丹参脂溶性成分对AGEs条件下视网膜Müller细胞HIF-1及谷氨酰胺合成酶表达的影响

The effect of the liposoluble components of Salvia on rat retinal Müller cells HIF-1 and GS in AGEs condition

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 effect of Salvia lipophilic extract on lactate dehydrogenase （LDH） leakage and the expression of HIF-1 and GS in vitro retinal MOiler cells under advanced glycation end products（AGEs） condition. Method： 5 μg,/ml liposoluble components of Salvia and 5 μg/rnl Tanshinone IIA were used to intervene rat retinal Mtiller cells, and detected the LDH leakage and the expression of HIF-1 and GS in Muller cells were cultured in normal, high and low concentration （ 150 μg/ml, 75 μg/ml） of AGEs condition by the enzyme-linked im- munosorbent assay （ELISA）. Result： Compared with the 48 h group, 24 h group of LDH leakage was decreased, and 72 h group of LDH leakage was significantly increased （ P 〈 0.05 ） under normal and AGEs condition. The liposoluble components of Salvia and Tanshinone IIA LDH leakage were decreased compared with the blank group; Under AGEs condition, the expression of HIF-1 were significantly higher than normal condition （ P 〈 0.05） , and GS were significantly lower than normal conditions （ P 〈 0.05）. Compared with the blank group, The li- posoluble components of Salvia and Tanshinone IIA could significantly decreased the expression levels of HIF-1 （P 〈 O. 05 ） and increase the expression of GS in Miller ceils （ P 〈 0.05 ）. Conclusion： Each group Mtiller cells vitality was reduced after rising, at 48 h Muller cells vi- tality was the highest, so choose 48 h as optimal intervention time. The expression of HIF-1 was increased and the expression of GS was in- hibiticd in rat retinal Mailer cells by AGEs, the liposoluble components of Salvia and Tanshinone IIA could inhibit the expression of HIF-1 and enhance the expression of GS, thus reducing the vascular endothelial growth factor （VEGF） generation and the concentration of glutamate （Glu）, and play an important role for neuro protection which provides some experimental basis to further study the mechanism of the treat- ment of DR with hposoluble components of Salvia .