Efficacy mechanism of Yongjuxianfang oil in treating deep secondary-thickness burns by regulating MAPK pathway-mediated autophagy

YANG Meiyue; PAN Xinwen; LIAO Ruoyi; HUANG Xiangjun; YANG Hua

doi:10.12102/j.issn.2095-8668.2025.21.020

Abstract：Objective: To investigate the effect of Yingu Xianfang oil preparation on wound healing in deep second-degree burn mice and its molecular mechanism. Methods: Genes related to deep second-degree burn disease were retrieved, and active components and targets of the sovereign drug in Yingu Xianfang oil preparation were screened. After cross-screening, the interaction genes between Yingu Xianfang oil preparation target genes and deep second-degree burn target genes were selected. Twenty-four specific pathogen-free (SPF) male SD mice were used to create deep second-degree burn wounds on their backs using a hot iron block (75°C, 8 seconds), and were randomly divided into model group, inhibitor group, oil preparation group, and inhibitor + oil preparation group. Hematoxylin-eosin (HE) staining was used to observe the histomorphology and repair status of the wound tissue. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of vascular endothelial growth factor (VEGF) and fibroblast growth factor 2 (FGF-2), which are related to wound healing. Western blot (WB) was used to detect the levels of autophagy markers sequestosome 1 (P62), microtubule-associated protein 1A/1B light chain 3 (LC3)-I and LC3-II, and apoptosis marker cleaved-caspase 3. WB was also used to detect the expression of mitogen-activated protein kinase (MAPK) signaling pathway-related proteins extracellular signal-regulated kinase 1/2 (ERK1/2), c-Jun N-terminal kinase (JNK), and p38 mitogen-activated protein kinase (p38). Results: Starting from day 3 after the burn, the model group mice showed sloughing of the scab. After intervention with Yingu Xianfang oil preparation, the scab gradually softened, the edges lifted, and the base became flat with bleeding points after scab removal. From day 7 after the burn, there was a statistically significant difference in wound healing rate between the model group and the oil preparation group (P<0.01). Compared with the model group, the levels of wound healing factors in the oil preparation group increased (P<0.01), LC3-II was upregulated, while P62, LC3-I, cleaved-caspase 3, ERK1/2, JNK, and p38 protein expressions were all downregulated (P<0.01). After intervention with an autophagy inhibitor, LC3-II protein expression decreased, and the levels of VEGF and FGF-2 related to wound healing decreased. After Yingu Xianfang oil preparation intervention, they showed a trend of recovery, but were still lower than those in the oil preparation group alone (P<0.01). Conclusion: Yingu Xianfang oil preparation can promote the healing of deep second-degree burn wounds by inhibiting the MAPK pathway, inducing cell autophagy, inhibiting cell apoptosis, and increasing the levels of wound healing factors.