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基于脊髓中枢敏化的镇痛筛选方法挖掘延胡索的抗神经病理性疼痛成分。

Based on spinal central sensitization creating analgesic screening approach to excavate anti-neuropathic pain ingredients of Corydalis yanhusuo W.T.Wang.

机构信息

Jiangsu Key Laboratory of TCM Evaluation and Translational Research, School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing, Jiangsu, 211198, China.

Jiangsu Key Laboratory of TCM Evaluation and Translational Research, School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing, Jiangsu, 211198, China; State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing, Jiangsu, 210009, China.

出版信息

J Ethnopharmacol. 2024 Jan 30;319(Pt 1):117084. doi: 10.1016/j.jep.2023.117084. Epub 2023 Sep 2.

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE

Corydalis Rhizome (RC) as a traditional analgesic Chinese medicine is the dried tuber of Corydalis yanhusuo W.T.Wang. Many efforts have revealed that RC could effectively alleviate neuropathic pain, while its active ingredients in neuropathic pain are still not clear.

AIM OF THE STUDY

Spinal central sensitization contributes greatly to neuropathic pain, and neuron, astrocyte and microglia play important roles in spinal central sensitization. The aim of the present study is to excavate active compounds in RC regulating spinal central sensitization to inhibit neuropathic pain.

MATERIALS AND METHODS

Immunofluorescence and western blotting were used to determine protein expression levels. Gene expression levels were detected by RT-PCR. PC12 neuronal cells, C6 astrocyte cells, and BV2 microglia cells were cultured for in vitro studies. Targeting multi types of cells extraction combined with HPLC-Q-TOF-MS/MS was established to identify components binding to above cells. Animal studies were used to verify the analgesic activities of components.

RESULTS

Total alkaloids of RC (RC-TA) significantly relieved neuropathic pain in chronic constriction injury (CCI) rats and repressed spinal central sensitization. Eight components of RC-TA were found to bind to PC12, C6, or BV2 cells. They could respectively suppress the activation of cells in vitro and alleviate CCI-induced neuropathic pain, among which glaucine and dehydrocorydaline induced antinociception was stronger than l-THP. Meanwhile, glaucine had no effect on acute or chronic inflammatory pain, and its antinociception in neuropathic pain could be abolished by dopamine D1 receptor agonist.

CONCLUSIONS

Employing multi types of cells based on spinal central sensitization rather than single cell may allow for more thorough excavation of active substances. Glaucine was firstly found could attenuate neuropathic pain but not other types of pain which indicated that different alkaloids in RC exert distinct analgesic effects on different pain models, and gluacine has the potential to be developed as an analgesic drug specifically for neuropathic pain relieving.

摘要

民族药理学相关性

延胡索(RC)作为一种传统的镇痛中药,是紫堇属植物延胡索的干燥根茎。大量研究表明,RC 能有效缓解神经性疼痛,但其在神经性疼痛中的活性成分仍不清楚。

研究目的

脊髓中枢敏化对神经性疼痛有很大贡献,神经元、星形胶质细胞和小胶质细胞在脊髓中枢敏化中发挥重要作用。本研究旨在挖掘 RC 中调节脊髓中枢敏化以抑制神经性疼痛的活性化合物。

材料和方法

采用免疫荧光和 Western blot 测定蛋白表达水平,RT-PCR 检测基因表达水平。体外培养 PC12 神经元细胞、C6 星形胶质细胞和 BV2 小胶质细胞进行研究。建立靶向多种细胞提取方法结合 HPLC-Q-TOF-MS/MS 鉴定与上述细胞结合的成分。动物研究用于验证成分的镇痛活性。

结果

RC 总生物碱(RC-TA)显著缓解慢性缩窄性损伤(CCI)大鼠神经性疼痛,并抑制脊髓中枢敏化。从 RC-TA 中发现 8 种成分与 PC12、C6 或 BV2 细胞结合。它们分别在体外抑制细胞的激活,并缓解 CCI 诱导的神经性疼痛,其中,格尔碱和脱氢紫堇碱的镇痛作用强于 l-THP。同时,格尔碱对急性或慢性炎症性疼痛没有影响,其在神经性疼痛中的镇痛作用可被多巴胺 D1 受体激动剂阻断。

结论

采用基于脊髓中枢敏化的多种细胞而非单一细胞的方法,可能更彻底地挖掘活性物质。首次发现格尔碱可减轻神经性疼痛,但不能减轻其他类型的疼痛,这表明 RC 中的不同生物碱对不同的疼痛模型有不同的镇痛作用,格尔碱有可能被开发为专门用于缓解神经性疼痛的镇痛药。

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