Echinoside Inhibits IL-1β-induced Chondrocytes Iron Death Through Nrf2/HO-1 Pathway

doi:10.19586/j.2095-2341.2025.0043

Current Biotechnology ›› 2025, Vol. 15 ›› Issue (4): 720-725.DOI: 10.19586/j.2095-2341.2025.0043

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Echinoside Inhibits IL-1β-induced Chondrocytes Iron Death Through Nrf2/HO-1 Pathway

Chuancai LIANG¹(), Bo QIU²()

^1.Department of Emergency，Wuhan University Renmin Hospital，Wuhan 430060，China
^2.Department of Orthopedics，Wuhan University Renmin Hospital，Wuhan 430060，China

Received:2025-03-27 Accepted:2025-05-07 Online:2025-07-25 Published:2025-09-08
Contact: Bo QIU

松果菊苷通过Nrf2/HO-1通路抑制IL-1β诱导的软骨细胞铁死亡

梁传财¹(), 邱波²()

^1.武汉大学人民医院急诊科，武汉 430060
^2.武汉大学人民医院关节外科，武汉 430060

通讯作者: 邱波
作者简介:梁传财 E-mail: 2020283020228@whu.edu.cn；
基金资助:
国家自然科学基金项目(81071494)

Abstract

Abstract:

To investigate the protective mechanism of echinacoside （ECH） in interleukin-1β （IL-1β）-induced chondrocyte iron death. Wistar mammary rat chondrocytes were extracted， and the experiment was divided into 5 groups： control group， IL-1β group， ECH+IL-1β group， ECH+IL-1β+Brusatol （Nrf2 inhibitor） group and ECH+IL-1β+ZnPP （HO-1 inhibitor） group. Quantitative PCR （qPCR） and Western Blot were used to detect mRNA and protein expression nuclear factor E2-related factor 2 （Nrf2）， heme oxygenase-1 （HO-1）， esteroyl coenzyme A synthase long-chain family member 4 （ACSL4） and glutathione peroxidase 4 （GPX4）， respectively. The results showed that the mRNA and protein expression of Nrf2， HO-1 and GPX4 were significantly reduced in the IL-1β group， and the mRNA and protein expression of ACSL4 were significantly enhanced compared with the control group. The mRNA and protein expression of cartilage Nrf2， HO-1 and GPX4 were significantly decreased and that of ACSL4 was significantly enhanced by the addition of the Nrf2 inhibitor Brusatol or the HO-1 inhibitor ZnPP in the ECH group. The results of Fe²⁺ Assay Kit showed that the chondrocyte Fe²⁺ content was significantly higher in the IL-1β group compared to the control group. The chondrocyte Fe²⁺ content was significantly lower in the ECH group （ECH+IL-1β） compared to the IL-1β group， while it was significantly higher in the ECH group after the addition of the Nrf2 inhibitor Brusatol or the HO-1 inhibitor ZnPP. The results suggest that echinoside can significantly improve IL-1β-induced chondrocyte iron death through Nrf2/HO-1 pathway.

Key words: osteoarthritis, iron death, echinoside, nuclear factor E2-related factor 2, heme oxygenase-1

摘要：

探讨了松果菊苷（echinacoside，ECH）对白细胞介素-1β（interleukin-1β， IL-1β）诱导的软骨细胞铁死亡的影响及其作用机制。提取Wistar大鼠软骨细胞，实验分为对照组、IL-1β组、ECH+IL-1β组、ECH+IL-1β+Brusatol(Nrf2抑制剂)组和ECH+IL-1β+ZnPP（HO-1抑制剂）5组。采用qPCR和Western Blot技术检测核因子E2相关因子2（nuclear factor-erythroid 2-related factor-2，Nrf2）、血红素氧合酶-1（heme oxygenase-1，HO-1）、酯酰辅酶A合成酶长链家族成员4（acyl-CoA synthetase long-chain familymember4，ACSL4）和谷胱甘肽过氧化酶4（glutathione peroxidase 4，GPX4）mRNA和蛋白表达量。结果显示，与对照组相比，IL-1β组中Nrf2、HO-1、GPX4的mRNA和蛋白表达量明显降低，ACSL4的mRNA和蛋白表达量明显升高。ECH组（ECH+IL-1β）中Nrf2、HO-1、GPX4的mRNA和蛋白表达量明显升高，ACSL4 的mRNA和蛋白表达量明显降低。在ECH组加入Nrf2抑制剂Brusatol或HO-1抑制剂ZnPP后，软骨细胞Nrf2、HO-1和GPX4的mRNA和蛋白表达量明显下降，ACSL4的mRNA和蛋白表达量明显升高。Fe²⁺检测试剂盒结果显示：与对照组相比，IL-1β组软骨细胞Fe²⁺含量明显升高；与IL-1β组相比，ECH组（ECH+IL-1β）软骨细胞Fe²⁺的含量明显降低，而在加入Nrf2抑制剂Brusatol或HO-1抑制剂ZnPP后软骨细胞Fe²⁺的含量明显升高。结果表明，松果菊苷通过Nrf2/HO-1通路可明显改善IL-1β诱导的软骨细胞铁死亡。

关键词: 骨关节炎, 铁死亡, 松果菊苷, 核因子E2相关因子2, 血红素氧合酶-1

CLC Number:

Q291

Chuancai LIANG, Bo QIU. Echinoside Inhibits IL-1β-induced Chondrocytes Iron Death Through Nrf2/HO-1 Pathway[J]. Current Biotechnology, 2025, 15(4): 720-725.

梁传财, 邱波. 松果菊苷通过Nrf2/HO-1通路抑制IL-1β诱导的软骨细胞铁死亡[J]. 生物技术进展, 2025, 15(4): 720-725.

Figures/Tables 5

References 24

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目的基因	引物序列（5′→3′）
GAPDH	F: 5′-ACAGCAACAGGGTGGTGGAC-3′
	R: 5′-TTTGAGGGTGCAGCGAACTT-3′
HO-1	F:5′-ACATCGACAGCCCCACCAAGTTCAA-3′
	R:5′-CTGACGAAGTGACGCCATCTGTGAG-3′
Nrf2	F:5′-TCCTCCTCGCTGGAAAAAGAA-3′
	R:5′-AATGTGCTGGCTGTGCTTTA-3′
GPX4	F:5′-ACCTGGACGCCAAAGTCCTA-3′
	R:5′-GTGACGATGCACACGAAACC-3′
ACSL4	F:5′-CAAGGGAGGCTTTAGGCGAC-3′
	R:5′-GCTCACTAAGGACGTCACCG-3′

目的基因	引物序列（5′→3′）
GAPDH	F: 5′-ACAGCAACAGGGTGGTGGAC-3′
	R: 5′-TTTGAGGGTGCAGCGAACTT-3′
HO-1	F:5′-ACATCGACAGCCCCACCAAGTTCAA-3′
	R:5′-CTGACGAAGTGACGCCATCTGTGAG-3′
Nrf2	F:5′-TCCTCCTCGCTGGAAAAAGAA-3′
	R:5′-AATGTGCTGGCTGTGCTTTA-3′
GPX4	F:5′-ACCTGGACGCCAAAGTCCTA-3′
	R:5′-GTGACGATGCACACGAAACC-3′
ACSL4	F:5′-CAAGGGAGGCTTTAGGCGAC-3′
	R:5′-GCTCACTAAGGACGTCACCG-3′

组别	Nrf2	HO-1	GPX4	ACSL4
对照组	0.847±0.094^*	0.861±0.109^*	0.836±0.101^*	0.587±0.071^*
IL-1β组	0.381±0.049	0.458±0.027	0.402±0.047	1.373±0.068
ECH+IL-1β组	0.612±0.034^*	0.657±0.026^*	0.632±0.092^*	0.581±0.076^*
ECH+IL-1β+Brusatol组	0.349±0.055^#	0.367±0.066^#	0.415±0.071^#	0.911±0.036^#
ECH+IL-1β+ZnPP组	0.363±0.015^#	0.381±0.014^#	0.323±0.014^#	0.935±0.095^#
F值	45	140	25	59
P值	<0.05	<0.05	<0.05	<0.05

组别	Nrf2	HO-1	GPX4	ACSL4
对照组	0.847±0.094^*	0.861±0.109^*	0.836±0.101^*	0.587±0.071^*
IL-1β组	0.381±0.049	0.458±0.027	0.402±0.047	1.373±0.068
ECH+IL-1β组	0.612±0.034^*	0.657±0.026^*	0.632±0.092^*	0.581±0.076^*
ECH+IL-1β+Brusatol组	0.349±0.055^#	0.367±0.066^#	0.415±0.071^#	0.911±0.036^#
ECH+IL-1β+ZnPP组	0.363±0.015^#	0.381±0.014^#	0.323±0.014^#	0.935±0.095^#
F值	45	140	25	59
P值	<0.05	<0.05	<0.05	<0.05

组别	Nrf2	HO-1	GPX4	ACSL4
对照组	1.153±0.107^*	1.014±0.141^*	1.019±0.041^*	1.162±0.151^*
IL-1β组	0.443±0.070	0.414±0.017	0.436±0.026	3.236±0.252
ECH+IL-1β组	0.906±0.032^*	0.793±0.051^*	0.782±0.094^*	1.64±0.086^*
ECH+IL-1β+Brusatol组	0.656±0.075^#	0.618±0.053^#	0.611±0.012^#	2.024±0.246^#
ECH+IL-1β+ZnPP组	0.613±0.021^#	0.585±0.018^#	0.619±0.031^#	2.564±0.116^#
F值	48	77	51	91
P值	<0.05	<0.05	<0.05	<0.05

Echinoside Inhibits IL-1β-induced Chondrocytes Iron Death Through Nrf2/HO-1 Pathway

松果菊苷通过Nrf2/HO-1通路抑制IL-1β诱导的软骨细胞铁死亡

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组别	Fe²⁺含量/(μmol·mL^-1)	细胞增殖率/%
对照组	110.1±7.1^*	81.3±5.3^*
IL-1β组	424.3±30.4	53.4±7.1
ECH+IL-1β组	266.0±12.3^*	75.9±4.7^*
ECH+IL-1β+Brusatol组	371.2±25.1^#	60.2±3.2^#
ECH+IL-1β+ZnPP组	362.5±21.7^#	57.5±6.2^#
F值	103	68
P值	<0.05	<0.05

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