【摘要】 目的观察在晚期糖基化终末产物(AGEs)条件下,补肾活血方含药血清对大鼠视网膜Müller细胞的缺氧诱导因子-1α(HIF-1α)缺氧信号通路的影响。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、空白血清+高AGEs、中药含药血清+低AGEs、中药含药血清+高AGEs的DMEM培养基中,培养48 h后酶联免疫吸附法(ELISA)检测各组细胞外液中HIF-1α、血管内皮生长因子(VEGF)含量,逆转录-聚合酶链法(RT-PCR)测定各组Müller细胞HIF-1αm RNA、VEGFm RNA表达量,比较差异。结果 1.低AGEs组较正常对照组HIF-1α值有升高趋势,但差异无统计学意义(P>0.05),HIF-1αm RNA值明显升高,其差异有统计学意义(P<0.05),高AGEs组较正常对照组HIF-1α、HIF-1αm RNA值明显升高,其差异有统计学意义(P<0.05);低、高AGEs组较正常对照组VEGF、VEGFm RNA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HIF-1α、VEGF、HIF-1αm RNA、VEGFm RNA值接近正常对照组,差异无统计学意义(P>0.05)。3.低、高AGEs中药干预组HIF-1α、VEGF、HIF-1αm RNA、VEGFm RNA值均较对应的低、高AGEs组明显降低,其差异有统计学意义(P<0.05)。结论补肾活血方含药血清可抑制视网膜Müller细胞在AGEs条件下HIF-1α介导缺氧信号通路的高表达,这可能是补肾活血方防治DR的机制之一。更多还原

【Abstract】 OBJECTIVE To investigate the influence of serum with Bushen Huoxue(BSHX) decoction on signal pathway mediated by hypoxia-inducible factors 1α(HIF-1α) of retinal Müller cells(RMCs) cultured in advanced glycation end products(AGEs) condition. METHODS The serum containing Chinese medicine BSHX decoction and blank serum was prepared using SD rats. The concentration of AGEs in 50 mg/L was set as low AGEs condition, and100 mg/L as high AGEs condition. The purified P2 retinal Müller cells of newborn and 7 day-rats were cultured by modified enzyme-digestion method. Purified RMCs were divided into 6 groups, all of them were cultured in Dulbecco’s Modified Eagle’s medium(DMEM) and then were added in blank serum, medicated serum, blank serum with low AGEs, blank serum with high AGEs, medicated serum with low AGEs, medicated serum with high AGEs under hy-poxic condition respectively. After 48 hrs, the HIF-1α, vascular endothelial growth factor(VEGF) expression in extracellular fluid were detected by enzyme-linked immunosorbent assay(ELISA), while expression of m RNA of HIF-1αand VEGF in Müller cells were tested by reverse transcription polymerase chain reaction(RT-PCR) and results of different groups were compared. RESULTS The expression of HIF-1α in the low AGEs group was higher than normal control group, but the difference was of no statistic difference(Ρ >0.05), but the increase of HIF-1α m RNA made a significant difference(Ρ<0.05);the expression of HIF-1α and its m RNA in the high AGEs was significantly higher in contrast to normal control group(Ρ<0.05). The expressions of VEGF and its m RNA in the low AGEs and high AGEs group were both significantly higher than normal control group(Ρ<0.05). While BSHX medicine-containing serum had no significant effect on the expression of HIF-1α, VEGF and their m RNA compared with normal control conditions(Ρ>0.05). Both the expression of HIF-1α, VEGF and their m RNA decreased more dramatically by BSHX decoction intervention under different AGEs conditions in contrast to corresponding AGEs concentration without intervention(Ρ <0.05).CONCLUSIONS The BSHX serum inhibited the expression of signal pathway mediated by HIF-1α in purified RMCs under AGEs condition. Accordingly, it might be part of the mechanism explaining effects of BSHX on diabetic retinopathy.更多还原