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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 x MAP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。
Objective To investigate the changes of proliferation and secretion of kupffer cells (KCs) in high glucose environment. Methods After primary KCs were cultured and expanded, the reference literatures were randomly divided into normal control group (11.1 mmol / L D-glucose), low glucose group (5.6 mmol / L D-glucose) D-glucose) and high glucose (25.0 mmol / L D-glucose). After culturing for 24, 48 and 72 h, the counting of blood cells was counted, the cell cycle was detected by flow cytometry, The cell proliferation was measured by colorimetric method and the cell supernatant was collected. Luminex x MAP was used to detect the expression of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) 6 levels. Results At 24, 48 and 72 h, the numbers of KCs and the OD value of KCs in high glucose group and low glucose group were significantly lower than those in control group and Chinese sugar group (P <0.01, P <0.05). The G0 / G1 phase in high glucose group and low glucose group at 24 h after culture was significantly higher than that in control group and medium sugar group, and the S phase and G2 / M phase were significantly lower than those in control group and Chinese sugar group (P <0.01). The levels of TNF-α, IL-1β and IL-6 in high glucose group were significantly lower than those in control group at 24, 48 and 72 h (P <0.05). Conclusions The activity of KCs in high glucose environment decreased obviously, and the proliferation and secretion function were inhibited.