目的 探討紅景天苷對(duì)高糖誘導(dǎo)的視網(wǎng)膜神經(jīng)節(jié)細(xì)胞（RGC）損傷及環(huán)磷酸腺苷/蛋白激酶A/cAMP反應(yīng)元件結(jié)合蛋白（cAMP/PKA/CREB）通路的影響。方法 將RGC-5細(xì)胞分為對(duì)照組、模型組、紅景天苷（0.25、0.5 mmol/L）組、紅景天苷＋Forskolin（0.5 mmol/L＋0.02 mmol/L）組。5-乙炔基-2’-脫氧尿嘧啶核苷（Edu）檢測各組細(xì)胞增殖情況；2,7-二氯熒光素二乙酸酯（DCFH-DA）熒光探針檢測活性氧（ROS）水平；酶聯(lián)免疫吸附試驗(yàn)（ELISA）檢測氧化應(yīng)激及cAMP水平；流式細(xì)胞儀檢測細(xì)胞凋亡；單丹磺酰尸胺（MDC）染色檢測自噬空泡形成；蛋白免疫印跡（Western blotting）檢測增殖、凋亡、自噬及cAMP/PKA/CREB通路相關(guān)蛋白表達(dá)。結(jié)果 與模型組比較，紅景天苷（0.25、0.5 mmol/L）組Edu陽性率、PCNA表達(dá)及超氧化物歧化酶（SOD）活性升高，活性氧（ROS）平均熒光強(qiáng)度、丙二醛（MDA）含量、細(xì)胞凋亡率、cleaved半胱氨酸蛋白酶-3（Caspase-3）、Caspase-3、自噬空泡綠色熒光強(qiáng)度、微管相關(guān)蛋白輕鏈3Ⅱ/LC3Ⅰ、Bcl-2同源結(jié)構(gòu)域蛋白抗體（Beclin-1）、PKA、p-CREB/CREB蛋白表達(dá)及cAMP水平顯著降低（P＜0.05）；與紅景天苷0.5 mmol/L組比較，紅景天苷＋Forskolin組Edu陽性率、PCNA表達(dá)及SOD活性降低，ROS平均熒光強(qiáng)度、MDA含量、細(xì)胞凋亡率、cleaved Caspase-3、Caspase-3、自噬空泡綠色熒光強(qiáng)度、LC3Ⅱ/LC3Ⅰ、Beclin-1、PKA、p-CREB/CREB蛋白表達(dá)及cAMP水平顯著升高（P＜0.05）。結(jié)論 紅景天苷可減輕高糖誘導(dǎo)的RGC-5細(xì)胞損傷，與抑制cAMP/PKA/CREB信號(hào)通路相關(guān)。

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

河北省衛(wèi)生健康委員會(huì)醫(yī)學(xué)科學(xué)研究課題計(jì)劃項(xiàng)目（20251433）