Identification and Quantification of Total Coumarins from Urtica dentata Hand and Its Roles in Promoting Immune Tolerance via TLR4-Mediated Dendritic Cell Immaturation

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  • XIANG Ming
    Department of Pharmacology, School of Pharmacy, Tongji Medical College, Huazhong University of Science and Technology
  • LU Jingli
    Department of Pharmacology, School of Pharmacy, Tongji Medical College, Huazhong University of Science and Technology
  • ZHANG Chengliang
    Department of Pharmaceutics, Tongji Hospital, Huazhong University of Science and Technology
  • LAN Yan
    Department of Pharmacology, School of Pharmacy, Tongji Medical College, Huazhong University of Science and Technology
  • ZHOU Hong
    Department of Pharmacology, School of Pharmacy, Tongji Medical College, Huazhong University of Science and Technology
  • LI Xiping
    Department of Pharmaceutics, Tongji Hospital, Huazhong University of Science and Technology
  • PENG Wen
    Department of Geriatrics, Union Hospital, Huazhong University of Science and Technology

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  • Identification and Quantification of Total Coumarins from <i>Urtica dentata</i> Hand and Its Roles in Promoting Immune Tolerance <i>via</i> TLR4-Mediated Dendritic Cell Immaturation

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Urtica dentata Hand (UDH) is traditionally used in the Alpine region as a herbal medicine. Immunotherapy using total coumarins (TC) of UDH has been proposed, yet the cellular and molecular mechanisms remain incompletely characterized. Additionally, there is no method available for the quantification of the main coumarins in UDH. We describe maturation-resistant, TC-conditioned dendritic cell (DC), which expressed much lower MHC class II (I-Ak) and CD86, showed reduced capacity to stimulate effector T cell responses and upregulated PD-Ll (programmed death ligand-1). TC-DC-stimulated regulatory cells (Treg) were superior alloantigen-specific suppressor of the T effector response as compared to those stimulated by control (CTR)-DC. Furthermore, TC-conditioned DC increased the levels of Foxp3 and CTLA-4 in the CD25 T cell population. TC-DC downregulated toll like receptor 4 (TLR4) protein expression in response to LPS. This indicates that down-regulation of TLR4 in response to TC on DC is a critical signaling pathway that regulates the phenotype and function of DC. We also established a sensitive and specific high-performance liquid chromatography-diodearray detection-mass spectrometry (HPLC-DAD-MS) method for simultaneous identification of its main coumarins, 6,6',7,7'-tetramethoxyl-8,8'-biscoumarin (1), 7,7'-dihydroxy-6,6'-dimethoxy-8,8'-biscoumarin (2), 7,7'-dimethoxy-6,6'-biscoumarin (3), and scoparone (4). A demonstration of this mechanism and the method for identification and quantification of TC in UDH endorsed their potential as a tolerance-promoting herbal medicine to prevent or treat transplantation rejection and autoimmune diseases.

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