目的:探讨黄连素对高糖培养大鼠离体视网膜Müller细胞的影响及其作用机制。方法:将体外培养的Sprague Dawley(SD)大鼠视网膜Müller细胞分为正常对照组、高糖组、高糖＋10 μmol/L黄连素组和高糖＋25 μmol/L黄连素组，正常对照组和高糖组细胞分别采用5 mmol/L葡萄糖和25 mmol/L葡萄糖培养，后2个组分别采用25 mmol/L葡萄糖＋相应浓度黄连素处理，培养后72 h采用流式细胞仪检测细胞凋亡情况，采用酶联免疫吸附测定(ELISA)法及实时荧光定量PCR法分别检测细胞培养液上清中炎性因子肿瘤坏死因子(TNF-α)、白细胞介素-8(IL-8)和环氧合酶2(COX-2)，L-谷氨酸-L-天冬氨酸转运体(GLAST)及相关蛋白的表达情况，采用Western blot法检测细胞质中GLAST、镁离子依赖的蛋白磷酸酶1A(PPM1A)、核因子-κB(NF-κB)和cleaved caspase-3及细胞核中NF-κB蛋白的表达情况。结果:正常对照组、高糖组、高糖＋10 μmol/L黄连素组和高糖＋25 μmol/L黄连素组细胞凋亡率分别为(1.37±0.21)%、(17.67±1.17)%、(10.60±0.17)%和(5.57±0.35)%，总体比较差异有统计学意义(n F＝375.97，n P<0.01)，其中高糖组细胞凋亡率较正常对照组明显升高，高糖＋10 μmol/L黄连素组和高糖＋25 μmol/L黄连素组细胞凋亡率较高糖组明显降低，高糖＋25 μmol/L黄连素组较高糖＋10 μmol/L黄连素组细胞凋亡率明显降低，差异均有统计学意义(均n P<0.01)。ELISA检测结果显示，各组细胞中TNF-α、IL-8、COX-2质量浓度总体比较差异均有统计学意义(n F＝28.36、35.88、41.59，均n P<0.01)，其中高糖组细胞中TNF-α、IL-8和COX-2质量浓度均明显高于正常对照组，高糖＋10 μmol/L黄连素组和高糖＋25 μmol/L黄连素组细胞中TNF-α和COX-2质量浓度明显低于高糖组，高糖＋25 μmol/L黄连素组细胞中TNF-α和IL-8浓度明显低于高糖＋10 μmol/L黄连素组，差异均有统计学意义(均n P<0.05)。实时荧光定量PCR结果显示，各组间Müller细胞中GLAST mRNA的相对表达量总体比较差异有统计学意义(n F＝268.60，n P<0.01)，其中高糖组GLAST mRNA的相对表达量明显低于正常对照组、高糖＋10 μmol/L黄连素组和高糖＋25 μmol/L黄连素组，差异均有统计学意义(均n P<0.05)。Western blot分析结果显示，各组Müller细胞质中GLAST、PPM1A、cleaved caspase-3和NF-κB蛋白相对表达量以及细胞核中NF-κB蛋白相对表达量总体比较差异均有统计学意义(n F＝135.20、156.98、80.96、128.07、47.36，均n P<0.01)，其中高糖组Müller细胞质中GLAST、PPM1A和NF-κB蛋白的相对表达量明显低于正常对照组、高糖＋10 μmol/L黄连素组和高糖＋25 μmol/L黄连素组，细胞质中cleaved caspase-3和细胞核中NF-κB蛋白的相对表达量明显高于正常对照组、高糖＋10 μmol/L黄连素组和高糖＋25 μmol/L黄连素组，差异均有统计学意义(均n P<0.05)。n 结论:高糖可诱导SD大鼠视网膜Müller细胞发生凋亡及炎症反应，而黄连素能有效抑制高糖诱发的细胞凋亡及炎症反应，其可能是通过抑制NF-κB的核易位和转录活性，从而抑制炎性因子的表达来发挥保护作用。“,”Objective:To investigate the effects of berberine on Sprague Dawley (SD) rat retinal Müller cells cultured by high concentration glucose.Methods:The cultured SD rat retinal Müller cells were divided into normal-glucose group, high-glucose group, high-glucose+ 10 μmol/L berberine group and high-glucose+ 25 μmol/L berberine group, and the cells were cultured in 5 mmol/L glucose, 25 mmol/L glucose, 25 mmol/L glucose+ 10 μmol/L berberine, and 25 mmol/L glucose+ 25 μmol/L berberine, respectively.After 72 hours cultured, cell apoptosis rate was detected by flow cytometry; the expressions levels of tumor necrosis factor-α (TNF-α), interleukin-8 (IL-8) and cyclooxygenase-2 (COX-2) in the culture supernatant were detected by enzyme linked immunosorbent assay (ELISA); the L-glutamate-L-aspartate transporter (GLAST) and the related protein expression levels were detected by real-time fluorescence quantitative PCR; the expressions levels of GLAST, protein phosphatase magnesium-dependent 1A (PPM1A), nuclear factor-κB (NF-κB) and cleaved caspase-3 in the cytoplasm, and the expression level of NF-κB protein in the nucleus were detected by Western blot.Results:The cell apoptosis rate was (1.37±0.21)%, (17.67±1.17)%, (10.60±0.17)% and (5.57±0.35)% in the normal-glucose group, high-glucose group, high-glucose+ 10 μmol/L berberine group and high-glucose+ 25 μmol/L berberine group, respectively, and the overall comparative difference was statistically significant ( n F=375.97, n P<0.01). The cell apoptosis rates in the high-glucose group was increased in comparison with those in the normal-glucose group (n P<0.01). The cell apoptosis rates in the high-glucose+ 10 μmol/L berberine group and high-glucose+ 25 μmol/L berberine group were significantly reduced in comparison with that in the high-glucose group (both atn P<0.01). And the cell apoptosis rate in the high-glucose+ 25 μmol/L berberine group was lower than that in the high-glucose+ 10 μmol/L berberine group (n P<0.01). ELISA results revealed that the overall comparative differences of the concentrations of TNF-α, IL-8 and COX-2 among the four groups were statistically significant (n F=28.36, 35.88, 41.59; all at n P<0.01). The concentrations of TNF-α, IL-8 and COX-2 in the high-glucose group were significantly higher than those in the normal-glucose group (n P<0.01). Compared with the high-glucose group, the concentrations of TNF-α and COX-2 were significantly decreased in the high-glucose+ 10 μmol/L berberine group and high-glucose+ 25 μmol/L berberine group (both atn P<0.05). The concentrations of TNF-α and IL-8 in the high-glucose+ 25 μmol/L berberine group were lower than those in the high-glucose+ 10 μmol/L berberine group (both atn P<0.05). Real-time fluorescence quantitative PCR showed that the relative expression levels of GLAST mRNA in Müller cells among the four groups were statistically significant (n F=268.60, n P<0.01). Compared with the normal-glucose group, the relative expression level of GLAST mRNA was significantly decreased in the high-glucose group (n P<0.01). In the high-glucose+ 10 μmol/L berberine group and high-glucose+ 25 μmol/L berberine group, the relative expression level of GLAST mRNA was significantly elevated compared with the high-glucose group (both atn P<0.05). Western blot analysis results showed that the overall comparative differences of the GLAST, PPM1A, cleaved caspase-3, NF-κB in cytoplasm and NF-κB in nucleus among the four groups were statistically significant (n F=135.20, 156.98, 80.96, 128.07, 47.36; all at n P<0.01). The relative expression levels of GLAST, PPM1A and NF-κB protein in cytoplasm in the high-glucose group were significantly lower than those in the normal-glucose group, high-glucose+ 10 μmol/L berberine group and high-glucose+ 25 μmol/L berberine group (all atn P<0.05). The relative expression levels of cleaved caspase-3 and NF-κB protein in nucleus in the high-glucose group were significantly higher than those in the normal-glucose group, high-glucose+ 10 μmol/L berberine group and high-glucose+ 25 μmol/L berberine group (all atn P<0.05).n Conclusions:High-concentration glucose can induce cell apoptosis and inflammatory response of SD rats retinal Müller cells n in vitro.However, berberine can inhibit cell apoptosis and inflammatory response induced by high-concentration glucose via suppressing NF-κB translocation and transcription activity, and thereby inhibiting the expression of inflammatory cytokines.n