The Immuno-Enhancement Effects of Tubiechong (Eupolyphaga sinensis) Lyophilized Powder in Cyclophosphamide-Induced Immunosuppressed Mice

ABSTRACT

Tubiechong (Eupolyphaga sinensis) is an important material used in traditional Chinese medicine (TCM). However, the immunoregulation effects of E. sinensis Lyophilized Powder (ESL) are unclear. The in vivo study thus designed to elucidate the immuno-enhancement effects of ESL in immunosuppressed mice induced by cyclophosphamide (CTX). Mice were treated with three doses of ESL (0.5, 1.0 and 2.0 g/kg). Compared with model group, ESL notably increased the immune organ index, mononuclear macrophages function and the level of nature killer cell (NK) (p < 0.05 or p < 0.01), delayed type hypersensitivity (DTH) was also improved (p < 0.05). The level of superoxide dismutase (SOD) and catalase (CAT) were enhanced (p < 0.05), while malonyldialdehyde (MDA) and nitrogen monoxide (NO) were reduced (p < 0.05 or p < 0.01). Meanwhile, cluster determinant (CD)3⁺ T cell, CD4⁺ T cell and CD4⁺/CD8⁺ ratio were increased (p < 0.01). The cytokines secretion such as interleukin (IL)-2 and tumor necrosis factor alpha (TNF-α) were notably increased (p < 0.05 or p < 0.01), and IL-6 and IL-16 were also enhanced (p < 0.05). Furthermore, ESL significantly inhibited the phosphorylation of c-Jun N-terminal kinase (JNK), down-regulated the expression of Bcl-2 associated X protein (Bax), up-regulated the B cell lymphoma-2 protein (Bcl-2) expression and decreased the Bax/Bcl-2 ratio in spleen tissues (p < 0.05). In brief, all these findings suggest that ESL could effectively improve immune functions via modulating oxidative systems and innate immune cells.

Abbreviations: TCM: Traditional Chinese Medicine; ESL: Eupolyphaga sinensis Lyophilized Powder; CCl₄: Carbon tetrachloride; ERK: Extracellular regulated protein kinases; CTX: Cyclophosphamide; DTH: Delayed type hypersensitivity; SOD: Superoxide dismutase; CAT: Catalase; MDA: Malonyldialdehyde; NO: Nitrogen monoxide; NK: Nature killer cell; CD: Cluster determinant interleukin; TNF-α: Tumor Necrosis Factor alpha; JNK: c-Jun N-terminal kinase; Bax: Bcl-2 associated X protein; Bcl-2: B cell lymphoma-2 protein; Th1: Type-1 helper; Th2: Type-2 helper; FAMEs: Fatty acid methyl esters; DNFB: 2,4 – Dinitrofluorobenzene; ELISA: Enzyme-linked immuno sorbent assay; MAPK: Mitogen activated protein kinase; Cyt-c: Cytochrome c; SCFAs: Short-chain fatty acids; SDS-PAGE: Sodium dodecyl sulfate polyacrylamide gel electrophoresis

Graphical Abstract

KEYWORDS:

• Eupolyphaga sinensis Lyophilized Powder
• immuno-enhancement
• anti-oxidation
• immune cells

Authors’ Contributions

Huiyun Liu performed the experiments and wrote the manuscript; Yunliang Yan performed the data analysis and risk assessment; Fengling Zhang conceived the project and polished the writing; Qiaofeng Wu conceived the project, protocol development, developed the research strategy and wrote the manuscript.

Acknowledgments

This work was supported by the Science Technology Department of Zhejiang Province (2011C22004); National Natural Science Foundation of China (81274030/H2803, 81473335/H2803); Science Foundation of Zhejiang Chinese Medical University (2015ZR07) and Zhejiang Provincial Natural Science Foundation of China (LY15H290003/LY18H280007).

Conflicts of Interest

The authors declare that there is no conflict of interest regarding the publication of this paper.

Additional information

Funding

This work was supported by the National Natural Science Foundation of China [81274030/H2803, 81473335/H2803].