Anti-sepsis protection of Xuebijing injection is mediated by differential regulation of pro- and anti-inflammatory Th17 and T regulatory cells in a murine model of polymicrobial sepsis☆
Introduction
Sepsis can cause mortality in > 40% of patients when systematic inflammation is out of control (Fink and Warren, 2014). The standard sepsis and septic shock management includes securing the airway, correcting hypoxemia, antibiotics and IV fluid (crystalloids and colloids), nutritional support and Corticosteroids (Dellinger et al., 2013, Sessler et al., 2004) (http://www.uptodate.com).
Cecal ligation and puncture (CLP), the most commonly used pre-clinical model of sepsis (Hubbard et al., 2005, Rittirsch et al., 2009), reflects clinical reality of polymicrobial sepsis (Anaya and Nathens, 2003). Multi-organ dysfunctional syndrome (MODS), endothelial dysfunction and depletion/inactivation of lymphocytes may contribute to the acute death in this model (Coletta et al., 2014, Hutchins et al., 2014). Serum level of IL-6 is a biomarker for survival and disease status in the CLP model (Iskander et al., 2013).
CD4+ T helper 17 cells (Th17) and Tregs play important roles in inflammatory diseases, such as sepsis and rheumatoid arthritis (Noack and Miossec, 2014). Th17 cells represent a pro-inflammatory subset whereas Treg cells promote an anti-inflammatory effect. As important immune regulators, Tregs are characterized by expressing transcription factor Foxp3, which plays a pivotal role in their development, lineage commitment, and regulatory functions (Fontenot et al., 2003, Hori et al., 2003).
Xuebijing injection (XBJ), a Chinese medicine formula combining 5 herbs, is routinely used as an add-on to conventional therapy to treat sepsis and septic shock in China (Jiang et al., 2013, Shi et al., 2016). Combination of XBJ and standard treatment yielded better clinical outcome than standard treatment alone (Gao et al., 2015, Shao et al., 2011). It reduces serum TNF-α concentrations in patients with multiple organ dysfunction syndrome (MODS) (Fang and Wang, 2013), but the mechanism for its attenuation of sepsis remains unclarified.
This study explored the regulatory effects of XBJ on CD4+ T cell differentiation and inflammation in a murine model of septic shock.
Section snippets
Chemicals and reagents
Xuebijing injection (catalogue number: z20040033, batch number: 1303261) was supplied by Tianjin Chase Sun Pharmaceutical Co., LTD (Tianjin, China). This Chinese medicine is approved by CFDA (China Food and Drug Administration) for treating sepsis and septic shock with CFDA ratification number of GuoYaoZhunZi-Z20040033 for market approval as a drug product. It is routinely used as an add-on to conventional therapy to treat sepsis and septic shock in China (Jiang et al., 2013). This injection
XBJ facilitates Treg expansion in CLP model and promotes Treg differentiation
There was a 10% increase of Treg upon 18 ml/kg XBJ treatment compared with CLP mice. While DEX slightly decreased Treg population in the spleen, frequencies of IL-10-positive splenocytes and Treg increased 4 fold upon 18 ml/kg XBJ treatment. In contrast, DEX and 6 ml/kg XBJ increased frequencies of IL-10-positive cells 2 fold in splenocytes and Tregs, indicating that 18 ml/kg XBJ might promote the expansion of functional Tregs (Fig. 1A-D). Thus, in vitro T cell differentiation assay was conducted
Discussion
The goal of this study is to access the immuno-regulatory role of Chinese herbal medicine XBJ in a septic shock model. We discovered that 18 ml/kg XBJ not only increased the frequency of Tregs in septic mice but also induced Treg differentiation in vitro. In contrast, it normalized pro-inflammatory Th17 population in septic mice and inhibited Th17 differentiation in vitro. Accordingly, 18 ml/kg XBJ significantly reduced systemic pro-inflammatory cytokines, IL-6 and TNF-α, in serum and reduced
Acknowledgments
We would like to thank the members of our laboratory, particularly Dr. Zhaochen Ning, Ming Lv, Jing Han, Qunqun Du for sharing reagents, providing constructive discussions and ideas.
Author contributions
XC and YZ conceptualized the ideas of this work. XS and XC carried out all the experiments. XC, XS, YF and YZ wrote the original draft paper. XC, YF, GP, GF and YZ analyzed the experimental results and revised the paper. XG JH supervised the work and contributed to the study conceptualization.
Conflict of interest
The authors declare no conflict of interest.
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Cited by (0)
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This study was supported by grants from the National Key Basic Research Program of China (2012CB723504), the National Major New Drug Discovery (2013ZX0920102), and the National Science Foundation of China (NSFC 81274128).
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Equal contribution to this work.