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目的:研究丹参脂溶性成分对体外纯化培养的大鼠视网膜Müller细胞在糖基化终末产物(advanced glycation end products,AGEs)条件下乳酸脱氢酶(lactate dehydrogenase,LDH)的漏出量、低氧诱导因子(hypoxia-inducible factor-1,HIF-1)及谷氨酰胺合成酶表达的影响。方法:以5μg/ml的丹参脂溶性成分及5μg/ml丹参酮IIA分别干预视网膜Müller细胞,以ELISA法分别测定在正常及高、低浓度AGEs(150μg/ml、75μg/ml)条件下LDH漏出量以及HIF-1、谷氨酰胺合成酶的表达情况。结果:在正常及AGEs条件下,各组Müller细胞LDH的漏出量48 h较24 h明显减少,72 h较48 h明显增多,丹参脂溶性成分及丹参酮IIA组LDH漏出量较空白对照组小;在AGEs条件下,空白对照组HIF-1表达量较正常条件下显著增高;谷氨酰胺合成酶的表达量显著低于正常条件。丹参酮IIA、丹参脂溶性成分组与空白对照组相比,均能显著抑制HIF-1表达,增加谷氨酰胺合成酶的表达。结论:各实验组Müller细胞活力均呈现先升高、后降低,在48 h时Müller细胞膜活力最高。因此选择48 h作为最佳干预时间。AGEs能明显提高大鼠视网膜Müller细胞HIF-1的表达,抑制谷氨酰胺合成酶的表达。丹参脂溶性成分及丹参酮IIA均能抑制AGEs条件下大鼠视网膜Müller细胞HIF-1的表达,增强谷氨酰胺合成酶的表达,从而降低血管内皮生长因子(vascular endothelial growth factor,VEGF)的生成量以及谷氨酸(Glutamate,Glu)的浓度,研究结果为进一步探讨丹参脂溶性成分治疗糖尿病性视网膜病变作用机制提供了一定的实验依据。
OBJECTIVE: To study the effects of lipophilic components of Radix Salviae Miltiorrhizae on the leakage of lactate dehydrogenase (LDH) in cultured rat retinal Müller cells under advanced glycation end products (AGEs) Effects of hypoxia-inducible factor-1 (HIF-1) and glutamine synthetase METHODS: Retinal Müller cells were treated with 5μg / ml Salvia miltiorrhiza fat-soluble fraction and 5μg / ml tanshinone IIA respectively. The amount of LDH leakage in normal and high and low AGEs (150μg / ml, 75μg / ml) And HIF-1, glutamine synthetase expression. Results: Under normal and AGEs conditions, the leakage of LDH in Müller cells in each group decreased 48 h compared with that in 24 h, but increased significantly in 72 h and 48 h compared with that in control group. The leakage of LDH in lipid extracts of Danshen and Tanshinone IIA groups was smaller than that in control group. Under the conditions of AGEs, the expression of HIF-1 in the blank control group was significantly higher than that in the normal conditions; the expression of glutamine synthetase was significantly lower than the normal conditions. Tanshinone IIA, Salvia miltiorrhiza lipid-soluble components compared with the blank control group, were significantly inhibited HIF-1 expression, increased glutamine synthetase expression. Conclusion: The Müller cells in each experimental group showed the first increase in activity and the second decrease in Müller cells. The Müller cell membrane activity was the highest at 48 h. So choose 48 h as the best intervention time. AGEs can significantly improve the expression of HIF-1 in retina Müller cells and inhibit the expression of glutamine synthetase. Salvia miltiorrhiza lipid-soluble components and tanshinone IIA can inhibit the expression of HIF-1, enhance the expression of glutamine synthetase and decrease the production of vascular endothelial growth factor (VEGF) in retina Müller cells under AGEs As well as the concentration of glutamate (Glutamate, Glu), the results provide some experimental evidence for further exploring the mechanism of action of lipid-soluble components of Salvia miltiorrhiza in the treatment of diabetic retinopathy.