Coptidis alkaloids extracted from Coptis chinensis Franch attenuate IFN-γ-induced destruction of bone marrow cells.

Jinyu Li,Huijie Zhang,Xiaoying Meng,Jie Wei,Changzhi Wang,Meiyier Huandike,Hening Chen,Limin Chai,Peiying Deng,Juan Liu,Xing-Ming Shi

doi:10.1371/journal.pone.0236433

Abstract

Coptidis alkaloids are the primary active components of Coptis chinensis Franch. Clinical and pharmacodynamic studies have confirmed that Coptidis alkaloids have multiple therapeutic effects including anti-inflammatory, antioxidant and antitumor effects, and they are usually used to treat various inflammatory disorders and related diseases. Mouse bone marrow cells (BMCs) were isolated from BALB/c mice. Immune-mediated destruction of BMCs was induced by interferon (IFN) -γ. High-performance liquid chromatography-electrospray ionization/ mass spectrometry was used to analyze the ingredients of the aqueous extract from Coptis chinensis Franch. The results confirmed that Coptidis alkaloids were the predominant ingredients in the aqueous extract from Coptis chinensis. The functional mechanism of Coptidis alkaloids in inhibiting immune-mediated destruction of BMCs was studied in vitro. After Coptidis alkaloid treatment, the percentages of apoptotic BMCs and the proliferation and differentiation of helper T (Th) cells and regulatory T (Treg) cells were measured by flow cytometry. The expression and distribution of T-bet in BMCs were observed by immunofluorescence. Western blotting analysis was used to assay the expression of key molecules in the Fas apoptosis and Jak/Stats signaling pathways in BMCs. We identified five alkaloids in the aqueous extract of Coptis chinensis. The apoptotic ratios of BMCs induced by IFN-γ were decreased significantly after Coptidis alkaloid treatment. The levels of key molecules (Fas, Caspase-3, cleaved Caspase-3, Caspase-8 and Caspase-8) in Fas apoptosis signaling pathways also decreased significantly after treatment with low concentrations of Coptidis alkaloids. Coptidis alkaloids were also found to inhibit the proliferation of Th1 and Th17 cells and induce the differentiation of Th2 and Treg cells; further, the distribution of T-bet in BMCs was decreased significantly. In addition, the levels of Stat-1, phospho-Stat-1 and phospho-Stat-3 were also reduced after Coptidis alkaloid treatment. These results indicate that Coptidis alkaloids extracted by water decoction from Coptis chinensis Franch could inhibit the proliferation and differentiation of T lymphocytes, attenuate the apoptosis of BMCs, and suppress the immune-mediated destruction of the BMCs induced by pro-inflammatory cytokines.

Highlights

The activation and expansion of self-reactive pathogenic T cells and highly secreted inflammatory cytokines, such as interferon gamma (IFN-γ) and tumor necrosis factor α (TNFα), can impair the signaling of several cytokine receptors, mediate inflammation and target cell destruction
The levels of Stat-1, phospho-Stat-1 and phospho-Stat-3 were reduced after Coptidis alkaloid treatment. These results indicate that Coptidis alkaloids extracted by water decoction from Coptis chinensis Franch could inhibit the proliferation and differentiation of T lymphocytes, attenuate the apoptosis of bone marrow cells (BMCs), and suppress the immunemediated destruction of the BMCs induced by pro-inflammatory cytokines
There were small amounts of other alkaloids in the aqueous extract, including magnoflorine, jatrorrhizine, epiberberine and coptisineis. These findings suggested that the Coptidis alkaloids extracted from Coptis chinensis by water decoction had regulatory effects on the inflammatory response and cell apoptosis of BMCs induced by pro-inflammatory cytokines

Summary

Introduction

The activation and expansion of self-reactive pathogenic T cells and highly secreted inflammatory cytokines, such as interferon gamma (IFN-γ) and tumor necrosis factor α (TNFα), can impair the signaling of several cytokine receptors, mediate inflammation and target cell destruction. These pathological changes are the key cellular events in the development of immune-mediated bone marrow (BM) failure. Helper T (Th) 1 cells and their response contribute to these negative effects [1,2,3] Both the Fas/FasL and perforin/granzyme pathways are involved in cytotoxic T cell-induced targeting of cells for apoptosis, resulting in the immunemediated BM destruction [4]. The protein levels of Fas, caspases and other proapoptotic genes was shown to significantly increased in BM cells induced by IFN-γ [1]

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