大鼠骨髓树突状细胞的分离培养及HMGB1/TLR4信号通路的调控作用

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Abstract Objective: To investigate the regulatory effect of HMGB1/TLR4 signaling pathway on the expression of MyD88, NF-κB P65, cytokine and costimulatory molecules in rat bone marrow-derived dendritic cells (mDCs). Methods: Dendritic cells were isolated, cultured, purified and identified in vitro from bone marrow stem cells of rats, then randomly divided into 5 groups: mDCs cultured with HMGB1 only (H group), HMGB1 specific neutralizing antibody (H-Ab group), TLR4 antagonist (TLR4-A group) and control IgG (IgG group) 30 min after HMGB1, separately, or with RPMI l640 serum-containing medium as control group (C gruop). Each group was observed at different time points of 24 h, 48 h and 72 h by detecting the expression of MyD88, NF-κB P65 protein and mRNA, costimulatory molecules (CD80, CD86) and the levels of cytokine (IL-6, IL-8, IL-12, TNF-α) in the supernatant. Results: MyD88 and NF-κB P65 protein and mRNA, costimulatory molecules and cytokine levels in mDCs stimulated with HMGB1 were significantly higher than control group at each time point (P<0.01). The levels were significantly decreased after being treated with HMGB1 specific neutralizing antibody or TLR4 antagonist (P<0.01). Conclusion: Data analysis suggests that HMGB1 plays a regulatory role in mDCs by affecting the downstream MyD88, NF-κB P65 protein and mRNA expression through TLR4, stimulating the secretion of cytokine and promoting the expression of costimulatory molecules (CD80, CD86).

Key words： high mobility group box 1 protein Toll-like receptor-4 signaling pathway dendritic cells rats

Received: 26 November 2017

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	WANG Hanlei
	XUE Jiyang
	GE Hanwei
	XIA Jie
	LIN Wei
	ZHAO Qifeng.

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https://xb.wmu.edu.cn/EN/ OR https://xb.wmu.edu.cn/EN/Y2018/V48/I7/508

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