Shimoda Yoko, Tagaya Yuko, Saito Tsugumichi, Yamada Eijiro, Osaki Aya, Nakajima Yasuyo, Ozawa Atsushi, Satoh Tetsurou, Okada Junichi, Okada Shuichi, Yamada Masanobu
Department of Medicine and Molecular Science, Gunma University Graduate School of Medicine, Showa-machi, Maebashi, Gunma, Japan.
Cell J. 2017 Spring;19(Suppl 1):106-112. doi: 10.22074/cellj.2017.4446. Epub 2017 May 17.
We studied effect of high glucose levels on coronary artery endothelial cell proliferation and human colon cancer cell proliferation. To examine the long-term effect of glucose exposure on cell growth, cells were cultured for 14 days in the absence or presence of 183 mg/dL D-glucose addition in the culture medium. Short effect of elevated glucose levels was examined by addition of 183 mg/dL D-glucose addition in the culture medium for just one hour per day followed by changing the culture to standard medium (5.5 mM D-glucose) during the next 23-hours period. Cell proliferation was estimated by 2,3-Bis (2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carbox-anilide (XTT) assay and phosphor-Erk western blot analysis. We found that coronary artery endothelial cell proliferation was significantly increased in the culture medium with the acute one-hour addition of 183 mg/dL D-glucose compared to the absence or chronic presence of 183 mg/dL D-glucose addition in the culture medium. In contrast, colon cancer cell proliferation was significantly increased in the continuous presence of 183 mg/dL D-glucose addition in the culture medium compared to the acute one-hour addition of glucose. The extent of Erk2 phosphorylation paralleled with the relative changes in cellular proliferation in both cell types. Taken together, these results suggested that continuous or transient high level of glucose exposure differentially effects coronary artery endothelial and human colon cancer cell proliferation.
我们研究了高糖水平对冠状动脉内皮细胞增殖和人结肠癌细胞增殖的影响。为了检测葡萄糖暴露对细胞生长的长期影响,将细胞在无或添加183mg/dL D-葡萄糖的培养基中培养14天。通过在培养基中每天仅添加183mg/dL D-葡萄糖1小时,然后在接下来的23小时内将培养基更换为标准培养基(5.5mM D-葡萄糖),来检测葡萄糖水平升高的短期影响。通过2,3-双(2-甲氧基-4-硝基-5-磺基苯基)-2H-四唑-5-甲酰苯胺(XTT)分析和磷酸化-Erk免疫印迹分析来评估细胞增殖。我们发现,与在培养基中不存在或长期存在183mg/dL D-葡萄糖相比,在培养基中急性添加183mg/dL D-葡萄糖1小时后,冠状动脉内皮细胞增殖显著增加。相反,与急性添加葡萄糖1小时相比,在培养基中持续存在183mg/dL D-葡萄糖时,结肠癌细胞增殖显著增加。两种细胞类型中Erk2磷酸化程度与细胞增殖的相对变化平行。综上所述,这些结果表明,持续或短暂的高糖暴露对冠状动脉内皮细胞和人结肠癌细胞增殖的影响不同。