旱莲苷A通过抑制MAPK/P53通路介导的抗炎与抗凋亡作用改善干眼症

Ecliptasaponin A ameliorates dry eye via anti-inflammatory and anti-apoptotic effects through inhibition of the MAPK/P53 pathway

摘要

目的 探讨旱莲苷A (EA) 对干眼症 (DE) 模型小鼠的药效作用和初步机制研究。方法 建立DE的小鼠模型, 通过荧光素钠染色、酚红棉线试验和HE染色验证EA对DE的药效作用。ELISA法测定小鼠白细胞介素1β、白细胞介素6和肿瘤坏死因子α等指标。此外, 通过活性氧、F4/80和TUNEL免疫荧光染色评估EA对小鼠DE后角膜炎症和细胞凋亡的改善作用。Western blotting实验检测丝裂原活化蛋白激酶 (MAPK) 通路、P53通路相关蛋白磷酸化水平。结果 对照组各项检测指标均未见明显异常。与模型组相比, EA各剂量治疗组泪液分泌量增加 (P<0.01)、角膜损伤评分降低 (P<0.01)、角膜病理损伤改善 (P<0.01)、炎性因子表达水平降低 (P<0.01) 且p-TAK1、TAK1、p-P38、P38、p-P53和P53的蛋白水平明显降低 (P<0.01)。结论 EA可能通过抑制MAPK通路、P53通路减轻炎症和凋亡反应, 在DE中发挥保护作用, 为EA的临床应用提供实验依据。

Abstract

Objective To explore the pharmacological effect and preliminary mechanism of ecliptasaponin A (EA) on dry eye (DE) model mice. Methods A mouse model of DE was established, and the pharmacological effect of EA on DE was verified through fluorescein sodium staining, phenol red thread test and HE staining. ELISA was used to measure indicators such as interleukin-1β, interleukin-6, and tumor necrosis factor-α in mice. In addition, the improvement effect of EA on corneal inflammation and cell apoptosis after DE in mice was evaluated through reactive oxygen species, F4/80 and TUNEL immunofluorescence staining. Western blotting was used to detect the phosphorylation levels of proteins related to mitogen-activated protein kinase (MAPK) pathway and P53 pathway. Results The various detection indicators in the control group showed no significant abnormalities. Compared with the DE model group, the treatment groups with different doses of EA had increased tear secretion (P<0.01), decreased corneal injury score (P<0.01), improved corneal pathological damage (P<0.01), decreased expression levels of inflammatory factors (P<0.01), and significantly lower protein levels of p-TAK1, TAK1, p-P38, P38, p-P53, and P53 (P<0.01). Conclusion EA may exert a protective effect in DE by inhibiting the MAPK pathway and the P53 pathway, thereby alleviating inflammatory and apoptotic responses. This provides experimental evidence for the clinical application of EA.

关键词

干眼症 / 旱莲苷A / 抗炎 / 抗凋亡 / MAPK / P53 / P38 / TAK1

Key words

dry eye / ecliptasaponin A / anti-inflammatory / anti-apoptotic / MAPK / P53 / P38 / TAK1

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基金资助

国家自然科学基金青年科学基金(82405134)