Analysis of the regulatory effect of Angelica dahurica on the MrgprD-TRPA1 pathway in neuropathic pain
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1.Department of Basic Medicine,School of Health and Nursing,Wuxi Taihu University;2.School of Medicine,Yangzhou University

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    Abstract:

    Objective Neuropathic pain was a kind of chronic pain caused by central or peripheral nervous system injury, dysfunction or transient disorder. Its mechanism was related to some receptors. In order to analyze the analgesic effect of Angelica dahurica in neuropathic pain and its regulation on MrgprD-TRPA1 signaling pathway. Methods Firstly, the CCI mouse model was prepared by using sterile surgical ligation and wrapping of the sciatic nerve in 30 mice. Secondly, the VonFrey experiment was used to detect the behavioral changes in pain induced by mechanical stimulation in mice, and the thermal radiation experiment was used to evaluate the thermal hyperalgesia of Angelica dahurica in mice. Then, the effects of Angelica dahurica on the expression of MrgprD and TRPA1 proteins, the number of DRG positive neurons, and the mRNA levels of MrgprD and TRPA1 in mice were detected by Western blot, immunofluorescence, and RT-PCR, respectively. Finally, the differences in fluorescence signal intensity were analyzed through calcium imaging experiments on HEK293 cells after single transfection and co-transfection of MrgprD and TRPA1 plasmids, respectively. Results A total of 25 CCI mouse models were successfully prepared, with a modeling rate of 83.33% (25/30). After the 7th day, CCI mice showed the most significant differences in mechanical stimulation threshold and thermal radiation foot contraction latency, and reached the lowest value.The mechanical threshold and foot retraction latency of CCI mice treated with Angelica dahurica were significantly higher than those treated with pure water and CCI mice not treated with Angelica dahurica (P<0.05). The expression levels of MrgprD and TRPA1 proteins in CCI mice treated with Angelica dahurica were significantly lower than those in CCI mice treated with pure water (0.73 ± 0.11 vs 2.69 ± 0.23, 0.42 ± 0.09 vs 2.03 ± 0.18, P<0.05). The number of MrgprD and TRPA1 positive neurons in DRG of CCI mice treated with Angelica dahurica was significantly lower than that of CCI mice treated with pure water (654 ± 47 vs 1162 ± 63, P<0.05). The relative expression levels of MrgprD and TRPA1 mRNA in CCI mice treated with Angelica dahurica were significantly lower than those in CCI mice treated with pure water and CCI mice not treated with Angelica dahurica(P<0.05). The fluorescence intensity in HEK293 cells co transfected with MrgprD and TRPA1 plasmids was significantly higher than that in single transfected and blank controls (P<0.05). Conclusion This study successfully established a mouse CCI model through ligation and winding, exploring the analgesic effect and mechanism of Angelica dahurica in the CCI model, and proving that MrgprD-TRPA1 is an important target for neuropathic pain. Angelica dahurica can inhibit the degree of neuropathic pain by regulating the signal transduction pathway of MrgprD-TRPA1, which laid a foundation for further research on the development of new clinical analgesic drugs and analgesic mechanisms.

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History
  • Received:September 21,2023
  • Revised:January 15,2024
  • Adopted:January 15,2024
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