Abstract
Hedyotis hedyotidea has been used in traditional Chinese medicine for the treatment of autoimmune diseases. However, the mechanisms underlying for the effect remain unknown. We previously showed that, among 11 compounds extracted from H hedyotidea, betulin produced the strongest suppressive effect on T cell activation. Here, we examined the hepatoprotective effects of betulin against acute autoimmune hepatitis in mice and the mechanisms underlying the effects. Freshly isolated mouse splenocytes were stimulated with concanavalin A (Con A, 5 μg/mL) in the presence of betulin, the cell proliferation was assessed with CSFE-dilution assay. Mice were injected with betulin (10, 20 mg·kg−1·d−1, ip) for 3 d. One hour after the last injection, the mice were injected with Con A (15 mg/kg, iv) to induce acute hepatitis. Blood samples and liver tissues were harvested at 10 h after Con A injection, and serum transaminase levels and liver histopathology were detected; serum levels of proinflammatory cytokines, hepatic T lymphocyte ratios, and functional statuses of conventional T and NKT cells were also analyzed. Betulin (16 and 32 μmol/L) dose-dependently suppressed the proliferation of Con A-stimulated mouse splenocytes in vitro. In Con A-challenged mice, preinjection with betulin (20 mg·kg−1·d−1) significantly decreased the levels of proinflammatory cytokines IFN-γ, TNF-α and IL-6, and ameliorated liver injury. Furthermore, pretreatment with betulin (20 mg·kg−1·d−1) significantly inhibited the Con A-induced activation of NKT and conventional T cells, and decreased production of proinflammatory cytokines IFN-γ, TNF-α and IL-6 in these two cell populations. Betulin has immunomodulatory effect on overly activated conventional T and NKT cells and exerts hepatoprotective action in mouse autoimmune hepatitis. The findings provide evidence for the use of H hedyotidea and its constituent betulin in the treatment of autoimmune diseases.
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Acknowledgements
The authors would like to thank the NIH tetramer core facility for the CD1d tetramers, Dr Qin YANG of the Institute of Pathology, Tongji Hospital, Huazhong University of Science and Technology, and Senior Engineer Yaqin Wang and Associate Prof Xia WEI of the Department of Histology and Embryology, Medical College of China Three Gorges University for their help and technical assistance in histological examination. This work was supported by the National Natural Science Foundation of China (No 31370885, 31000150, and 81302528) and the 973 Program from the Ministry of Science and Technology of China (No 2013CB530505).
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Supplementary Figure S1
Betulin had little toxic effects on either splenocytes or L02 cells at the tested concentrations. (JPG 377 kb)
Supplementary Figure S2
Betulin did not affect the potential cytokine production capacity of any T cell subset stimulated with PMA and ionomycin. (JPG 665 kb)
Supplementary Table S1
1H NMR (400 MHz) and 13C NMR (100 MHz) data for betulin (CDCl3, J in Hz) (DOC 44 kb)
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Zhou, Yq., Weng, Xf., Dou, R. et al. Betulin from Hedyotis hedyotidea ameliorates concanavalin A-induced and T cell-mediated autoimmune hepatitis in mice. Acta Pharmacol Sin 38, 201–210 (2017). https://doi.org/10.1038/aps.2016.102
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DOI: https://doi.org/10.1038/aps.2016.102
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