The effect of esculin on hypoxia/reoxygenation induced myocardial cell injury by regulating HMGB1/RAGE signaling pathway

doi:10.11958/20251272

Abstract

Abstract:

Objective To explore the effect of esculin (ESCL) on hypoxia/reoxygenation (H/R)-induced myocardial cell injury by regulating high mobility group B1 (HMGB1)/receptor of advanced glycation endproducts (RAGE) signaling pathway. Methods H9c2 cells were divided into the control group, the H/R group, the ESCL-low group, the ESCL-medium group and the ESCL-high group (ESCL-L, M, H, 0.4, 0.8, 1.6 mmol/L), the ESCL-H+pcDNA-NC group and the ESCL-H+pcDNA-HMGB1 group. The AnnexinV FITC staining method was used to detect the apoptosis rate of H9c2 cells. Enzyme linked immunosorbent assay was used to measure interleukin (IL)-18, IL-1β, tumor necrosis factor (TNF)-α, the malondialdehyde (MDA), lactate dehydrogenase (LDH) contents and the superoxide dismutase (SOD) activity. The DCHF-DA fluorescent probe was performed to detect active oxygen (ROS) fluorescence intensity. Immunoblotting was performed to detect B-cell lymphoma-2 (Bcl-2), Bcl-2 associated X protein (Bax), HMGB1 and RAGE proteins. Results Compared with the control group，the H/R group showed conspicuously higher apoptosis rate, IL-18, IL-1β, TNF-α, MDA, LDH, ROS fluorescence intensity, Bax, HMGB1 and RAGE proteins in H9c2 cells, and conspicuously lower Bcl-2 protein and SOD activity (P<0.05). Compared with the H/R group, the ESCL-L, ESCL-M and ESCL-H groups showed a gradual decrease in cell apoptosis rate, IL-18, IL-1β, TNF-α, MDA, LDH and ROS fluorescence intensity, Bax, HMGB1 and RAGE proteins with increasing ESCL concentration, and a gradual increase in Bcl-2 protein and SOD activity (P<0.05). Overexpression of HMGB1 reversed the protective effect of ESCL on H/R induced cardiomyocyte injury. Conclusion ESCL may reduce H/R induced cardiomyocyte injury by inhibiting HMGB1/RAGE pathway.

Key words: myocytes, cardiac, aesculi, HMGB1 protein, receptor for advanced glycation end products, apoptosis, hypoxia, reoxygenation

CLC Number:

R542.2

Figures/Tables 8

References 23

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组别	细胞凋亡率/%	Bax/GAPDH	Bcl-2/GAPDH
Control组	4.78±0.56	0.12±0.03	1.13±0.10
H/R组	45.52±4.73^a	1.17±0.12^a	0.16±0.02^a
ESCL-L组	39.48±3.21^b	0.85±0.16^b	0.28±0.04^b
ESCL-M组	27.64±2.65^bc	0.55±0.15^bc	0.42±0.06^bc
ESCL-H组	12.26±1.37^bcd	0.21±0.13^bcd	0.63±0.07^bcd
ESCL-H+pcDNA-NC组	12.27±1.36	0.22±0.12	0.62±0.06
ESCL-H+pcDNA-HMGB1组	30.01±3.64^e	0.70±0.18^e	0.33±0.04^e
F	171.311^＊	49.907^＊	170.724^＊

组别	细胞凋亡率/%	Bax/GAPDH	Bcl-2/GAPDH
Control组	4.78±0.56	0.12±0.03	1.13±0.10
H/R组	45.52±4.73^a	1.17±0.12^a	0.16±0.02^a
ESCL-L组	39.48±3.21^b	0.85±0.16^b	0.28±0.04^b
ESCL-M组	27.64±2.65^bc	0.55±0.15^bc	0.42±0.06^bc
ESCL-H组	12.26±1.37^bcd	0.21±0.13^bcd	0.63±0.07^bcd
ESCL-H+pcDNA-NC组	12.27±1.36	0.22±0.12	0.62±0.06
ESCL-H+pcDNA-HMGB1组	30.01±3.64^e	0.70±0.18^e	0.33±0.04^e
F	171.311^＊	49.907^＊	170.724^＊

组别	IL-18	IL-1β	TNF-α
Control组	73.51±7.36	83.14±6.38	105.64±10.53
H/R组	259.57±29.41^a	266.54±26.95^a	330.85±30.23^a
ESCL-L组	225.69±21.39^b	230.02±21.54^b	285.12±29.56^b
ESCL-M组	176.46±17.96^bc	186.77±18.95^bc	230.04±26.21^bc
ESCL-H组	123.31±13.52^bcd	125.83±12.48^bcd	156.28±16.43^bcd
ESCL-H+pcDNA-NC组	124.02±13.05	126.46±12.76	157.30±15.92
ESCL-H+pcDNA-HMGB1组	195.97±18.85^e	215.97±20.95^e	259.42±27.45^e
F	73.924^＊	78.787^＊	71.079^＊

组别	IL-18	IL-1β	TNF-α
Control组	73.51±7.36	83.14±6.38	105.64±10.53
H/R组	259.57±29.41^a	266.54±26.95^a	330.85±30.23^a
ESCL-L组	225.69±21.39^b	230.02±21.54^b	285.12±29.56^b
ESCL-M组	176.46±17.96^bc	186.77±18.95^bc	230.04±26.21^bc
ESCL-H组	123.31±13.52^bcd	125.83±12.48^bcd	156.28±16.43^bcd
ESCL-H+pcDNA-NC组	124.02±13.05	126.46±12.76	157.30±15.92
ESCL-H+pcDNA-HMGB1组	195.97±18.85^e	215.97±20.95^e	259.42±27.45^e
F	73.924^＊	78.787^＊	71.079^＊

组别	MDA/（μmol/L）	LDH/（U/L）	SOD/（U/L）	ROS荧光强度
Control组	1.63±0.53	65.26±5.72	37.12±5.43	102.53±5.35
H/R组	15.37±1.45^a	130.31±12.05^a	12.06±3.12^a	345.06±23.41^a
ESCL-L组	11.68±1.06^b	110.05±13.24^b	19.64±1.91^b	297.75±28.54^b
ESCL-M组	8.25±0.78^bc	90.72±9.12^bc	26.37±2.34^bc	250.72±25.43^bc
ESCL-H组	5.09±0.61^bcd	70.53±7.78^bcd	33.26±2.69^bcd	206.64±23.10^bcd
ESCL-H+pcDNA-NC组	5.10±0.53	71.37±8.01	34.14±3.08	205.79±23.04
ESCL-H+pcDNA-HMGB1组	13.93±3.67^e	110.51±12.16^e	22.92±2.43^e	271.09±21.01^e
F	60.222^＊	36.889^＊	47.862^＊	72.366^＊