人参皂苷对糖基化终末产物条件下视网膜Müller细胞活力的影响

Effect of Ginsenoside on Retinal Müller Cell Cultured in Advanced Glycation End Products Conditions

摘要:

目的：探讨人参皂苷对糖基化终末产物（advanced glycation end products，AGEs）条件下视网膜Müller细胞活力的影响。方法：以改良酶消化法培养SD大鼠视网膜Müller细胞，将纯化培养的视网膜Müller细胞分别置于低剂量AGEs（终质量浓度为75 mg·L^-1）及高剂量AGEs（终质量浓度为150 mg·L^-1）下进行培养，并分别以人参皂苷Rg₁、人参皂苷Re、人参皂苷Rb₁及人参皂苷提取物干预。在干预24，48，72 h后以酶联免疫吸附法测定各组细胞外液中乳酸脱氢酶（lactate dehydrogenase，LDH）的漏出量，以推测Müller细胞活力。结果：低AGEs组24，48 h的LDH漏出量较正常对照组均无明显差异，但72 h的LDH漏出量较正常对照组增多（P<0.05）；高AGEs组各时段LDH漏出量均较正常对照组以及低AGEs组增多（P<0.05）；人参皂苷Rg₁，Re，Rb₁及人参皂苷提取物均能减少低AGEs组48，72 h的LDH漏出量（P<0.05），减少高AGEs组各时段的LDH漏出量（P<0.05）。结论：AGEs能明显增加视网膜Müller细胞膜的通透性，降低细胞活力，并且与干预的时间、浓度有关；人参皂苷Rg₁，Re，Rb₁及人参皂苷提取物能降低本实验中AGEs条件下Müller细胞膜的通透性、提高Müller细胞膜稳定性，增强细胞活力，尤其以人参皂苷提取物效果显著。

Abstract:

Objective: To explore the effect of ginsenoside on retinal Müller cell activity under the conditions of advanced glycation end products (AGEs). Method: Modified enzyme-digesting method was used to culture SD rat retinal Müller cells, retinal Müller cells were then cultured in low (terminal concentration:75 mg·L^-1) dose of AGEs conditions and high (terminal concentration:150 mg·L^-1) dose of AGEs conditions, ginsenoside Rg₁, ginsenoside Re, ginsenoside Rb₁ and ginsenoside extractions were added to culture medium respectively. At 24, 48, 72 hours after incubation, the lactate dehydrogenase (LDH) content of extracellular fluid were detected by the enzyme-linked immunosorbent assay (ELISA) to determine Müller cell activity. Result: Compared with normal group the LDH leakage was significantly increased in the low dose of AGEs group at 72 h (P<0.05), whereas there was no difference at 24 h and 48 h;compared with normal group and low dose of AGEs group the LDH leakage were significantly increased in the high dose of AGEs group at each period (P<0.05) ;ginsenoside Rg₁, ginsenoside Re, ginsenoside Rb₁ and ginsenoside extractions could significantly decrease the LDH leakage in the low dose of AGEs group at 48 h and 72 h, and in the high dose of AGEs group at each period (P<0.05). Conclusion: AGEs caused a significant increase of the membrane permeability of retinal Müller cell, decreased the activity of retinal Müller cell, which was related to the time and concentration of intervention;ginsenoside Rg₁, ginsenoside Re, ginsenoside Rb₁ and ginsenoside extractions could decrease the membrane permeability, could stabilize cells' membrane and enhance the activity of retinal Müller cell in AGES conditions, in particular, the effect of ginsenoside extractions is significant.