Novel roles of lipopolysaccharide and TLR4/NF-κB signaling pathway in inflammatory response to liver injury in Budd-Chiari syndrome

doi:10.4240/wjgs.v13.i11.1448

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World Journal of Gastrointestinal Surgery

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1948-9366

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Baishideng Publishing Group Inc, 7041 Koll Center Parkway, Suite 160, Pleasanton, CA 94566, USA

Observational Study

World J Gastrointest Surg. Nov 27, 2021; 13(11): 1448-1462
Published online Nov 27, 2021. doi: 10.4240/wjgs.v13.i11.1448

Table 1 Primers used in this study

	Forward	Reverse
β-actin (150 bp)	5’-CCCATCTATGAGGGTTACGC-3’	5’-TTTAATGTCACGCACGATTTC-3’
TLR4 (186 bp)	5’-GCCGGAAAGTTATTGTGGTGGT-3’	5’-ATGGGTTTTAGGCGCAGAGTTT-3’
NF-κB p65 (108 bp)	5’-AAGATCTGCCGAGTAAACCG-3	5’-TCCCGTGAAATACACCTCAA-3’
IL-2 (113 bp)	5’-CAAGCAGGCCACAGAATTGA-3’	5’-TTCCAGCGTCTTCCAAGTGA-3’
TNF-α (89 bp)	5’-AGGAGGGAGAACAGCAACTC-3’	5’-TGTATGAGAGGGACGGAACC-3’
IFN-γ (130 bp)	5’-CAGGCCATCAGCAACAACAT-3’	5’-GCTGGATCTGTGGGTTGTTC-3’

TLR4: Toll-like receptor 4; NF-κB: Nuclear factor-κB; TNF-α: Tumor necrosis factor-α; IL-2: Interleukin-2; IFN-γ: Interferon-γ.

Table 2 mRNA expression levels of TLR4/NF-κB-mediated inflammation-related indicators

	TLR4	NF-κBp65	IL-2	TNF-α	IFN-γ
Control group	1.004 ± 0.139	1.003 ± 0.074	1.001 ± 0.121	1.001 ± 0.126	1.005 ± 0.101
Sham-operated group
1W	1.101 ± 0.127	1.101 ± 0.127	1.108 ± 0.206	1.068 ± 0.222	1.102 ± 0.121
3W	1.013 ± 0.109	1.013 ± 0.109	1.061 ± 0.168	1.042 ± 0.181	1.082 ± 0.111
6W	1.045 ± 0.118	1.045 ± 0.118	1.006 ± 0.141	1.047 ± 0.164	1.004 ± 0.084
12W	1.059 ± 0.115	1.059 ± 0.115	1.025 ± 0.097	1.017 ± 0.157	1.019 ± 0.118
Model group
1W	1.773 ± 0.165^a^,^b	1.773 ± 0.165^a^,^b	1.935 ± 0.217^a^,^b	1.991 ± 0.181^a^,^b	2.170 ± 0.195^a^,^b
3W	6.789 ± 0.692^a^,^b	6.789 ± 0.692^a^,^b	6.144 ± 0.681^a^,^b	12.931 ± 1.424^a^,^b	9.172 ± 0.902^a^,^b
6W	4.671 ± 0.593^a^,^b	4.671 ± 0.593^a^,^b	4.372 ± 0.268^a^,^b	7.629 ± 0.999^a^,^b	5.131 ± 0.441^a^,^b
12W	1.003 ± 0.074^a^,^b	3.575 ± 0.334^a^,^b	3.011 ± 0.326^a^,^b	3.991 ± 0.391^a^,^b	2.855 ± 0.732^a^,^b
Statistics	333.288	464.025	426.396	555.318	509.268
P value	< 0.0001	< 0.0001	< 0.0001	< 0.0001	< 0.0001

^aP < 0.05, comparisons between subgroups, the differences are statistically significant.

^bP < 0.05, comparisons between model subgroups, the differences are statistically significant.

TLR4: Toll-like receptor 4; NF-κB: Nuclear factor-κB; TNF-α: Tumor necrosis factor-α; IL-2: Interleukin-2; IFN-γ: Interferon-γ.

Table 3 Relative protein expression levels of TLR4/NF-κB-mediated inflammation-related indicators

	TLR4	NF-κBp65	IL-2	TNF-α	IFN-γ
Control group	0.555 ± 0.066	0.317 ± 0.022	0.232 ± 0.029	0.368 ± 0.032	0.369 ± 0.023
Sham-operated group
1W	0.561 ± 0.059	0.327 ± 0.019	0.229 ± 0.028	0.366 ± 0.026	0.379 ± 0.029
3W	0.560 ± 0.068	0.313 ± 0.023	0.242 ± 0.029	0.357 ± 0.022	0.359 ± 0.026
6W	0.561 ± 0.071	0.310 ± 0.030	0.236 ± 0.027	0.369 ± 0.034	0.362 ± 0.020
12W	0.564 ± 0.070	0.309 ± 0.025	0.234 ± 0.027	0.371 ± 0.028	0.377 ± 0.028
Model group
1W	0.729 ± 0.067^a^,^b	0.678 ± 0.058^a^,^b	0.449 ± 0.057^a^,^b	0.684 ± 0.024^a^,^b	0.700 ± 0.082^a^,^b
3W	1.507 ± 0.086^a^,^b	1.416 ± 0.087^a^,^b	1.047 ± 0.058^a^,^b	1.188± 0.051^a^,^b	1.261 ± 0.073^a^,^b
6W	1.162 ± 0.070^a^,^b	1.192 ± 0.079^a^,^b	0.848 ± 0.056^a^,^b	1.050 ± 0.043^a^,^b	1.069 ± 0.090^a^,^b
12W	1.001 ± 0.072^a^,^b	0.880 ± 0.090^a^,^b	0.610 ± 0.059^a^,^b	0.835 ± 0.065^a^,^b	0.845 ± 0.063^a^,^b
Statistics	291.836	711.802	608.214	897.062	488.525
P value	< 0.0001	< 0.0001	< 0.0001	< 0.0001	< 0.0001

^aP < 0.05, comparisons between subgroups, the differences are statistically significant.

^bP < 0.05, comparisons between model subgroups, the differences are statistically significant.

TLR4: Toll-like receptor 4; NF-κB: Nuclear factor-κB; TNF-α: Tumor necrosis factor-α; IL-2: Interleukin-2; IFN-γ: Interferon-γ.

Table 4 Correlation between lipopolysaccharide and mRNA levels of various inflammation-related indicators

	LPS	TLR4	NF-κBp65	IFN-γ	TNF-α	IL-2
LPS	1	0.959^a	0.945^a	0.939^a	0.942^a	0.944^a
NF-κBp65	0.945^a	0.930^a	1	0.941^a	0.954^a	0.966^a
IL-2	0.939^a	0.952^a	0.941^a	1	0.982^a	0.946^a
TNF-α	0.942^a	0.953^a	0.954^a	0.982^a	1	0.954^a
IFN-γ	0.944^a	0.932^a	0.966^a	0.946^a	0.954^a	1

^aP < 0.01.

LPS: Lipopolysaccharide; TLR4: Toll-like receptor 4; NF-κB: Nuclear factor-κB; TNF-α: Tumor necrosis factor-α; IL-2: Interleukin-2; IFN-γ: Interferon-γ.

Table 5 Correlation between lipopolysaccharide and protein expression of inflammation-related indicators

	LPS	TLR4	NF-κBp65	IFN-γ	TNF-α	IL-2
LPS	1	0.954^a	0.929^a	0.931^a	0.930^a	0.936^a
TLR4	0.954^a	1	0.915^a	0.900^a	0.930^a	0.910^a
NF-κBp65	0.929^a	0.915^a	1	0.954^a	0.949^a	0.961^a
IFN-γ	0.931^a	0.900^a	0.954^a	1	0.938^a	0.957^a
TNF-α	0.930^a	0.930^a	0.949^a	0.938^a	1	0.954^a
IL-2	0.936^a	0.910^a	0.961^a	0.957^a	0.954^a	1

^aP < 0.01.

LPS: Lipopolysaccharide; TLR4: Toll-like receptor 4; NF-κB: Nuclear factor-κB; TNF-α: Tumor necrosis factor-α; IL-2: Interleukin-2; IFN-γ: Interferon-γ.

Table 6 Levels of lipopolysaccharide and TLR4/NF-κB-mediated inflammation-related indicators in the blood samples from human subjects

	LPS (Eu/mL)	TLR4 (%)	NF-κB (pg/mL)	IL-2 (pg/mL)	TNF-α (pg/mL)	IFN-γ (pg/mL)
Control group	8.42 ± 2.33	5.05 ± 1.29	8.15 ± 1.65	21.19 ± 5.01	24.88 ± 4.07	16.60 ± 1.80
Chronic group	20.96 ± 6.70	14.00 ± 3.67	28.75 ± 4.17	65.62 ± 13.26	46.68 ± 7.55	35.87 ± 7.36
Acute group	34.44 ± 10.45^a^,^b	24.55 ± 7.0^a^,^b	65.17 ± 11.09^a^,^b	127.90 ± 26.57^a^,^b	77.88 ± 9.61^a^,^b	70.90 ± 11.95^a^,^b
F	95.541	132.171	524.000	285.085	384.673	340.340

^aP < 0.05, Acute group vs Chronic group, the differences are statistically significant.

^bP < 0.05, Acute group vs Control group, the differences are statistically significant.

LPS: Lipopolysaccharide; TLR4: Toll-like receptor 4; NF-κB: Nuclear factor-κB; TNF-α: Tumor necrosis factor-α; IL-2: Interleukin-2; IFN-γ: Interferon-γ.

Citation: Li J, Chen XM, Zhou CZ, Fang WW, Lv WF, Cheng DL. Novel roles of lipopolysaccharide and TLR4/NF-κB signaling pathway in inflammatory response to liver injury in Budd-Chiari syndrome. World J Gastrointest Surg 2021; 13(11): 1448-1462
URL: https://www.wjgnet.com/1948-9366/full/v13/i11/1448.htm
DOI: https://dx.doi.org/10.4240/wjgs.v13.i11.1448