己糖激酶在原代培养腹膜间皮细胞的表达

Expression of Hexokinase in Peritoneal Mesothelial Cell of Primary Culture

目的 :探讨大鼠腹膜间皮细胞 (peritonealmesothelialcell,PMC)的原代培养方法及其己糖激酶 (hexoki nase ,HK)的表达。方法 :胰蛋白酶 EDTA消化法进行PMC的分离、培养 ,6 磷酸葡萄糖脱氢酶 (G6PDH)偶联比色法检测其HK总活性。结果 :PMC的原代及传代细胞 5～ 8d即可达到融合 ,其HK的基础活性为 (4.80± 0 .39)U/g蛋白。结论 :改良的胰蛋白酶消化法是进行PMC原代培养的一种可靠的实验方法。PMC有较强的HK活性 ,参与了腹膜透析时对葡萄糖吸收和利用方面的调节 。

﨩bjective: [WT5'BZ]Our aims were to study the prim ar y culture of rat peritoneal mesothelial cell (PMC) and to evaluate the expressio n of hexokinase (HK) activity. [WT5'HZ]Methods: [WT5'BZ]We obtained the PMC by using enzymatic disaggregation. Standard G6PDH coupled assay was used to measu re the total activity of HK. [WT5'HZ]Results: [WT5'BZ]It was approximately 5 to 8 days to reach confluency for primary and passaged cells. The basic activity of HK in PMC of rat was (4.80±0.39) U/g protein. [WT5'HZ]Conclusion: [WT5'BZ ]Trypsin EDTA treatment is a reproducible technique for the primary culture of peritoneal mesothelial cell. The expression of HK in PMC suggested that HK take part in the regulation of the glucose metabolism duing peritoneal dialysis and h ave a great effect on peritoneal dialysis ultrafilatration.