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目的观察补肾活血中药对体外晚期糖基化终末产物(AGEs)及缺氧条件下视网膜Müller细胞活力的影响。方法采用SD大鼠制备补肾活血中药含药血清和空白血清,以终浓度为1 mmol/L的连二亚硫酸钠制作细胞缺氧环境,以终浓度50 mg/L的AGEs作为低AGEs条件,终浓度100 mg/L的AGEs作为高AGEs条件,将体外纯化培养的P2代视网膜Müller细胞分为6组,分别置于加入空白血清、中药含药血清、空白血清+低AGEs+连二亚硫酸钠、空白血清+高AGEs+连二亚硫酸钠、中药含药血清+低AGEs+连二亚硫酸钠、中药含药血清+高AGEs+连二亚硫酸钠的DMEM培养基中,培养48 h后检测各组乳酸脱氢酶(LDH)漏出量,比较差异。结果 (1)低AGEs+缺氧组较正常对照组LDH值有升高趋势,但差异无统计学意义(P>0.05);高AGEs+缺氧组较正常对照组LDH值明显升高,其差异有统计学意义(P<0.05)。(2)正常中药干预组LDH数值与正常对照组接近,差异无统计学意义(P>0.05)。(3)两组不同剂量AGEs+缺氧中药干预组LDH漏出量均较对应的低、高AGEs+缺氧组明显降低,其差异有统计学意义(P<0.05)。结论 AGEs及缺氧条件可导致体外纯化培养的大鼠视网膜Müller细胞细胞膜稳定性下降,通透性增加,细胞活力降低,补肾活血中药含药血清可提高AGEs及缺氧条件下视网膜Müller细胞活力,这可能是补肾活血中药防治糖尿病性视网膜病变(DR)的机制之一。
Objective To observe the effects of Bushen Huoxue Chinese medicine on the activity of advanced glycation end products (AGEs) in vitro and the retinal Müller cells under hypoxia. Methods SD rat was used to prepare serum and serum of Bushen Huoxue Herbal Medicine. The hypoxia environment was made with sodium dithionite at a final concentration of 1 mmol / L. AGEs at a final concentration of 50 mg / L were used as the conditions of low AGEs, final concentration 100 mg / L AGEs as a condition of high AGEs, P2 cultured retinoblastoma cells were divided into 6 groups, which were placed in blank serum, Chinese medicine containing serum, blank serum + low AGEs + sodium dithionite, blank serum + High AGEs + sodium dithionite, Chinese medicine containing serum + low AGEs + sodium dithionite, Chinese medicine containing serum + high AGEs + sodium dithionite in DMEM medium for 48 h, the amount of lactate dehydrogenase (LDH) More difference. Results (1) The LDH values in low AGEs + hypoxia group were higher than those in normal control group, but the difference was not statistically significant (P> 0.05). The LDH value in high AGEs + hypoxia group was significantly higher than that in control group Statistical significance (P <0.05). (2) LDH values in normal TCM intervention group were close to those in normal control group, with no significant difference (P> 0.05). (3) The leakage of LDH in two groups of AGEs + hypoxia-treated group was lower than that of the corresponding group, and the difference was statistically significant (P <0.05). CONCLUSION AGEs and hypoxia can lead to the decrease of cell membrane stability, decrease of cell viability and decrease of cell viability of cultured retinal Müller cells cultured in vitro. The serum containing AGEs and retinal Müller cells under hypoxia can be increased by serum containing traditional Chinese medicine, This may be one of the mechanisms of Chinese medicine for tonifying kidney and promoting blood circulation to prevent and treat diabetic retinopathy (DR).