOS、PKA、PPARγ、BDNF、TrkB、RET、IL-6、TNF-α、星形胶质细胞荧光强度等）上存在显著差异（P < 0.05），JWD-L组仅在部分指标上有改善作用（P < 0.05）。JWD-H的上述作用在联合cAMP激活剂FSK后被部分逆转（P < 0.05）。 结论 JWD能有效缓解PHN大鼠的神经病理性疼痛及相关情绪障碍，其机制可能与调控cAMP/PKA/PPARγ信号轴，抑制脊髓神经炎症、NF-κB通路活化，并促进神经保护与修复有关。 -
关键词:
- 加味龙胆泻肝汤 /
- 带状疱疹后遗神经痛 /
- cAMP/PKA/PPARγ通路 /
- 神经炎症
Abstract:Objective To investigate the analgesic effect of Jiawei Longdan Xiegan Decoction （JWD） on rats with postherpetic neuralgia （PHN） and its influence on spinal neuroinflammation, and to explore whether its effect is mediated by regulating the cyclic adenosine monophosphate/protein kinase A/peroxisome proliferator-activated receptor γ （cAMP/PKA/PPARγ） signaling pathway. Methods A PHN rat model was established by chronic constriction injury of the sciatic nerve combined with local inflammation induced by carrageenan. Successfully modeled rats were randomly divided into model group （Model）, low-dose JWD group （JWD-L）, high-dose JWD group （JWD-H）, gabapentin positive control group （GBP）, and high-dose JWD plus cAMP activator forskolin group （JWD-H+FSK）, with a sham surgery group （Sham） also included, n = 10 per group. After 28 days of continuous administration, the paw withdrawal mechanical threshold （PWMT） and paw withdrawal thermal latency （PWTL） were detected, and spontaneous pain as well as anxiety- and depression-like behaviors were observed. Enzyme-linked immunosorbent assay was used to detect the levels of cAMP, PKA and inflammatory cytokines including interleukin-1β （IL-1β）, interleukin-6 （IL-6） and tumor necrosis factor-α （TNF-α） in the spinal cord. Immunofluorescence staining was performed to observe the activation of microglia and astrocytes. Western blot was used to detect the expression of phosphorylated cAMP response element-binding protein/cAMP response element-binding protein （p-CREB/CREB）, PPARγ, phosphorylated nuclear factor-κB （p-NF-κB）, NOD-like receptor family pyrin domain-containing protein 3 （NLRP3）, cyclooxygenase-2 （COX-2）, inducible nitric oxide synthase （iNOS）, as well as brain-derived neurotrophic factor （BDNF）, tyrosine kinase B （TrkB）, glial cell line-derived neurotrophic factor （GDNF）, rearranged during transfection （RET） and postsynaptic density protein 95 （PSD-95）. Hematoxylin-eosin （HE） and Nissl staining of the spinal cord were used to evaluate neuronal injury and survival rate. The levels of inflammatory cytokines IL-1β, IL-6, TNF-α and the protein expression of transient receptor potential vanilloid 1 （TRPV1） in dorsal root ganglion （DRG） were determined. Results Compared with the Model group, JWD-H group significantly increased PWMT and PWTL, reduced spontaneous pain behaviors, and ameliorated anxiety- and depression-like behaviors （P < 0.01）. Meanwhile, JWD-H markedly inhibited activation of microglia and astrocytes in the spinal dorsal horn, decreased the levels of IL-1β, IL-6 and TNF-α, reduced cAMP content and the p-CREB/CREB ratio, upregulated PPARγ protein expression, and suppressed the activation of the NF-κB pathway （P < 0.001）. In addition, while inhibiting pain-related BDNF/TrkB expression, JWD-H upregulated the expression of neuroprotective GDNF/RET and the synaptic protein PSD-95 （P < 0.001）. Pathology results showed that JWD-H obviously alleviated neuronal pyknosis and cavitation in the spinal dorsal horn and increased the neuronal survival rate shown by Nissl staining （P < 0.001）. Meanwhile, JWD-H reduced inflammatory cell infiltration in DRG and downregulated the levels of IL-1β, IL-6, TNF-α as well as TRPV1 protein expression （P < 0.001）. Significant differences were observed between JWD-H and GBP in certain indicators （including COX-2 and iNOS, PKA, PPARγ, BDNF, TrkB, RET, IL-6, TNF-α, and astrocyte fluorescence intensity, etc.; P < 0.05）, while the JWD-L group showed improvement only in some indicators （P < 0.05）. The above effects of JWD-H were partially reversed when combined with the cAMP activator FSK （P < 0.05）. Conclusion JWD effectively alleviates neuropathic pain and associated emotional disorders in PHN rats, possibly through modulating the cAMP/PKA/PPARγ signaling axis, inhibiting spinal neuroinflammation and NF-κB pathway activation, and promoting neuroprotection and repair. -
图 1 JWD对PHN大鼠痛觉过敏的影响（n = 10）
A：各组大鼠不同时间点PWMT比较；B：各组大鼠不同时间点PWTL比较；*P < 0.05；**P < 0.01；***P < 0.001。
Figure 1. Effects of JWD on hyperalgesia in PHN rats （n = 10）
图 2 JWD对PHN大鼠自发痛及情绪行为的影响（n = 10）
A：各组大鼠自发舔足次数比较；B：各组大鼠抬足次数比较；C：各组大鼠旷场中央区停留时间比较；D：各组大鼠强迫游泳不动时间比较；*P < 0.05；**P < 0.01；***P < 0.001。
Figure 2. Effects of JWD on spontaneous pain and emotional behavior in PHN rats （n = 10）
图 3 JWD对PHN大鼠脊髓背角病理损伤和DRG炎症因子及TRPV1表达的影响（n = 10）
A：各组脊髓背角HE染色和Nissl染色代表性图片（比例尺：50 μm）；B：神经元存活率柱状图；C～D：DRG中TRPV1蛋白条带图及定量分析结果；E：IL-1β含量；F：IL-6含量；G：TNF-α含量；*P < 0.05；**P < 0.01；***P < 0.001。
Figure 3. Effects of JWD on pathological injury of the spinal dorsal horn and the expression of inflammatory cytokines and TRPV1 in the DRG of PHN rats （n = 10）
图 4 JWD对PHN大鼠脊髓神经炎症的影响 （n = 10）
A：各组代表性大鼠脊髓背角小胶质细胞和星形胶质细胞免疫荧光图像（比例尺：50 μm）；B～C：免疫荧光定量分析结果；D：TNF-α含量；E：IL-6含量；F：IL-1β含量；*P < 0.05；**P < 0.01；***P < 0.001。
Figure 4. Effect of JWD on spinal cord neuroinflammation in PHN rats （n = 10）
图 5 JWD对PHN大鼠脊髓神经营养因子及突触蛋白表达的影响（n = 10）
A：各组大鼠代表性蛋白表达条带图；B～F：BDNF、TrkB、GDNF、RET、PSD-95的蛋白表达定量分析柱状图；*P < 0.05；***P < 0.001。
Figure 5. Effect of JWD on the expression of spinal neurotrophic factors and synaptic proteins in PHN rats （n = 10）
图 6 JWD对PHN大鼠脊髓cAMP/PKA/PPARγ通路关键分子表达的影响（n = 10）
A：各组大鼠cAMP含量；B：各组大鼠PKA含量；C：各组大鼠代表性蛋白条带图；D～E：蛋白表达的定量分析柱状图；*P < 0.05；**P < 0.01；***P < 0.001。
Figure 6. Effect of JWD on the expression of key molecules in the spinal cord cAMP/PKA/PPARγ pathway in PHN rats （n = 10）
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[1] Tang J, Zhang Y, Liu C, et al. Therapeutic strategies for postherpetic neuralgia: Mechanisms, treatments, and perspectives[J]. Curr Pain Headache Rep, 2023, 27(9): 307-319. doi: 10.1007/s11916-023-01146-x [2] Shen Y, Lin P. The role of cytokines in postherpetic neuralgia[J]. J Integr Neurosci, 2025, 24(4): 25829. doi: 10.31083/JIN25829 [3] Chen S C, Chen Y H, Song Y, et al. Upregulation of phosphodiesterase 7A contributes to concurrent pain and depression via inhibition of cAMP-PKA-CREB-BDNF signaling and neuroinflammation in the hippocampus of mice[J]. Int J Neuropsychopharmacol, 2024, 27(10): pyae040. doi: 10.1093/ijnp/pyae040 [4] Zhai Q, Wang K, Zhang D, et al. Perampanel ameliorates nitroglycerin-induced migraine through inhibition of the cAMP/PKA/CREB signaling pathway in the trigeminal ganglion in rats[J]. Korean J Pain, 2023, 36(3): 335-346. doi: 10.3344/kjp.23039 [5] Sokołowska P, Bleibel L, Owczarek J, et al. PPARγ, NF-κB and the UPR pathway as new molecular targets in the anti-inflammatory actions of NSAIDs: Novel applications in cancers and central nervous system diseases?[J]. Adv Clin Exp Med, 2024, 33(9): 1007-1022. doi: 10.17219/acem/174243 [6] Fu X X, Wei B, Cao H M, et al. Telmisartan alleviates Alzheimer’s disease-related neuropathologies and cognitive impairments[J]. J Alzheimers Dis, 2023, 94(3): 919-933. doi: 10.3233/JAD-230133 [7] Ju Y, Li L, Zhao Y, et al. Prostaglandin E2 inhibits the differentiation of T regulatory cells by Peroxisome Proliferator-Activated Receptor-Gamma during allergic rhinitis[J]. Asian Pac J Allergy Immunol, 2025, 43(2): 214-226. doi: 10.12932/ap-210923-1695 [8] 乐春云, 陈翠香, 王方智. 加味龙胆泻肝汤结合中医外治法治疗带状疱疹的临床观察[J]. 中国民间疗法, 2021, 29(22): 58-60. doi: 10.19621/j.cnki.11-3555/r.2021.2222 [9] 杨悦, 刘秭慧. 龙胆泻肝汤治疗带状疱疹临床疗效的 Meta 分析[J]. 湖南中医杂志, 2021, 37(7): 136-140. doi: 10.16808/j.cnki.issn1003-7705.2021.07.049 [10] 朱英, 曾涛, 王昌文, 等. 加味龙胆泻肝汤联合疏肝调神针刺法治疗带状疱疹后神经痛肝经郁热证临床观察及对细胞免疫、白细胞介素35、CX3C趋化因子受体1的影响[J]. 河北中医, 2025, 47(7): 1084-1088+1093. doi: 10.3969/j.issn.1002-2619.2025.07.007 [11] Lu C C, Lin C Y, Lu Y Y, et al. CDDO regulates central and peripheral sensitization to attenuate post-herpetic neuralgia by targeting TRPV1/PKC-δ/p-Akt signals[J]. J Cellular Molecular Medi, 2024, 28(6): e18131. doi: 10.1111/jcmm.18131 [12] Liu S L, Zhang H E, Kang J Y, et al. Spinal cord stimulation alleviates sensitization of neuropathic pain by upregulating G protein-coupled receptors to inhibit overexpression of Cav2.2 and its downstream excitatory neurotransmitters[J]. J Pain Res, 2025, 18: 2775-2790. doi: 10.2147/JPR.S514719 [13] Forouzanfar F, Tanha N K, Pourbagher-Shahri A M, et al. Synergistic effect of ellagic acid and gabapentin in a rat model of neuropathic pain[J]. Metab Brain Dis, 2023, 38(4): 1421-1432. doi: 10.1007/s11011-023-01190-x [14] 吴春燕, 杜鹃. 中医治疗带状疱疹后遗神经痛的现状及展望[J]. 中国病原生物学杂志, 2025, 20(11): 1519-1521. doi: 10.13350/j.cjpb.251129 [15] 李力翠, 杨祎峰, 毛伟, 等. 龙胆草提取物液体敷料治疗面部敏感性皮肤的疗效及对皮肤屏障功能的影响[J]. 中国美容医学, 2023, 32(11): 98-102. doi: 10.15909/j.cnki.cn61-1347/r.006291 [16] 刘靖丽, 张拴, 张光辉, 等. 黄芩苷和栀子苷抗炎活性和抗炎机理的理论研究[J]. 化学研究与应用, 2023, 35(3): 669-673. doi: 10.3969/j.issn.1004-1656.2023.03.025 [17] 秦万里. 中医治疗带状疱疹知识图谱构建、挖掘与临床研究[D]. 北京: 北京中医药大学, 2024. [18] 韩文彪, 韩文娟, 时宏琛, 等. 短时程脊髓电刺激和背根神经节电刺激治疗带状疱疹相关神经痛和改善睡眠质量的临床疗效[J]. 昆明医科大学学报, 2025, 46(12): 112-122. doi: 10.12259/j.issn.2095-610X.S20251213 [19] Hou J, Wang X, Zhang J, et al. Chuanxiong Renshen decoction inhibits Alzheimer’s disease neuroinflammation by regulating PPARγ/NF-κB pathway[J]. Drug Des Devel Ther, 2024, 18: 3209-3232. doi: 10.2147/DDDT.S462266 [20] Wei X, Wang L, Hua J, et al. Inhibiting BDNF/TrkB. T1 receptor improves resiniferatoxin-induced postherpetic neuralgia through decreasing ASIC3 signaling in dorsal root ganglia[J]. J Neuroinflammation, 2021, 18(1): 96. doi: 10.1186/s12974-021-02148-5 [21] 刘书画, 李艺, 胡小莉, 等. 胶质细胞源性神经营养因子在脊髓损伤中的相关作用研究[J]. 赣南医学院学报, 2021, 41(9): 965-970. doi: 10.3969/j.issn.1001-5779.2021.09.019 -
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