高糖环境对小鼠肝脏枯否细胞增殖及分泌功能的影响

目的：探讨高糖环境下枯否细胞( kupffer cells, KCs)增殖及分泌功能的变化。方法将小鼠原代KCs培养扩增后,参照文献随机分为正常对照组(11.1 mmol/L D-葡萄糖)、低糖组(5.6 mmol/L D-葡萄糖)、中糖组(12.5 mmol/L D-葡萄糖)和高糖组(25.0 mmol/L D-葡萄糖),培养24、48和72 h后,血球计数板进行细胞计数,流式细胞仪检测细胞周期,四甲基偶氮唑盐比色法测定细胞增殖,并收集细胞上清液,应用Luminex xMAP技术检测肿瘤坏死因子-α( tumornecrosis factor-α, TNF-α)、白介素-1β( interleukin-1β, IL-1β)和 IL-6水平。结果培养24、48和72 h时,高糖组和低糖组KCs扩增数量、OD值明显低于对照组和中糖组(P<0．01,P<0．05)。培养24 h时高糖组和低糖组G0/G1期明显高于对照组和中糖组,S期和G2/M期明显低于对照组和中糖组(P<0．01)。培养24、48和72 h时高糖组TNF-α、IL-1β和IL-6水平明显低于对照组(P<0．05)。结论高糖环境下KCs活性明显下降,增殖及分泌功能受到抑制。

Effects of High Glucose Environment on Proliferation and Secretion Function of Kupffer Cells in Liver of Mice

Objective To investigate changes of proliferation and secretion function of Kupffer cells ( KCs) in liver of mice under high glucose environment. Methods Primary KCs of mice were cultivated and proliferated, and then reference literature was divided into control group (11. 1 mmol/L D-glucose), low glucose group (LG, 5. 6 mmol/L D-glucose), medial glucose group (MG, 12. 5 mmol/L D-glucose) and high glucose group (HG, 25. 0 mmol/L D-glu-cose). After being cultured for 24, 48 and 72 h, KCs were counted by hemocytometer, and cell cycles were detected by flow cytometry, and cell proliferations were detected by methyl thiazolyl tetrazolium ( MTT) colorimetry respectively. Cell supernatant was collected, and levels of tumor necrosis factor-α ( TNF-α) , interleukin-1β ( IL-1β) and interleukin-6 ( IL-6) were detected by Luminex xMAP technique. Results After being cultured for 24, 48 and 72 h, numbers of KCs proliferation and optical density difference ( OD) values in HG and LG groups were significantly less than those in control and MG groups (P<0. 01, P<0. 05). After being cultured for 24 h, in HG and LG groups, KCs in G0/G1 phase were significantly more than those in control and MG groups, while KCs in S and G2/M phases were significantly less than those in control and MG groups (P<0. 01). Levels of TNF-α, IL-1βand IL-6 in HG group were significantly lower than those in control group after being cultured for 24, 48 and 72 h (P<0. 05). Conclusion KCs cytoactive is obviously de-crease, and proliferation and secretion function can be inhibited in mice under high glucose environment.