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Published on: April 3, 2017
Yak-Derived CXCL14 Activates the Pro-Inflammatory Response of Macrophages and Inhibits the Proliferation and
Biao Li1,2, Juan Li1,2, Li Wang1,2
1Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization, Ministry of Education, Southwest Minzu University, Chengdu 610041, China.
Abstract:
CXCL14 (C-X-C motif chemokine ligand 14) is an important chemokine involved in infection and immunity and plays an important role in a variety of immune-related diseases. The 446 bp cDNA sequence of the CXCL14 gene in yaks was obtained. Additionally, the prokaryotic expression vector of the CXCL14 protein with a molecular weight of 27 kDa was successfully constructed and expressed. The proliferation activities and migration abilities of spleen macrophages were significantly inhibited after treatment with the CXCL14 protein at different concentrations (1, 10 and 20 μg/mL) (p < 0.05). Furthermore, the expressions of pro-inflammatory cytokines interleukin 1 beta (IL-1β), interleukin 6 (IL6), interleukin 8 (IL8) and interferon-α (TNF-α) were significantly increased (p < 0.05), but the expression of anti-inflammatory factor interleukin 10 (IL10) was significantly decreased (p < 0.05). The contents of inflammatory factors in the supernatant of cells were detected using ELISA, and it was also found that the contents of TNF-α, IL6 and cytochrome c oxidase subunit II (COX2) were significantly increased under different CXCL14 protein concentrations (p < 0.05). Finally, the exogenous addition of CXCL14 inhibited the activity, clonal formation and migration of hepatoma cells (HepG2). Additionally, after HepG2 cells were treated with 20 μg/mL CXCL14 protein for 12 h, 24 h and 36 h, the expression levels of BCL2 homologous antagonist/killer (BAK) and the BCL2-associated X apoptosis regulator (BAX) were increased to varying degrees, while the expression levels of hypoxia-inducible factor 1 subunit alpha (HIF1A), the mechanistic target of rapamycin kinase (mTOR) and cyclin-dependent kinase 1 (CDK1) genes decreased compared to the control group. In conclusion, the CXCL14 protein can inhibit the proliferation and migration of HepG2 cells by inducing the expression of macrophage pro-inflammatory factors and activating apoptosis-related genes to exert innate immunity. These results are helpful to further study the function of the CXCL14 protein and provide research data for the innate immune mechanism of yaks under harsh plateau environments.
Insights
Yak CXCL14 protein inhibits spleen macrophage activity and hepatoma cell growth by modulating inflammatory responses and apoptosis. This research provides insights into innate immunity in yaks facing harsh plateau environments.
Area of Science:
- Immunology
- Molecular Biology
- Yak Research
Background:
- Chemokines like CXCL14 (C-X-C motif chemokine ligand 14) are crucial in immune responses and diseases.
- Understanding yak immune mechanisms is vital for adapting to harsh plateau environments.
Purpose of the Study:
- To investigate the function of yak CXCL14 protein in immune regulation and its effect on hepatoma cells.
- To explore the role of CXCL14 in innate immunity.
Main Methods:
- Obtained yak CXCL14 cDNA sequence and constructed a prokaryotic expression vector.
- Treated spleen macrophages and HepG2 hepatoma cells with purified CXCL14 protein.
- Analyzed cell proliferation, migration, cytokine expression (IL-1β, IL6, IL8, TNF-α, IL10), inflammatory markers (COX2), and apoptosis-related genes (BAK, BAX, HIF1A, mTOR, CDK1).
Main Results:
- Yak CXCL14 protein significantly inhibited spleen macrophage proliferation and migration.
- CXCL14 treatment increased pro-inflammatory cytokines (TNF-α, IL6) and decreased anti-inflammatory IL10 in macrophages.
- CXCL14 inhibited HepG2 cell activity, clonal formation, and migration, while upregulating apoptosis genes (BAK, BAX) and downregulating proliferation-related genes (HIF1A, mTOR, CDK1).
Conclusions:
- Yak CXCL14 protein exerts innate immunity by inhibiting macrophage pro-inflammatory responses and inducing apoptosis in hepatoma cells.
- These findings contribute to understanding yak immune function in challenging environments and offer potential therapeutic targets.
