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目的应用中药血清药理学方法探讨补肾活血中药复方对体外共同培养的视网膜神经节细胞和Müller细胞谷氨酸(glutamate,Glu)代谢的影响。方法氨基酸分析仪检测共同培养细胞外液中Glu释放量,[3H]标记的D,L-Glu摄入量判断共同培养细胞的Glu摄取功能测定中药血清干预高糖和(或)缺氧状态下体外共同培养细胞外液中Glu的释放氉量和摄入量。结果在48 h时段,缺氧组及高糖缺氧组共同培养细胞的Glu释放量均比正常对照组明显增加(均为P<0.05),而高糖组Glu释放量比正常对照组明显减少(P<0.05);补肾活血中药血清能明显降低正常条件下(24 h、48 h、72 h)、缺氧及高糖缺氧条件下(48 h、72 h)共同培养细胞的释放量(均为P<0.05)。在24 h、48 h、72 h时段缺氧组及高糖缺氧组共同培养细胞的Glu摄入量均比正常对照组明显降低(均为P<0.05);高糖组共同培养细胞的Glu摄取功能在24 h和72 h时段均比正常对照组增强(均为P<0.05),但48 h时段Glu摄入比正常对照组降低(P<0.05);补肾活血中药血清能明显提高正常及缺氧条件下24 h、48 h时段、高糖条件下48 h时段以及高糖和(或)缺氧条件下72 h时段共同培养细胞的Glu摄入量(均为P<0.05)。结论高糖和(或)缺氧能够引起体外共同培养的视网膜神经节细胞和视网膜Müller细胞的Glu代谢障碍,这可能是补肾活血法对高糖和(或)缺氧状态下视网膜神经节细胞的保护作用机制之一。
Objective To study the effect of Chinese herbs for replenishing kidney and activating blood circulation on the metabolism of glutamate (Glu) in cultured retinal ganglion cells and Müller cells cultured in vitro by using the method of traditional Chinese medicine serum pharmacology. Methods Amino acid analyzer was used to determine Glu release in co-cultured extracellular fluid and determination of Glu uptake in cocultured cells by [3H] labeled D, L-Glu intake. Serum levels of Chinese herbal medicine were determined by high glucose and / or hypoxia In vitro co-culture of extracellular fluid in the Glu release quantity and intake. Results During 48 h, the Glu release in cocultured cells in hypoxia group and hypoxia-hypoxia group were significantly higher than those in normal control group (all P <0.05), while Glu release in high glucose group was significantly lower than that in normal control group (P <0.05). The serum of Bushen Huoxue Decoction could significantly reduce the co-cultured cells release under normal conditions (24 h, 48 h, 72 h), hypoxia and hypoglycemia (48 h, 72 h) All P <0.05). Compared with the normal control group, the Glu intake of the cocultured cells in the hypoxia group and the high glucose hypoxia group were significantly lower than those in the normal control group (all P <0.05) at 24 h, 48 h and 72 h, Compared with the normal control group, the intake of Glu (P <0.05) increased at 24 h and 72 h (P <0.05), but the Glu intake at 48 h was lower than that of the normal control group (P <0.05) During 24 h, 48 h, 48 h hypoglycemic and 48 h hypoglycemic conditions, Glu intake of cocultured cells in high glucose and / or hypoxia for 72 h (all P <0.05). Conclusion High glucose and / or hypoxia can cause Glu metabolic disorder in cultured retinal ganglion cells and retinal Müller cells in vitro. This may be due to the effect of tonifying kidney and promoting blood circulation on retinal ganglion cells in high glucose and / or hypoxia One of the mechanisms of protection.