橙皮素通过抑制 p38 MAPK 信号通路减轻 T-2 毒素诱导的软骨细胞损伤。

Hesperetin Attenuates T-2 Toxin-Induced Chondrocyte Injury by Inhibiting the p38 MAPK Signaling Pathway.

机构信息

Institute for Endemic Fluorosis Control, Center for Endemic Disease Control, Chinese Center for Disease Control and Prevention, National Health Commission Key Laboratory of Etiology and Epidemiology, Harbin Medical University, Harbin 150081, China.

Heilongjiang Provincial Key Laboratory of Trace Elements and Human Health & Key Laboratory of Etiology and Epidemiology, Education Bureau of Heilongjiang Province, Harbin Medical University, Harbin 150081, China.

出版信息

Nutrients. 2024 Sep 14;16(18):3107. doi: 10.3390/nu16183107.

DOI:10.3390/nu16183107

PMID:39339707

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11434908/

Abstract

BACKGROUND

Hesperetin, a flavonoid derived from citrus fruits, exhibits potent antioxidant and anti-inflammatory activities and has been implicated in cartilage protection. However, its effectiveness against T-2 toxin-induced knee cartilage damage remains unclear.

METHODS

In this study, high-throughput sequencing analysis was employed to identify the key signaling pathways involved in T-2 toxin-induced articular cartilage damage in rats. Animal models were divided into the following groups: control, low-dose T-2 toxin, high-dose T-2 toxin, T-2 toxin + hesperetin, hesperetin, and vehicle. Pathological staining and immunohistochemistry were used to assess pathological changes, as well as the expression levels of the cartilage matrix-related proteins MMP13 and collagen II, along with the activation of the p38 MAPK signaling pathway. Additionally, primary rat chondrocytes were cultured to establish an in vitro model for investigating the underlying mechanism.

RESULTS

High-throughput sequencing analysis revealed the involvement of the MAPK signaling pathway in T-2 toxin-induced articular cartilage damage in rats. Hesperetin intervention in T-2 toxin-exposed rats attenuated pathological cartilage damage. Immunohistochemistry results demonstrated a significant reduction in collagen II protein expression in the high-dose T-2 toxin group ( < 0.01), accompanied by a significant increase in MMP13 protein expression ( < 0.01). In both the articular cartilage and the epiphyseal plate, the T-2 toxin + hesperetin group exhibited significantly higher collagen II protein expression than the high-dose T-2 toxin group ( < 0.05), along with significantly lower MMP13 protein expression ( < 0.05). Hesperetin inhibited the over-activation of the p38/MEF2C signaling axis induced by T-2 toxin in primary rat chondrocytes. Compared to the T-2 toxin group, the T-2 toxin + hesperetin group showed significantly reduced phosphorylation levels of p38 and protein expression levels of MEF2C ( < 0.001 or < 0.05). Moreover, the T-2 toxin + hesperetin group exhibited a significant decrease in MMP13 protein expression ( < 0.05) and a significant increase in collagen II protein expression ( < 0.01) compared to the T-2 toxin group.

CONCLUSIONS

T-2 toxin activates the p38 MAPK signaling pathway, causing knee cartilage damage in rats. Treatment with hesperetin inhibits the p38/MEF2C signaling axis, regulates collagen II and MMP13 protein expression, and reduces cartilage injury significantly.

摘要

背景

橙皮素是一种从柑橘类水果中提取的类黄酮，具有很强的抗氧化和抗炎活性，并与软骨保护有关。然而，其对 T-2 毒素诱导的膝软骨损伤的有效性尚不清楚。

方法

本研究采用高通量测序分析方法，鉴定 T-2 毒素诱导大鼠关节软骨损伤相关的关键信号通路。动物模型分为以下几组：对照组、低剂量 T-2 毒素组、高剂量 T-2 毒素组、T-2 毒素+橙皮素组、橙皮素组和载体组。采用病理染色和免疫组织化学法评估病理变化，以及软骨基质相关蛋白 MMP13 和胶原 II 的表达水平，以及 p38 MAPK 信号通路的激活情况。此外，还培养原代大鼠软骨细胞建立体外模型，探讨其潜在机制。

结果

高通量测序分析显示 MAPK 信号通路参与 T-2 毒素诱导的大鼠关节软骨损伤。橙皮素干预 T-2 毒素暴露大鼠可减轻病理软骨损伤。免疫组织化学结果显示，高剂量 T-2 毒素组（<0.01）胶原 II 蛋白表达显著降低，MMP13 蛋白表达显著升高（<0.01）。在关节软骨和骺板中，T-2 毒素+橙皮素组胶原 II 蛋白表达明显高于高剂量 T-2 毒素组（<0.05），MMP13 蛋白表达明显低于高剂量 T-2 毒素组（<0.05）。橙皮素抑制 T-2 毒素诱导的原代大鼠软骨细胞中 p38/MEF2C 信号轴的过度激活。与 T-2 毒素组相比，T-2 毒素+橙皮素组 p38 磷酸化水平和 MEF2C 蛋白表达水平均明显降低（<0.001 或<0.05）。此外，与 T-2 毒素组相比，T-2 毒素+橙皮素组 MMP13 蛋白表达明显降低（<0.05），胶原 II 蛋白表达明显升高（<0.01）。