Naa20 alleviated myocardial injury in sepsis by inhibiting AIM2 expression

doi:10.11958/20230338

Abstract

Abstract:

Objective To explore the regulatory effect and mechanism of Naa20 on lipopolysaccharide (LPS) induced myocardial injury in sepsis. Methods Twelve male C57BL/6J mice were divided into the control group and the myocardial injury model group (n=6 in each group). Mice of the myocardial injury model group were treated with a single intravenous injection of LPS, while mice of the control group were injected with the same dose of PBS. Cardiac function of mice in each group was evaluated by cardiac Doppler ultrasonography 12 h after LPS injection. The serum content of CK-MB was detected by enzyme-linked immunosorbent assay (ELISA). Hematoxylin-eosin staining (HE) was used to evaluate myocardial injury in mice. Cardiomyocytes from SD rats were isolated and cultured in vitro and divided into the control group and the LPS group. Mitochondrial membrane potential and reactive oxygen species (ROS) release in cardiomyocytes were detected by fluorescent probe. The protein expression levels of AIM2, GSDMD, activated GSDMD (GSDMD-N), IL-1β, activated IL-1β (p17) and Naa20 were detected by Western blot assay. The content of IL-1β in supernatant was determined by ELISA. The expression level of Naa20 in cardiomyocytes was detected by quantitative real-time PCR (qPCR). H9c2 cells were randomly divided into the control group, the Naa20 group, the LPS group and the Naa20+LPS group. Methyl thiazolyl tetrazolium (MTT) assay and lactate dehydrogenase (LDH) assay were used to detect the viability of H9c2 cells. The interaction between Naa20 and AIM2 was detected by immunoprecipitation. Results Compared with the control group, the left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (FS) were decreased in the myocardial injury model group (P<0.01), the serum content of CK-MB was increased (P<0.01), and HE staining showed pathological changes in heart of mice. In vitro, compared with the control group, mitochondrial membrane potential was decreased, while ROS production, expressions of AIM2, GSDMD, GSDMD-N, IL-1β, p17 and Naa20 and the content of IL-1β in supernatant were significantly increased in the LPS group (P<0.05). Results of immunocoprecipitation confirmed that Naa20 interacted with AIM2. Compared with the LPS group, the viability of H9c2 cells and the acetylation of AIM2 were increased, while the protein expression levels of AIM2, GSDMD, GSDMD-N and the content of IL-1β in supernatant were decreased in the Naa20+LPS group (P<0.05). Conclusion Naa20 can regulate AIM2-mediacted pyroptosis by inhibiting AIM2 expression, thereby alleviating sepsis-induced myocardial injury.

Key words: pyroptosis, Naa20, AIM2, myocardial injury

CLC Number:

R631.3

Figures/Tables 14

References 22

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基因	引物序列（5′→3′）	产物大小/bp
Naa20	上游：GACGACCTGTTCCGCTTCAA 下游：TCTGCTTTGCCCATAACCAA	148
GAPDH	上游：CATGAGAAGTATGACAACAGCT 下游：AGTCCTTCCACGATACCAAAGT	197

基因	引物序列（5′→3′）	产物大小/bp
Naa20	上游：GACGACCTGTTCCGCTTCAA 下游：TCTGCTTTGCCCATAACCAA	148
GAPDH	上游：CATGAGAAGTATGACAACAGCT 下游：AGTCCTTCCACGATACCAAAGT	197

组别	LVEF	FS/%	CK-MB/（ng/L）
对照组	0.70±0.04	37.93±3.48	345.84±250.09
模型组	0.40±0.05	18.88±3.05	7 573.23±1 194.54
t	10.765^＊＊	10.084^＊＊	14.506^＊＊

组别	LVEF	FS/%	CK-MB/（ng/L）
对照组	0.70±0.04	37.93±3.48	345.84±250.09
模型组	0.40±0.05	18.88±3.05	7 573.23±1 194.54
t	10.765^＊＊	10.084^＊＊	14.506^＊＊

组别		AIM2		GSDMD		GSDMD-N
对照组		1.00±0.00		1.00±0.00		1.00±0.00
LPS处理组		3.65±1.49		2.47±0.16		1.57±0.21
t		3.103^＊		15.777^＊＊		4.637^＊
组别	IL-1β		p17		IL-1β/（ng/L）
对照组	1.00±0.00		1.00±0.00		1.07±0.35
LPS处理组	2.76±0.38		2.52±0.27		30.24±2.53
t	7.958^＊＊		9.782^＊＊		19.759^＊＊