高糖环境下视网膜微血管周细胞β1,4半乳糖基转移酶-1的表达

Expression of β1,4 Galactosyltransferase-1 in Bovine Retinal Pericytes Cultured with High Glucose

目的了解高糖培养环境下牛视网膜微血管周细胞β1,4半乳糖基转移酶-1(galactosyltrans-ferase-1,GalT-1)的表达情况以及细胞凋亡情况。方法原代培养牛视网膜微血管周细胞,在含不同浓度葡萄糖的培养基(5 mmol/L,10 mmol/L,20 mmol/L,30 mmol/L)培养72 h,以5 mmol/L 浓度葡萄糖培养组作为正常对照组。采用 RT-PCR,蛋白凝集素染色法观察不同浓度葡萄糖培养下 GalT-1表达的改变情况,同时采用流式细胞观察高糖诱导的细胞凋亡。结果高糖培养环境下,视网膜微血管周细胞内 GalT-1 mRNA 表达上调,凝集素染色提示视网膜微血管周细胞蛋白由 Galβ 1→4GlcNAc 糖基化基团的修饰表达上调;视网膜微血管周细胞凋亡加剧,各葡萄糖浓度组的凋亡细胞平均死亡率为:7.35±1.82%,16.34±2.25%,29.85±6.66%,39.48±6.48%。结论高糖环境下,牛视网膜微血管周细胞内β1,4半乳糖基转移酶-1的表达上调,其可能参与了高糖环境下的视网膜微血管周细胞凋亡,参与机制有待进一步研究。(中国眼耳鼻喉科杂志,2007,7:147-149)

Purpose To investigate the apoptosis and expression of β1,4 galactosyhransferase-1 （ GalT-1 ） in bovine retinal pericytes （BRPs） which were cultured with high glucose. Methods BRPs were cultured in medium containing different concentration of glucose （5 mmol/L, 10 mmol/L, 20 mmol/L, 30 mmol/L） for 72 hours. Gene expression of GalT-1 in BRPs was observed with reverse transcription polymerase chain reaction. Lectin blot assay with RCA-I was performed to examine the level of the groups of Galβ1→4GlcNAc on cell total proteins, and the level of apoptosis of BRPs was examined with flow cytometry analysis. Results High level of glucose could increase the expression of GalT-1 at mRNA level and increase the groups of Galβ1→4GlcNAc in BRPs. Simultaneously, the rate of apoptosis of BRPs increased and the average rates of apoptosis in different concentration of glucose used are 7.35±1.82%, 16.34±2.25% ,29.85±6.66%. 39.48± 6. 48%, respectively. Conclusion High level of glucose up-regulates the expression of GalT-1 in BRPs, and the up-regulation of GalT-1 might be involved in apoptosis in the pericytes.