Role of TOLL-like receptor signal transduction pathway in inflammatory reaction induced by endoplasmic reticulum stress

doi:10.3877/cma.j.issn.1674-1358.2017.03.004

Abstract

Abstract:

Objective

To investigate the role of Toll-like receptor signal transduction pathway on the process of endoplasmic reticulum stress and inflammatory reaction induced by tunicamycin (TM).

Methods

The transcription levels of IRAK1, TAK1 and NF-κB genes in HepG2 and L02 cells treated by TM were detected by real-time reverse transcription PCR, and the expression level of protein related cytokines was detected by Western blot.

Results

The levels of GRP78 (marker of endoplasmic reticulum stress) in HepG2 cells and L02 cells significantly increased after treated by TM. The gene transcription and protein expression level of NF-κB (marker of inflammatory response) were both significantly increased in accordance with the concentration and action time compared with those untreated by TM. In Toll-like receptor signal transduction pathway, the expression of key cytokines like IRAK1 and TAK1 showed an increasing trend at gene and protein levels, all with significant differences (all P < 0.05).

Conclusions

After the stimulation of TM, endoplasmic reticulum stress and inflammatory reaction were induced in HepG2 and L02 cell lines. In the endoplasmic reticulum stress induced activation inflammatory reaction associated with Toll-like receptor signaling pathway, Toll-like receptor signal transduction pathway in the occurrence and development may have a regulatory role.

Key words: Endoplasmic reticulum stress, Inflammatory reaction, Toll-like receptor signal transduction pathway, Tunicamycin

Siyuan Yang, Hui Li, Xin Li, Rongmeng Jiang, Chengjie Ma, Hongshan Wei, Xingwang Li. Role of TOLL-like receptor signal transduction pathway in inflammatory reaction induced by endoplasmic reticulum stress[J]. Chinese Journal of Experimental and Clinical Infectious Diseases(Electronic Edition), 2017, 11(03): 222-227.

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URL: https://zhsyhlcgrbzz.cma-cmc.com.cn/EN/10.3877/cma.j.issn.1674-1358.2017.03.004

https://zhsyhlcgrbzz.cma-cmc.com.cn/EN/Y2017/V11/I03/222

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References 26

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引物名称	引物序列（5´→3´）
β-actin	F：attggcaatgagcggttc
β-actin	R：cgtggatgccacaggact
IRAK1	F：tgccaggaccaagtatctga
IRAK1	R：gtgcttctcaaagccactcc
TAK1	F：agctttatggagcctgcttg
TAK1	R：agagcccccttcagcatatt
NF-κB	F：atgcgcttccgctacaagt
NF-κB	R：tggtggtatctgtgctccttc

引物名称	引物序列（5´→3´）
β-actin	F：attggcaatgagcggttc
β-actin	R：cgtggatgccacaggact
IRAK1	F：tgccaggaccaagtatctga
IRAK1	R：gtgcttctcaaagccactcc
TAK1	F：agctttatggagcctgcttg
TAK1	R：agagcccccttcagcatatt
NF-κB	F：atgcgcttccgctacaagt
NF-κB	R：tggtggtatctgtgctccttc

组别	目的基因相对表达量
	IRAK1		TAK1		NF-Κb
	HepG2	L02	HepG2	L02	HepG2	L02
空白对照组	0.111 ± 0.113	0.130 ± 0.027	0.197 ± 0.007	0.213 ± 0.036	0.129 ± 0.028	0.090 ± 0.013
5 mg/ml刺激12 h组	0.527 ± 0.190	0.346 ± 0.101	0.412 ± 0.061	0.452 ± 0.100	0.401 ± 0.032	0.415 ± 0.059
5 mg/ml刺激24 h组	1.106 ± 0.215	0.903 ± 0.081	0.894 ± 0.081	0.824 ± 0.052	0.766 ± 0.048	0.756 ± 0.060
10 mg/ml刺激24 h组	1.263 ± 0.249	1.073 ± 0.189	0.983 ± 0.115	0.994 ± 0.112	1.007 ± 0.122	0.920 ± 0.029
P₁	0.003	0.010	0.068	0.545	0.631	0.051
P₂	0.015	0.002	0.016	0.020	0.033	0.018
P₃	< 0.001	< 0.001	< 0.001	< 0.001	< 0.001	< 0.001

组别	目的基因相对表达量
	IRAK1		TAK1		NF-Κb
	HepG2	L02	HepG2	L02	HepG2	L02
空白对照组	0.111 ± 0.113	0.130 ± 0.027	0.197 ± 0.007	0.213 ± 0.036	0.129 ± 0.028	0.090 ± 0.013
5 mg/ml刺激12 h组	0.527 ± 0.190	0.346 ± 0.101	0.412 ± 0.061	0.452 ± 0.100	0.401 ± 0.032	0.415 ± 0.059
5 mg/ml刺激24 h组	1.106 ± 0.215	0.903 ± 0.081	0.894 ± 0.081	0.824 ± 0.052	0.766 ± 0.048	0.756 ± 0.060
10 mg/ml刺激24 h组	1.263 ± 0.249	1.073 ± 0.189	0.983 ± 0.115	0.994 ± 0.112	1.007 ± 0.122	0.920 ± 0.029
P₁	0.003	0.010	0.068	0.545	0.631	0.051
P₂	0.015	0.002	0.016	0.020	0.033	0.018
P₃	< 0.001	< 0.001	< 0.001	< 0.001	< 0.001	< 0.001

组别	灰度分值比较
组别	IRAK1	TAK1	pp65	GRP78
空白对照组	1.000	1.000	1.000	1.000
5 mg/ml 12 h组	1.033 ± 0.023	1.432 ± 0.372	1.020 ± 0.012	1.038 ± 0.033
5 mg/ml 24 h组	1.925 ± 0.073	2.095 ± 0.481	1.236 ± 0.048	1.326 ± 0.271
10 mg/ml 24 h组	2.016 ± 0.049	2.882 ± 0.715	1.507 ± 0.092	1.926 ± 0.249
P₁值	0.007	0.015	0.399	0.001
P₂值	< 0.001	0.010	0.001	0.005
P₃值	< 0.001	0.019	< 0.001	< 0.001

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