葡萄糖浓度波动对兔视网膜Müller细胞生长活性和iNOS表达的影响

The effects of glucose fluctuation on the viability and the expression of inducible nitric oxide synthase of rabbit retinal Müller cells in vitro

目的兔视网膜Müller细胞在葡萄糖浓度波动培养下,观察诱导型一氧化氮合酶(iNOS)、细胞生长活性的表达及变化。方法酶消化法培养兔视网膜Müller细胞,分为4组,正常对照组(N),持续高糖组(HG),葡萄糖浓度波动组(GF),渗透压对照组(OP);四甲基偶氮唑盐比色法(MTT)测定各孔光吸收(OD)值。采用免疫荧光染色检测各组细胞iNOS平均荧光强度的改变。结果与N组比较,HG组和GF组OD值下降,iNOS表达升高,均有显著差异(p<0.01);与HG组比,GF组OD值下降(p<0.01),iNOS表达升高,有统计学差异(p<0.05);OP组与N组比较无明显差异(p>0.05)。结论与持续高糖相比Müller细胞在葡萄糖浓度波动时生长活性下降,iNOS表达升高。提示葡萄糖浓度波动在糖尿病视网膜病变发生发展中可能发挥一定的作用。

Objective To investigate the change of rabbit retinal Mtiller cells＇ viability and inducible nitric oxide synthase （iNOS） ＇ s expression in fluctuated glucose in vitro. Methods Milller cells were isolated by enzymatic digestion and cultured in vitro. The cells were divided into four groups： normal control （N）, continuous high glucose （HG）, glucose fluctuation （GF）, and osmolality control （OP） groups. MTT assay was used to detect the OD value of the MUller cells in the 96 - well plate ; iNOS expression in Mtiller cells was observed by immunofluorescence staining. Results Compared with the group of normal glucose, OD value was obviously decreased and iNOS expression was significantly enhanced in the contin-uous high glucose group and glucose fluctuation group（p 〈 0.01 ） ; glucose fluctuation group＇s OD value is lower than continuous high glucose group（p 〈 0.01 ） , but iNOS expression was increased more than continuous high glucose group（p 〈0.05） ; The control of osmotic pressure of culture solution group had no significant differences compared with the normal glucose group （p 〉 0.05）. Conclusions Glucose fluctuation can inhibit the proliferation of Mtiller cells and promote iNOS expression than continuous high level glucose. It indicates that glucose fluctuation may play certain role in the development of diabetic retinopathy.