Effect of nucleotide binding oligomerization domain-like receptor protein 3 inflammasome on kidney tissues in septic rats with acute kidney injury

doi:10.3877/cma.j.issn.1674-6880.2020.01.011

Abstract

Abstract:

Objective

To investigate the role and mechanism of nucleotide binding oligomerization domain-like receptor protein 3 (NLRP3) inflammasome in septic rats with acute kidney injury (AKI).

Methods

A total of 18 male Wistar rats were randomly divided into a lipopolysaccharide (LPS) group, a LPS + inhibitor group, and a negative control group, 6 in each group. Rats in the LPS group were given an intraperitoneal injection of LPS (3.5 mg/kg), and rats in the LPS + inhibitor group were given an intraperitoneal injection of LPS (3.5 mg/kg) and cysteinyl aspartate specific proteinase-1 (Caspase-1) inhibitor Bernacasan (VX-765, 100 mg/kg), while rats in the negative control group were given an intraperitoneal injection of isotonic NaCl solution (0.3 mL). Serum creatinine levels of rats in the 3 groups were compared before and after modeling for 24 h. The expression levels of serum interleukin-1beta (IL-1β), IL-18, and tumor necrosis factor-alpha (TNF-α) of rats in the 3 groups were measured by enzyme-linked immunosorbent assay (ELISA) after modeling for 24 h. In the meantime, the expression levels of NLRP3 and Caspase-1 proteins were detected by Western-blotting after modeling for 24 h.

Results

The serum creatinine levels of rats in the 3 groups were statistically significantly different before and after modeling (F = 146.910, P < 0.001). Further pairwise comparisons showed that after modeling, the serum creatinine levels in the LPS + inhibitor group and negative control group were both significantly lower than those in the LPS group[(1.57 ± 0.20), (0.54 ± 0.39), (2.31 ± 0.24) mg/L], and the serum creatinine level in the negative control group [(0.54 ± 0.39) mg/L vs. (1.57 ± 0.20) mg/L] was lower than that in the LPS + inhibitor group, whereas the serum creatinine levels in the LPS group [(2.31 ± 0.24) mg/L vs. (0.53 ± 0.16) mg/L] and LPS + inhibitor group [(1.57 ± 0.20) mg/L vs. (0.56 ± 0.08) mg/L] after modeling were both significantly higher than those before modeling (all P < 0.05). The expression levels of serum IL-1β [(9.33 ± 1.16), (1.93 ± 0.30), (1.88 ± 0.30) ng/L], IL-18 [(23.8 ± 2.8), (19.6 ± 1.5), (16.6 ± 1.2) ng/L], TNF-α [(42.3 ± 2.1), (23.9 ± 2.5), (23.5 ± 1.3) ng/L], NLRP3 proteins [(2.04 ± 0.25), (2.04 ± 0.27), (1.49 ± 0.28)], and Caspase-1 proteins [(0.62 ± 0.07), (0.51 ± 0.09), (0.50 ± 0.09)] were statistically significantly different among the LPS group, LPS + inhibitor group and negative control group (F = 217.015, 20.590, 168.766, 8.723, 3.905; all P < 0.05). Further comparisons showed that the serum levels of IL-1β, IL-18, and TNF-α in the LPS + inhibitor group and negative control group were significantly lower than those in the LPS group, and the IL-18 level in the negative control was lower than that in the LPS + inhibitor group (all P < 0.05). The levels of NLRP3 protein in the negative control group were significantly lower than those both in the LPS group and the LPS + inhibitor group (both P < 0.05). In addition, the expression levels of Caspase-1 protein in the LPS + inhibitor group and negative control group were both significantly lower than those in the LPS group (both P < 0.05).

Conclusion

In septic rats, the NLRP3 inflammasome is activated and then acute kidney damage occurs through the Caspase-1 pathway.

Key words: Sepsis, Nucleotide binding oligomerization domain-like receptor protein 3, Acute kidney injury

Jiannan Zhang, Wen Liu, Guangping Chang, Yanhui Cao. Effect of nucleotide binding oligomerization domain-like receptor protein 3 inflammasome on kidney tissues in septic rats with acute kidney injury[J]. Chinese Journal of Critical Care Medicine(Electronic Edition), 2020, 13(01): 55-59.

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References 22

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组别	例数	建模前	建模后
LPS组	6	0.53 ± 0.16	2.31 ± 0.24^a
LPS +抑制剂组	6	0.56 ± 0.08	1.57 ± 0.20^ab
阴性对照组	6	0.53 ± 0.10	0.54 ± 0.39^bc
F值	?	146.910
P值	?	< 0.001

组别	例数	建模前	建模后
LPS组	6	0.53 ± 0.16	2.31 ± 0.24^a
LPS +抑制剂组	6	0.56 ± 0.08	1.57 ± 0.20^ab
阴性对照组	6	0.53 ± 0.10	0.54 ± 0.39^bc
F值	?	146.910
P值	?	< 0.001

组别	例数	IL-1β	IL-18	TNF-α
LPS组	6	9.33 ± 1.16	23.8 ± 2.8	42.3 ± 2.1
LPS +抑制剂组	6	1.93 ± 0.30^a	19.6 ± 1.5^a	23.9 ± 2.5^a
阴性对照组	6	1.88 ± 0.30^a	16.6 ± 1.2^ab	23.5 ± 1.3^a
F值	?	217.015	20.590	168.766
P值	?	< 0.001	< 0.001	< 0.001

组别	例数	IL-1β	IL-18	TNF-α
LPS组	6	9.33 ± 1.16	23.8 ± 2.8	42.3 ± 2.1
LPS +抑制剂组	6	1.93 ± 0.30^a	19.6 ± 1.5^a	23.9 ± 2.5^a
阴性对照组	6	1.88 ± 0.30^a	16.6 ± 1.2^ab	23.5 ± 1.3^a
F值	?	217.015	20.590	168.766
P值	?	< 0.001	< 0.001	< 0.001

组别	例数	NLRP3	Caspase-1
LPS组	6	2.04 ± 0.25	0.62 ± 0.07
LPS +抑制剂组	6	2.04 ± 0.27	0.51 ± 0.09^a
阴性对照组	6	1.49 ± 0.28^ab	0.50 ± 0.09^a
F值	?	8.723	3.905
P值	?	0.003	0.043

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