Potent Antagonists of RORγt, Cardenolides from Calotropis gigantea, Exhibit Discrepant Effects on the Differentiation of T Lymphocyte Subsets. 2019

Juan Liu, and Li-Ping Bai, and Fen Yang, and Xiaojun Yao, and Kawai Lei, and Christopher Wai Kei Lam, and Qibiao Wu, and Yuxin Zhuang, and Riping Xiao, and Kangsheng Liao, and Hioha Kuok, and Ting Li, and Liang Liu
State Key Laboratory of Quality Research in Chinese Medicines, Macau Institute for Applied Research in Medicine and Health , Macau University of Science and Technology , Avenida Wai Long , Taipa , Macau 999078 , People's Republic of China.

RORγt is the master transcription factor of IL-17 cytokine expression and Th17 lymphocyte differentiation, which are responsible for the induction of many autoimmune diseases. Recently, RORγt has become an attractive target for drug development to treat these types of diseases, and the field of RORγt antagonist research is now extremely competitive. In our current study, molecular docking was applied to demonstrate that cardenolides, including uscharin, calactin, and calotropin derived from Calotropis gigantea, probably directly bind to RORγt. Therefore, the inhibitory effect was further validated using a luciferase reporter assay. Because RORγt is the key transcriptional factor for Th17 differentiation, the effects of these compounds on Th17 differentiation were studied by flow cytometry. The results showed that uscharin, calactin, and calotropin inhibited Th17 differentiation from 100 to 500 nM. Furthermore, uscharin had a better effect than digoxin, a well-known inverse agonist of RORγt, in reducing Th17 polarization. Additionally, the effects of the cardenolides on the differentiation of other Th lineages, including Th1, Th2, and Treg, were investigated. Uscharin suppressed Th1, Th2, and Treg cell differentiation, while calactin suppressed the differentiation of Th1 cells, and calotropin did not influence the other T cell subsets, indicating that calactin suppressed Th1 and Th17 differentiation, and calotropin selectively quenched Th17 polarization. Structural analysis of the three compounds showed that the selectivity of uscharin, calactin, and calotropin on the suppression of the different subsets of T cells is correlated to the minor differences in their chemical structures. Collectively, calactin and calotropin have greater potential to be developed as lead compounds than uscharin to treat autoimmune diseases mediated by Th17 and/or Th1 cells.

UI MeSH Term Description Entries
D008810 Mice, Inbred C57BL One of the first INBRED MOUSE STRAINS to be sequenced. This strain is commonly used as genetic background for transgenic mouse models. Refractory to many tumors, this strain is also preferred model for studying role of genetic variations in development of diseases. Mice, C57BL,Mouse, C57BL,Mouse, Inbred C57BL,C57BL Mice,C57BL Mice, Inbred,C57BL Mouse,C57BL Mouse, Inbred,Inbred C57BL Mice,Inbred C57BL Mouse
D002298 Cardenolides C(23)-steroids with methyl groups at C-10 and C-13 and a five-membered lactone at C-17. They are aglycone constituents of CARDIAC GLYCOSIDES and must have at least one double bond in the molecule. The class includes cardadienolides and cardatrienolides. Members include DIGITOXIN and DIGOXIN and their derivatives and the STROPHANTHINS. Cardenolide
D002454 Cell Differentiation Progressive restriction of the developmental potential and increasing specialization of function that leads to the formation of specialized cells, tissues, and organs. Differentiation, Cell,Cell Differentiations,Differentiations, Cell
D005434 Flow Cytometry Technique using an instrument system for making, processing, and displaying one or more measurements on individual cells obtained from a cell suspension. Cells are usually stained with one or more fluorescent dyes specific to cell components of interest, e.g., DNA, and fluorescence of each cell is measured as it rapidly transverses the excitation beam (laser or mercury arc lamp). Fluorescence provides a quantitative measure of various biochemical and biophysical properties of the cell, as well as a basis for cell sorting. Other measurable optical parameters include light absorption and light scattering, the latter being applicable to the measurement of cell size, shape, density, granularity, and stain uptake. Cytofluorometry, Flow,Cytometry, Flow,Flow Microfluorimetry,Fluorescence-Activated Cell Sorting,Microfluorometry, Flow,Cell Sorting, Fluorescence-Activated,Cell Sortings, Fluorescence-Activated,Cytofluorometries, Flow,Cytometries, Flow,Flow Cytofluorometries,Flow Cytofluorometry,Flow Cytometries,Flow Microfluorometries,Flow Microfluorometry,Fluorescence Activated Cell Sorting,Fluorescence-Activated Cell Sortings,Microfluorimetry, Flow,Microfluorometries, Flow,Sorting, Fluorescence-Activated Cell,Sortings, Fluorescence-Activated Cell
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D015153 Blotting, Western Identification of proteins or peptides that have been electrophoretically separated by blot transferring from the electrophoresis gel to strips of nitrocellulose paper, followed by labeling with antibody probes. Immunoblotting, Western,Western Blotting,Western Immunoblotting,Blot, Western,Immunoblot, Western,Western Blot,Western Immunoblot,Blots, Western,Blottings, Western,Immunoblots, Western,Immunoblottings, Western,Western Blots,Western Blottings,Western Immunoblots,Western Immunoblottings
D016176 T-Lymphocyte Subsets A classification of T-lymphocytes, especially into helper/inducer, suppressor/effector, and cytotoxic subsets, based on structurally or functionally different populations of cells. T-Cell Subset,T-Cell Subsets,T-Lymphocyte Subset,Subset, T-Cell,Subset, T-Lymphocyte,Subsets, T-Cell,Subsets, T-Lymphocyte,T Cell Subset,T Cell Subsets,T Lymphocyte Subset,T Lymphocyte Subsets
D057809 HEK293 Cells A cell line generated from human embryonic kidney cells that were transformed with human adenovirus type 5. 293T Cells,HEK 293 Cell Line,HEK 293 Cells,Human Embryonic Kidney Cell Line 293,Human Kidney Cell Line 293,293 Cell, HEK,293 Cells, HEK,293T Cell,Cell, 293T,Cell, HEK 293,Cell, HEK293,Cells, 293T,Cells, HEK 293,Cells, HEK293,HEK 293 Cell,HEK293 Cell
D058504 Th17 Cells A subset of helper-effector T-lymphocytes which synthesize and secrete INTERLEUKINS IL-17; IL-17F; and IL-22. These cytokines are involved in host defenses and tissue inflammation in autoimmune diseases. T Helper 17 Cell,TH-17 Cell,Th17 Cell,Type 17 Helper T Cell,T Helper 17 Cells,TH-17 Cells,Type 17 Helper T Cells,Cell, TH-17,Cell, Th17,Cells, TH-17,Cells, Th17,TH 17 Cell,TH 17 Cells

Related Publications

Juan Liu, and Li-Ping Bai, and Fen Yang, and Xiaojun Yao, and Kawai Lei, and Christopher Wai Kei Lam, and Qibiao Wu, and Yuxin Zhuang, and Riping Xiao, and Kangsheng Liao, and Hioha Kuok, and Ting Li, and Liang Liu
December 2021, Phytochemistry,
Juan Liu, and Li-Ping Bai, and Fen Yang, and Xiaojun Yao, and Kawai Lei, and Christopher Wai Kei Lam, and Qibiao Wu, and Yuxin Zhuang, and Riping Xiao, and Kangsheng Liao, and Hioha Kuok, and Ting Li, and Liang Liu
January 2014, Natural product research,
Juan Liu, and Li-Ping Bai, and Fen Yang, and Xiaojun Yao, and Kawai Lei, and Christopher Wai Kei Lam, and Qibiao Wu, and Yuxin Zhuang, and Riping Xiao, and Kangsheng Liao, and Hioha Kuok, and Ting Li, and Liang Liu
May 2010, Chemical & pharmaceutical bulletin,
Juan Liu, and Li-Ping Bai, and Fen Yang, and Xiaojun Yao, and Kawai Lei, and Christopher Wai Kei Lam, and Qibiao Wu, and Yuxin Zhuang, and Riping Xiao, and Kangsheng Liao, and Hioha Kuok, and Ting Li, and Liang Liu
April 2021, Bioorganic chemistry,
Juan Liu, and Li-Ping Bai, and Fen Yang, and Xiaojun Yao, and Kawai Lei, and Christopher Wai Kei Lam, and Qibiao Wu, and Yuxin Zhuang, and Riping Xiao, and Kangsheng Liao, and Hioha Kuok, and Ting Li, and Liang Liu
July 2020, Journal of natural products,
Juan Liu, and Li-Ping Bai, and Fen Yang, and Xiaojun Yao, and Kawai Lei, and Christopher Wai Kei Lam, and Qibiao Wu, and Yuxin Zhuang, and Riping Xiao, and Kangsheng Liao, and Hioha Kuok, and Ting Li, and Liang Liu
October 2013, Steroids,
Juan Liu, and Li-Ping Bai, and Fen Yang, and Xiaojun Yao, and Kawai Lei, and Christopher Wai Kei Lam, and Qibiao Wu, and Yuxin Zhuang, and Riping Xiao, and Kangsheng Liao, and Hioha Kuok, and Ting Li, and Liang Liu
December 2016, Journal of ethnopharmacology,
Juan Liu, and Li-Ping Bai, and Fen Yang, and Xiaojun Yao, and Kawai Lei, and Christopher Wai Kei Lam, and Qibiao Wu, and Yuxin Zhuang, and Riping Xiao, and Kangsheng Liao, and Hioha Kuok, and Ting Li, and Liang Liu
August 2006, Journal of natural products,
Juan Liu, and Li-Ping Bai, and Fen Yang, and Xiaojun Yao, and Kawai Lei, and Christopher Wai Kei Lam, and Qibiao Wu, and Yuxin Zhuang, and Riping Xiao, and Kangsheng Liao, and Hioha Kuok, and Ting Li, and Liang Liu
February 2020, Journal of natural products,
Juan Liu, and Li-Ping Bai, and Fen Yang, and Xiaojun Yao, and Kawai Lei, and Christopher Wai Kei Lam, and Qibiao Wu, and Yuxin Zhuang, and Riping Xiao, and Kangsheng Liao, and Hioha Kuok, and Ting Li, and Liang Liu
October 2015, Bioorganic & medicinal chemistry letters,
Copied contents to your clipboard!