目的 探讨红景天苷对高糖诱导的视网膜神经节细胞（RGC）损伤及环磷酸腺苷/蛋白激酶A/cAMP反应元件结合蛋白（cAMP/PKA/CREB）通路的影响。方法 将RGC-5细胞分为对照组、模型组、红景天苷（0.25、0.5 mmol/L）组、红景天苷＋Forskolin（0.5 mmol/L＋0.02 mmol/L）组。5-乙炔基-2’-脱氧尿嘧啶核苷（Edu）检测各组细胞增殖情况；2,7-二氯荧光素二乙酸酯（DCFH-DA）荧光探针检测活性氧（ROS）水平；酶联免疫吸附试验（ELISA）检测氧化应激及cAMP水平；流式细胞仪检测细胞凋亡；单丹磺酰尸胺（MDC）染色检测自噬空泡形成；蛋白免疫印迹（Western blotting）检测增殖、凋亡、自噬及cAMP/PKA/CREB通路相关蛋白表达。结果 与模型组比较，红景天苷（0.25、0.5 mmol/L）组Edu阳性率、PCNA表达及超氧化物歧化酶（SOD）活性升高，活性氧（ROS）平均荧光强度、丙二醛（MDA）含量、细胞凋亡率、cleaved半胱氨酸蛋白酶-3（Caspase-3）、Caspase-3、自噬空泡绿色荧光强度、微管相关蛋白轻链3Ⅱ/LC3Ⅰ、Bcl-2同源结构域蛋白抗体（Beclin-1）、PKA、p-CREB/CREB蛋白表达及cAMP水平显著降低（P＜0.05）；与红景天苷0.5 mmol/L组比较，红景天苷＋Forskolin组Edu阳性率、PCNA表达及SOD活性降低，ROS平均荧光强度、MDA含量、细胞凋亡率、cleaved Caspase-3、Caspase-3、自噬空泡绿色荧光强度、LC3Ⅱ/LC3Ⅰ、Beclin-1、PKA、p-CREB/CREB蛋白表达及cAMP水平显著升高（P＜0.05）。结论 红景天苷可减轻高糖诱导的RGC-5细胞损伤，与抑制cAMP/PKA/CREB信号通路相关。

Objective To investigate the effects of salidroside on retinal ganglion cell (RGC) damage induced by high glucose and cAMP/PKA/CREB pathway. Methods RGC-5 cells were assigned into control group, model group, salidroside (0.25, 0.5 mmol/L) groups, and salidroside + Forskolin group. 5-Ethynyl-2'-deoxyuridine (Edu) was used to detect cell proliferation in each group. 2,7-dichlorofluorescein diacetate (DCFH-DA) fluorescent probe was used to detect reactive oxygen species (ROS). Enzyme linked immunosorbent assay (ELISA) was used to detect oxidative stress and cAMP levels. Flow cytometry was used to detect cell apoptosis. Monodansylcadaverine (MDC) staining was used to detect autophagic vacuolar formation. Western blotting was used to detect the expression of proteins related to proliferation, apoptosis, autophagy, and the cAMP/PKA/CREB pathway. Results Compared with model group, the Edu positive rate, PCNA expression, and SOD activity in salidroside (0.25, 0.5 mmol/L) groups were increased, ROS mean fluorescence intensity, MDA content, apoptosis rate, cleaved Caspase-3, Caspase-3, autophagy vacuolar green fluorescence intensity, LC3Ⅱ/LC3Ⅰ, Beclin-1, PKA, p-CREB/CREB protein, and cAMP level were significantly decreased (P < 0.05). Compared with salidroside 0.5 mmol /L group, the Edu positive rate, PCNA expression, and SOD activity of salidroside +Forskolin group were decreased, ROS mean fluorescence intensity, MDA content, apoptosis rate, cleaved Caspase-3, Caspase-3, autophagy vacuolar green fluorescence intensity, LC3Ⅱ/LC3Ⅰ, Beclin-1, PKA, p-CREB /CREB protein expression, and cAMP level were significantly increased (P < 0.05). Conclusion Salidroside can alleviate high glucose induced RGC-5 injury, and it is associated with inhibition of cAMP/PKA/CREB signaling pathway.

R285.5

河北省卫生健康委员会医学科学研究课题计划项目（20251433）