为研究哈蟆油（Oviductus Ranae，OR）对海马神经元细胞保护作用机制， 该研究通过皮质酮（Corticosterone，CORT）诱导HT-22细胞损伤模型探究哈蟆油酶解物对小鼠海马神经元细胞（HT-22）细胞损伤模型的保护作用及其作用机制。采用CORT与HT-22细胞共同培养建立HT-22细胞损伤模型，给予哈蟆油酶解物再次孵育，将ERK信号通路抑制剂PD98059加入HT-22细胞中。采用MTT法、Hochest 33258染色、流式细胞术、Western blot技术检测HT-22细胞凋亡情况和相关蛋白表达以及MAPK/ERK信号通路蛋白表达。结果显示高水平CORT能够诱导HT-22细胞损伤，哈蟆油酶解物能够使CORT诱导的HT-22细胞活力提升62.5%~87.5%，凋亡比例降低50%~70%。哈蟆油酶解物能够上调BCL-2、P-ERK1/2蛋白表达，同时下调BAX、Caspase-3、Caspase-9的蛋白表达。上述结果表明，高水平CORT是通过抑制MAPK/ERK信号通路促进凋亡的发生从而诱导HT-22细胞损伤，哈蟆油酶解物能够通过激活MAPK/ERK信号通路调节BLC-2、BAX、Caspase-3、Caspase-9凋亡相关蛋白的表达进而抑制凋亡的发生减轻神经元损伤，该作用机制为哈蟆油发挥抗抑郁作用的关键途径之一。

To examine the protective mechanism of oviductus ranae (OR) on hippocampal neuronal cells, this study explored the protective effect and mechanism of OR hydrolysate in a corticosterone (CORT)-induced mouse hippocampal neuron (HT-22) model of cell injury. HT-22 cells were co-cultured with CORT to establish an injury model, then further incubated with OR hydrolysate, followed by the addition of the ERK signaling pathway inhibitor PD98059. MTT assay, Hoechst 33258 staining, flow cytometry and Western blot were employed to assess cell apoptosis, protein expression, and MAPK/ERK signaling pathway activity. Results demonstrated that elevated levels of CORT induced damage in HT-22 cells, while OR hydrolysate enhanced cell viability by 62.5%~87.5%, and reduced apoptosis rates by 50%~70%. Moreover, OR hydrolysate upregulated BCL-2 and P-ERK1/2 protein expression, and downregulated BAX, Caspase-3 and Caspase-9 protein expressions. These findings indicate that high CORT levels promote apoptosis by inhibiting the MAPK/ERK signaling pathway, leading to HT-22 cell damage. OR hydrolysate regulates the expression of apoptosis-related proteins, (BLC-2, BAX, Caspase-3 and Caspase-9) by activating the MAPK/ERK signaling pathway, thereby inhibiting apoptosis and reducing neuronal damage. This mechanism represents a key pathway through which OR exerts anti-depressive effects.

吉林省科技厅项目（20220505038ZP ；20210401067YY）