基于STING/ACSL4通路探讨复元活血汤抑制膝骨关节炎大鼠软骨细胞铁死亡的作用机制

doi:10.11958/20261146

天津医药 ›› 2026, Vol. 54 ›› Issue (10): 1009-1017.doi: 10.11958/20261146

• 实验研究 • 下一篇

基于STING/ACSL4通路探讨复元活血汤抑制膝骨关节炎大鼠软骨细胞铁死亡的作用机制

冯丽娜¹(), 阎亮²^,^△(), 薛水娜³, 王上增³

¹ 河南省中医院（河南中医药大学第二附属医院）关节病一区（邮编 450000）
² 河南省中医院（河南中医药大学第二附属医院）关节病二区（邮编 450000）
³ 河南省中医院（河南中医药大学第二附属医院）关节科（邮编 450000）

收稿日期:2026-04-30 修回日期:2026-05-29 出版日期:2026-10-15 发布日期:2026-10-10
通讯作者: △E-mail：33946314@qq.com
作者简介:冯丽娜（1984），女，主治医师，主要从事关节运动损伤方面研究。E-mail：flnlina@126.com
基金资助:
河南省中青年卫生健康科技创新人才培养项目(LJRC2024020);河南省中医药科学研究专项课题(2023ZYZD06)

Mechanism of Fuyuan Huoxue decoction in inhibiting chondrocyte ferroptosis of rats with knee osteoarthritis via the STING/ACSL4 pathway

FENG Lina¹(), YAN Liang²^,^△(), XUE Shuina³, WANG Shangzeng³

¹ Department Ⅰ of Joint Diseases, Henan Province Hospital of Traditional Chinese Medicine, Zhengzhou 450000, China
² Department II of Joint Diseases, Henan Province Hospital of Traditional Chinese Medicine, Zhengzhou 450000, China
³ Department of Joints, Henan Province Hospital of Traditional Chinese Medicine, Zhengzhou 450000, China

Received:2026-04-30 Revised:2026-05-29 Published:2026-10-15 Online:2026-10-10
Contact: △E-mail：33946314@qq.com

冯丽娜, 阎亮, 薛水娜, 王上增. 基于STING/ACSL4通路探讨复元活血汤抑制膝骨关节炎大鼠软骨细胞铁死亡的作用机制[J]. 天津医药, 2026, 54(10): 1009-1017.

FENG Lina, YAN Liang, XUE Shuina, WANG Shangzeng. Mechanism of Fuyuan Huoxue decoction in inhibiting chondrocyte ferroptosis of rats with knee osteoarthritis via the STING/ACSL4 pathway[J]. Tianjin Medical Journal, 2026, 54(10): 1009-1017.

摘要/Abstract

摘要：

目的基于干扰素基因刺激因子（STING）/长链酰基辅酶A合成酶家族成员4（ACSL4）通路探讨复元活血汤（FHD）抑制膝骨关节炎（KOA）大鼠软骨细胞铁死亡的作用机制。方法将SD大鼠随机分为假手术（Sham）组、模型（KOA）组、FHD低剂量（FHD-L）组、FHD高剂量（FHD-H）组、塞来昔布阳性对照（PC）组及FHD-H+STING激活剂（FHD-H+DMXAA）组，每组12只。采用改良Hulth法建立KOA模型。造模成功后各组分别给予相应药物灌胃或尾静脉注射干预，连续8周。采用Lequesne MG评分评估膝关节功能；HE染色观察软骨组织形态；番红O-固绿染色结合OARSI评分评估软骨退变程度；酶联免疫吸附试验检测软骨组织中白细胞介素（IL）-1β、肿瘤坏死因子（TNF）-α、基质金属蛋白酶（MMP）-13及血小板反应蛋白解整合素金属肽酶-5（ADAMTS-5）水平；生化试剂盒测定软骨组织Fe²⁺、丙二醛（MDA）、谷胱甘肽（GSH）及超氧化物歧化酶（SOD）水平；DCFH-DA探针法检测软骨组织活性氧（ROS）水平；普鲁士蓝染色观察铁沉积；透射电镜观察软骨细胞超微结构；免疫组化法检测软骨组织谷胱甘肽过氧化物酶4（GPX4）、溶质载体家族7成员11（SLC7A11）、环氧化酶-2（COX-2）蛋白表达；Western blot法检测软骨组织STING及ACSL4蛋白表达。结果与Sham组相比，KOA组Lequesne MG评分，OARSI评分，软骨组织炎症及基质降解因子（IL-1β、TNF-α、MMP-13、ADAMTS-5），铁死亡相关指标（Fe²⁺、MDA、ROS），普鲁士蓝染色阳性面积比，COX-2阳性表达面积及STING、ACSL4蛋白表达均显著升高（P<0.05），而GSH、SOD水平及GPX4、SLC7A11阳性表达面积均显著降低（P<0.05）；HE染色、番红O-固绿染色及透射电镜显示KOA组软骨结构严重破坏，软骨基质丢失，线粒体萎缩、嵴断裂。与KOA组相比，FHD-L组、FHD-H组及PC组上述指标均有不同程度改善（P<0.05），其中FHD-H组效果优于FHD-L组（P<0.05）。与FHD-H组相比，FHD-H+DMXAA组上述指标的改善效应被显著逆转（P<0.05）。结论 FHD可能通过抑制STING/ACSL4通路减轻软骨细胞铁死亡，从而改善KOA大鼠的关节功能与软骨损伤。

关键词: 复元活血汤, 骨关节炎, 膝, 软骨细胞, 铁死亡, STING/ACSL4通路

Abstract:

Objective To investigate the mechanism by which Fuyuan Huoxue decoction (FHD) inhibiting chondrocyte ferroptosis in a rat model of knee osteoarthritis (KOA) via the stimulator of interferon genes (STING)/acyl-CoA synthetase long-chain family member 4 (ACSL4) pathway. Methods SD rats were randomly divided into six groups: the sham operation (Sham) group, the model (KOA) group, the low-dose FHD (FHD-L) group, the high-dose FHD (FHD-H) group, the positive control (PC, celecoxib) group and the FHD-H+STING activator (FHD-H+DMXAA) group, with 12 rats in each group. The KOA model was established using the modified Hulth method. After successful modeling, each group received corresponding interventions by intragastric administration or tail vein injection for 8 consecutive weeks. Knee joint function was assessed using the Lequesne MG score method. Cartilage morphology was observed by HE staining. Cartilage degeneration was evaluated by Safranin O-fast green staining combined with the OARSI score. Levels of interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), matrix metalloproteinase-13 (MMP-13) and the disintegrin and metalloproteinase with thrombospondin motifs-5 (ADAMTS-5) in cartilage tissue were measured by ELISA. Levels of Fe²⁺, malondialdehyde (MDA), glutathione (GSH) and superoxide dismutase (SOD) in cartilage tissue were determined using biochemical kits. The level of reactive oxygen species (ROS) in cartilage tissue was detected by DCFH-DA probe method. Iron deposition was observed by Prussian blue staining. Mitochondrial ultrastructure of chondrocytes was observed by transmission electron microscopy. The protein expressions of glutathione peroxidase 4 (GPX4), solute carrier family 7 member 11 (SLC7A11) and cyclooxygenase-2 (COX-2) in cartilage tissue were detected by immunohistochemistry. The protein expressions of STING and ACSL4 were detected by Western blot assay. Results Compared with the Sham group, Lequesne MG score, OARSI score, cartilage inflammatory and matrix degradation factors (IL-1β, TNF-α, MMP-13 and ADAMTS-5), ferroptosis-related indicators (Fe²⁺, MDA, ROS), Prussian blue staining positive area ratio, COX-2 positive expression area and STING and ACSL4 protein expression were significantly increased in the KOA group (P<0.05), while the levels of GSH and SOD, and the positive expression areas of GPX4 and SLC7A11 were significantly decreased (P<0.05). HE staining, Safranin O-fast green staining and transmission electron microscopy revealed severe cartilage structural damage, matrix loss, mitochondrial atrophy and cristae rupture in the KOA group. Compared with the KOA group, all the above indicators were improved to varying degrees in the FHD-L group, the FHD-H group and the PC group (P<0.05), with the FHD-H group showing better efficacy than the FHD-L group (P<0.05). Among them, the protective effects on the above indicators of the FHD-H group was better than that of the FHD-L group (P<0.05). Compared with the FHD-H group, the improvement effects of the above indicators in the FHD-H +DMXAA group were significantly reversed (P<0.05). Conclusion FHD may improve joint function and cartilage damage in KOA rats by inhibiting chondrocyte ferroptosis via suppressing the STING/ACSL4 pathway.

Key words: Fu Yuan Huo Xue Tang, osteoarthritis, knee, chondrocytes, ferroptosis, STING/ACSL4 pathway

中图分类号:

R684.3

图/表 12

参考文献 32

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组别	Lequesne MG评分（n=12）		OARSI评分（n=6）
组别	给药前	给药后	OARSI评分（n=6）
Sham组	0.08±0.02	0.22±0.08	1.03±0.52
KOA组	8.81±1.34^a	8.67±1.23^a	14.67±2.16^a
FHD-L组	8.69±1.22^a	5.92±0.71^b	10.83±1.94^b
FHD-H组	8.75±1.18^a	2.67±0.59^bc	5.17±1.47^bc
PC组	8.78±1.25^a	2.83±0.44^bc	5.83±1.60^bc
FHD-H+DMXAA组	8.72±1.16^a	7.08±0.86^d	12.50±2.07^d
F	118.941^＊＊	220.018^＊＊	54.232^＊＊

组别	Lequesne MG评分（n=12）		OARSI评分（n=6）
组别	给药前	给药后	OARSI评分（n=6）
Sham组	0.08±0.02	0.22±0.08	1.03±0.52
KOA组	8.81±1.34^a	8.67±1.23^a	14.67±2.16^a
FHD-L组	8.69±1.22^a	5.92±0.71^b	10.83±1.94^b
FHD-H组	8.75±1.18^a	2.67±0.59^bc	5.17±1.47^bc
PC组	8.78±1.25^a	2.83±0.44^bc	5.83±1.60^bc
FHD-H+DMXAA组	8.72±1.16^a	7.08±0.86^d	12.50±2.07^d
F	118.941^＊＊	220.018^＊＊	54.232^＊＊

组别	IL-1β/ （pg/mg）	TNF-α/ （pg/mg）	MMP-13/ （ng/mg）	ADAMTS-5/（ng/mg）
Sham组	12.47±2.15	8.23±1.56	3.42±0.58	2.15±0.42
KOA组	86.34±7.82^a	72.56±6.93^a	28.67±3.14^a	18.93±2.08^a
FHD-L组	58.26±6.14^b	48.37±5.25^b	18.42±2.56^b	12.67±1.85^b
FHD-H组	28.53±3.67^bc	22.18±3.42^bc	8.35±1.27^bc	5.28±0.96^bc
PC组	31.42±4.05^bc	24.56±3.78^bc	9.64±1.48^bc	6.12±1.03^bc
FHD-H+DMXAA组	67.38±6.52^d	56.29±5.84^d	22.53±2.87^d	15.47±1.92^d
F	158.243^＊＊	153.825^＊＊	115.575^＊＊	114.235^＊＊

组别	IL-1β/ （pg/mg）	TNF-α/ （pg/mg）	MMP-13/ （ng/mg）	ADAMTS-5/（ng/mg）
Sham组	12.47±2.15	8.23±1.56	3.42±0.58	2.15±0.42
KOA组	86.34±7.82^a	72.56±6.93^a	28.67±3.14^a	18.93±2.08^a
FHD-L组	58.26±6.14^b	48.37±5.25^b	18.42±2.56^b	12.67±1.85^b
FHD-H组	28.53±3.67^bc	22.18±3.42^bc	8.35±1.27^bc	5.28±0.96^bc
PC组	31.42±4.05^bc	24.56±3.78^bc	9.64±1.48^bc	6.12±1.03^bc
FHD-H+DMXAA组	67.38±6.52^d	56.29±5.84^d	22.53±2.87^d	15.47±1.92^d
F	158.243^＊＊	153.825^＊＊	115.575^＊＊	114.235^＊＊

组别		Fe²⁺/（μmol/g）	MDA/（nmol/g）		ROS（荧光强度）
Sham组		0.28±0.05	1.32±0.21		100.00±8.56
KOA组		1.86±0.17^a	5.78±0.52^a		286.54±18.63^a
FHD-L组		1.28±0.14^b	4.12±0.43^b		218.36±15.42^b
FHD-H组		0.58±0.08^bc	2.18±0.26^bc		138.27±11.35^bc
PC组		0.65±0.09^bc	2.35±0.28^bc		145.38±12.17^bc
FHD-H+DMXAA组		1.52±0.15^d	4.86±0.47^d		248.65±16.28^d
F		155.527^＊＊	126.533^＊＊		157.966^＊＊
组别	GSH/ （μmol/g）		SOD/ （U/mg）	普鲁士蓝染色阳性面积比/%
Sham组	8.64±0.87		42.38±3.86	0.42±0.11
KOA组	2.35±0.31^a		15.28±1.94^a	8.76±0.94^a
FHD-L组	3.86±0.42^b		24.63±2.35^b	5.83±0.72^b
FHD-H组	6.72±0.58^bc		36.47±2.86^bc	1.85±0.34^bc
PC组	6.48±0.62^bc		35.26±2.74^bc	2.12±0.41^bc
FHD-H+DMXAA组	3.12±0.38^d		20.18±2.13^d	6.94±0.85^d
F	114.110^＊＊		90.650^＊＊	163.643^＊＊

基于STING/ACSL4通路探讨复元活血汤抑制膝骨关节炎大鼠软骨细胞铁死亡的作用机制

Mechanism of Fuyuan Huoxue decoction in inhibiting chondrocyte ferroptosis of rats with knee osteoarthritis via the STING/ACSL4 pathway

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组别	GPX4 阳性面积	SLC7A11 阳性面积	COX-2 阳性面积
Sham组	18.64±2.15	15.32±1.86	2.86±0.52
KOA组	3.28±0.64^a	4.15±0.73^a	21.48±2.35^a
FHD-L组	7.56±1.08^b	7.83±1.12^b	14.36±1.87^b
FHD-H组	15.38±1.64^bc	13.26±1.45^bc	5.27±0.94^bc
PC组	14.52±1.58^bc	12.48±1.52^bc	5.86±1.03^bc
FHD-H+DMXAA组	4.86±0.92^d	5.68±0.98^d	17.52±2.08^d
F	116.887^＊＊	69.423^＊＊	133.647^＊＊

组别	STING	ACSL4
Sham组	0.21±0.04	0.18±0.03
KOA组	0.86±0.09^a	0.92±0.10^a
FHD-L组	0.62±0.07^b	0.67±0.08^b
FHD-H组	0.32±0.05^bc	0.28±0.04^bc
PC组	0.35±0.06^bc	0.31±0.05^bc
FHD-H+DMXAA组	0.73±0.08^d	0.78±0.09^d
F	88.884^＊＊	114.061^＊＊

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