补肾活血中药血清对高糖状态下纯化培养的视网膜神经节细胞活力的影响

Effect of Serum Contained Chinese Drugs for Nourishing Shen and Activating Blood on Activity of Purified Retinal Ganglion Cells Cultured in High Glucose Medium

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中文关键词: 高糖糖波动视网膜神经节细胞乳酸脱氢酶补肾活血

英文关键词:high glucose glucose fluctuation retinal ganglion cell lactate dehydrogenase nourishing Shen and activating blood

基金项目:国家自然科学基金资助项目(No.30572407);四川省科技厅应用基础项目(No.008JY0085);四川省教育厅重点项目(No.07EA023)

作者	单位
马荣	成都中医药大学研究生院
谢学军	成都中医药大学眼科教研室
万李	成都中医药大学研究生院
马殿伟	成都中医药大学研究生院

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中文摘要:

目的探讨补肾活血中药血清对体外高糖状态下纯化培养的视网膜神经节细胞活力的影响。方法体外纯化SD大鼠视网膜神经节细胞(retinal ganglion cells,RGCs),模拟稳定高糖(50mmol/L)及糖波动环境进行培养,以补肾活血中药血清进行干预,检测RGCs乳酸脱氢酶(lactate dehydrogenase,LDH)漏出量,以推测RGCs活力。结果RGCs的LDH漏出量(U/L)稳定高糖组24h(1349.17±215.50)、48h(1220.24±124.53)及72h(1982.14±219.03)均较正常对照组(1628.10±122.10、1484.13±127.55及2155.75±140.44)降低(P<0.05);而糖波动组的LDH漏出量在72h(2299.60±88.35)较正常对照组增加(P<0.05),且糖波动组各时段的LDH漏出量均较稳定高糖组增加(P<0.05);稳定高糖中药干预组的LDH漏出量在72h(1797.62±146.40)时较稳定高糖组减少(P<0.05);糖波动中药干预组的LDH漏出量在48h(1259.92±87.74)和72h(1940.40±155.47)时均较糖波动组减少(P<0.05)。结论糖波动能明显降低RGCs细胞膜的稳定性,增加细胞膜通透性,降低细胞活力;补肾活血中药血清能降低本实验中稳定高糖及糖波动条件下RGCs的细胞膜通透性,提高细胞膜稳定性,增强其细胞活力,这可能是补肾活血中药复方防治糖尿病视网膜病变(diabetic retinopathy,DRP)的药物干预途径之一。

英文摘要:

Objective To investigate the effect of serum contained Chinese drugs for nourishing Shen and activating blood(S-NSAB)on activity of purified retinal ganglion cells(RGCs)cultured in high glucose medium.Methods Purified RGCs of SD rats were cultured in simulative stable high glucose(50 mmol/L)condition(SHG)and fluctuated glucose condition(FGC)separately,they were intervened with S-NSAB,and the lactate dehydrogenase(LDH)leakage was detected by spectrophotometer for estimating the activity of RGCs.Results LDH leakage(U/L)in SHG culture was 1 349.17±215.50 at 24 h,1 220.24±124.53 at 48 h and 1 982.14±219.03 at 72 h,all significantly lower than that in normal control at the corresponding time points(1 628.10±122.10,1 484.13±127.55 and 2 155.75±140.44,respectively,P<0.05),whereas it was obviously higher in FGC culture at 72 h(2 299.60±88.35),showing that LDH leakage in FGC was significantly higher than that in SHG at the corresponding time points(P<0.05).The LDH leakage was obviously decreased by Chinese medicine intervention with S-NSAB both in SHG at 72 h(1 797.62±146.40)and in FGC at 48 h(1 259.92±87.74)and 72 h(1 940.40±155.47),the difference between pre-and post-intervention was significant(P<0.05).Conclusion Fluctuated glucose conditions of culture medium could obviously damage the membranous stability of RGCs to enhance their permeability and lower the activity of cells;S-NSAB could improve these abnormalities in either SHG or FGC condition,which may be one of the important mechanisms of Chinese formula for nourishing Shen and activating blood in preventing and treating diabetic retinopathy.

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