Pinocembrin alleviates BEAS-2B cell damage induced by chronic intermittent hypoxia through inhibiting autophagy

doi:10.16138/j.1673-6087.2024.02.05

Abstract

Abstract:

Objective To investigate the effect of pinocembrin (PIN) on BEAS-2B human bronchial epithelial cell injury induced by chronic intermittent hypoxia (CIH) and its possible regulatory mechanism. Methods The optimal CIH intervention duration and PIN treatment concentration were determined by cell counting kit 8 (CCK8) in BEAS-2B cells. BEAS-2B cells were divided into control group (CON group), CIH group and CIH+PIN group. The proliferation activity of BEAS-2B cells in different treatment groups was detected by CCK8. Western blotting (WB) was used to detect the protein expression levels of hypoxia-inducible factor 1α (HIF-1α), apoptosis related gene [cleaved caspase 3, B cell lymphoma 2 (Bcl-2), Bcl-2 associated X protein (Bax)] and autophagy related genes [light chain 3(LC3)-Ⅱ, beclin-1, P62]. After adding the autophagy inhibitor 3-methyladenine (3-MA), the cells were divided into five groups: CON group, CIH group, CIH+3-MA group, CIH+PIN group, and CIH+3-MA+PIN group, and the expression levels of HIF-1α, apoptosis and autophagy related genes in each group were detected. Results CCK8 showed that the proliferation activity of BEAS-2B cells significantly decreased between 24-48 h with the prolongation of hypoxia time. PIN could up-regulate the proliferative activity of BEAS-2B cells under CIH. The results of WB showed that compared with CON group, the protein expressions of HIF-1α, Bax, LC3-Ⅱ and beclin-1 were increased in CIH group (P<0.05). The expression of P62 and anti-apoptosis gene Bcl-2 protein was significantly down-regulated (P<0.05). After adding 3-MA, the protein expression level of HIF-1α was reduced, autophagy and apoptosis were inhibited. The same trend was observed after adding PIN. Conclusions PIN can reduce CIH-induced BEAS-2B cell damage and exert a protective effect. The mechanism may be related to down-regulating autophagy level and inhibiting cell apoptosis.

Key words: Pinocembrin, Chronic intermittent hypoxia, Autophagy, Apoptosis

CLC Number:

R563.5

HE Yanjie, HE Meijuan, WANG Yun, ZHU Chunxue, HUANG Hanpeng. Pinocembrin alleviates BEAS-2B cell damage induced by chronic intermittent hypoxia through inhibiting autophagy[J]. Journal of Internal Medicine Concepts & Practice, 2024, 19(02): 115-120.

Figures/Tables 7

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项目	CON组	CIH组	CIH+PIN组	F	P
HIF-1α	1	1.236±0.019¹⁾	0.959±0.022¹⁾²⁾	97.59	<0.000 1
裂解的胱天蛋白酶3	1	1.278±0.025¹⁾	0.906±0.049¹⁾²⁾	31.76	<0.000 6
Bax	1	1.371±0.029¹⁾	1.024±0.031¹⁾²⁾	138.60	<0.000 1
Bcl-2	1	0.875±0.024¹⁾	1.133±0.047¹⁾²⁾	9.20	<0.05
P62	1	0.807±0.016¹⁾	1.163±0.039¹⁾²⁾	20.88	<0.002
beclin-1	1	1.340±0.039¹⁾	0.874±0.062¹⁾²⁾	44.61	<0.000 3
LC3-Ⅱ	1	1.291±0.027¹⁾	0.912±0.086¹⁾²⁾	8.137	<0.05

项目	CON组	CIH组	CIH+3-MA组	CIH+PIN组	CIH+3-MA+PIN组	F	P
HIF-1α	1	1.432±0.039¹⁾	1.221±0.039¹⁾²⁾	1.267±0.040¹⁾²⁾	1.200±0.047¹⁾²⁾	49.96	<0.000 1
裂解的胱天蛋白酶3	1	1.756±0.069¹⁾	1.427±0.087¹⁾²⁾	1.507±0.105¹⁾	1.439±0.093¹⁾²⁾	20.92	<0.000 1
Bcl-2	1	0.582±0.030¹⁾	0.755±0.035¹⁾²⁾	0.890±0.033¹⁾²⁾	0.973±0.054¹⁾²⁾	44.73	<0.000 1
LC3-Ⅱ	1	1.445±0.020¹⁾	1.038±0.126¹⁾²⁾	1.063±0.137¹⁾²⁾	0.775±0.134¹⁾²⁾	13.68	<0.000 5
beclin-1	1	1.419±0.020¹⁾	1.129±0.058¹⁾²⁾	1.158±0.028¹⁾²⁾	1.080±0.040¹⁾²⁾	26.64	<0.000 1
P62	1	0.605±0.015¹⁾	0.879±0.023¹⁾²⁾	0.976±0.019¹⁾²⁾	1.074±0.030¹⁾²⁾	100.40	<0.000 1