Effects of matrine on inflammation, oxidative stress and wound healing in atopic dermatitis

doi:10.11958/20231689

Abstract

Abstract:

Objective To evaluate the effects of matrine on the inflammatory, oxidative and migratory responses of HaCaT keratinocytes in an in vitro atopic dermatitis (AD) model. Methods HaCaT cells were divided into the control group (no treatment) and the model group (0.05, 0.1 and 0.2 mmol/L). HaCaT cells in the model group were stimulated with 10 μg/L tumor necrosis factor-α (TNF-α)/interferon-γ (IFN-γ) for 24 h. Cells in the model group were pretreated with 0.05, 0.1 and 0.2 mmol/L matrine for 1 h and then stimulated with 10 μg/L TNF-α/IFN-γ for 24 h. MTT assay was used to assess the cytotoxicity of matrine. Enzyme-linked immunosorbent (ELISA) assay was performed to determine levels of cytokines (IL-6, IL-8 and IL-1β) and chemokines (TARC, MDC and RANTES). mRNA expression levels of antioxidant genes were detected by real-time quantitative PCR (RT-qPCR). Cell migration was analyzed by scratch closure assay. Immunohistochemistry was commended to analyze the nuclear translocation of NF-κB p65. Western blot assay was used to detect the phosphorylation levels of p38, ERK and p65 proteins. Results Compared with the model group, matrine 0.1 and 0.2 mmol/L significantly decreased the expression levels of IL-6, IL-1β, IL-8 and chemokines, and matrine significantly enhanced the expression levels of superoxide dismutase-1 (SOD1), SOD2, catalase (CAT) and glutathione peroxidase (GPx), and promoted scratch wound healing (P<0.05). In addition, matrine 0.1 and 0.2 mmol/L inhibited the phosphorylation levels of p38, ERK and p65 proteins and the nuclear translocation of p65. Conclusion Matrine possesses anti-inflammatory and antioxidant properties, and stimulates wound closure in eratinocyte AD model, which may be related to the inhibition of the activation of MAPK and NF-κB pathways.

Key words: matrine, dermatitis, atopic, inflammation, NF-kappa B, oxidative stress

CLC Number:

R758.2

Figures/Tables 8

References 19

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基因名称	引物序列（5′→3′）	产物大小/bp
SOD1	上游：ACGGTGGGCCAAAGGATGAA	151
	下游：TCATGGACCACCAGTGTGCG	151
SOD2	上游：AGAAGCACAGCCTCCCCGAC	152
	下游：GGCCAACGCCTCCTGGTACT	152
CAT	上游：CCAACAGCTTTGGTGCTCCG	180
	下游：GGCCGGCAATGTTCTCACAC	180
GPx	上游：TCGGTGTATGCCTTCTCGGC	150
	下游：CCGCTGCAGCTCGTTCATCT	150
GAPDH	上游：TTGGTATCGTGGAAGGACTCA	270
	下游：TGTCATCATATTTGGCAGGTTT	270

基因名称	引物序列（5′→3′）	产物大小/bp
SOD1	上游：ACGGTGGGCCAAAGGATGAA	151
	下游：TCATGGACCACCAGTGTGCG	151
SOD2	上游：AGAAGCACAGCCTCCCCGAC	152
	下游：GGCCAACGCCTCCTGGTACT	152
CAT	上游：CCAACAGCTTTGGTGCTCCG	180
	下游：GGCCGGCAATGTTCTCACAC	180
GPx	上游：TCGGTGTATGCCTTCTCGGC	150
	下游：CCGCTGCAGCTCGTTCATCT	150
GAPDH	上游：TTGGTATCGTGGAAGGACTCA	270
	下游：TGTCATCATATTTGGCAGGTTT	270

组别	细胞因子
组别	IL-6		IL-1β		IL-8
对照组	0.82±0.14		31.5±3.3		9.7±1.8
模型组	6.73±0.57^a		115.2±6.5^a		78.8±6.1^a
0.05 mmol/L组	6.48±0.55		107.8±6.7		76.4±5.7
0.1 mmol/L组	5.51±0.42^b		72.3±4.8^b		49.3±3.8^b
0.2 mmol/L组	4.46±0.35^b		36.6±3.6^b		17.6±2.2^b
F	91.210^＊＊		169.000^＊＊		168.100^＊＊
组别		趋化因子
组别		MDC		TARC		RANTES
对照组		65±7		19±3		11±2
模型组		341±15^a		443±20^a		151±17^a
0.05 mmol/L组		337±18		431±24		147±19
0.1 mmol/L组		208±11^b		346±18^b		79±13^b
0.2 mmol/L组		110±13^b		277±15^b		38±7^b
F		271.500^＊＊		291.300^＊＊		68.470^＊＊

组别	细胞因子
组别	IL-6		IL-1β		IL-8
对照组	0.82±0.14		31.5±3.3		9.7±1.8
模型组	6.73±0.57^a		115.2±6.5^a		78.8±6.1^a
0.05 mmol/L组	6.48±0.55		107.8±6.7		76.4±5.7
0.1 mmol/L组	5.51±0.42^b		72.3±4.8^b		49.3±3.8^b
0.2 mmol/L组	4.46±0.35^b		36.6±3.6^b		17.6±2.2^b
F	91.210^＊＊		169.000^＊＊		168.100^＊＊
组别		趋化因子
组别		MDC		TARC		RANTES
对照组		65±7		19±3		11±2
模型组		341±15^a		443±20^a		151±17^a
0.05 mmol/L组		337±18		431±24		147±19
0.1 mmol/L组		208±11^b		346±18^b		79±13^b
0.2 mmol/L组		110±13^b		277±15^b		38±7^b
F		271.500^＊＊		291.300^＊＊		68.470^＊＊

组别	SOD1	SOD2	CAT	GPx
对照组	1.00±0.04	1.00±0.05	1.00±0.04	1.00±0.03
模型组	0.51±0.02^a	0.42±0.07^a	0.27±0.03^a	0.30±0.04^a
0.05 mmol/L组	0.53±0.03	0.44±0.09	0.30±0.05	0.34±0.02
0.1 mmol/L组	0.88±0.06^b	5.85±0.96^b	0.52±0.06^b	0.56±0.06^b
0.2 mmol/L组	1.25±0.09^b	12.34±1.28^b	0.84±0.08^b	1.07±0.09^b
F	102.800^＊＊	156.500^＊＊	105.500^＊＊	135.000^＊＊