Gene expression analysis of effects of Met on BMDMs co-cultured with COM-TECs.

Microarray analysis was used to assess the expression levels of mRNAs in bone marrow-derived macrophages (BMDMs) pretreated with metformin (Met) or PBS and co-cultured with renal tubular epithelial cells (TECs) or COM-TECs (NC vs. COM, COM vs. COM + Met).BMDMs and TECs were isolated from wild-type (WT) C57BL/6J mice. To developed a BMDM-COM-stimulated TECs co-culture system, BMDMs were plated in the upper chamber of 6-well Transwell plates with a pore size of 0.4 µm (Corning, USA), while TECs were plated in the lower chamber. In the COM and COM + Met groups, TECs were treated with COM (100 µg/mL) for 24 h and BMDMs were treated with Met (5.0mM) for 24 h. Overall design: mRNA expression profile in BMDMs treated with Met or PBS and co-cultured with TECs or COM-TECs.

本研究采用基因微阵列分析（Microarray analysis），评估经二甲双胍（metformin, Met）或磷酸盐缓冲液（PBS）预处理，并分别与肾小管上皮细胞（renal tubular epithelial cells, TECs）或草酸钙晶体刺激的肾小管上皮细胞（COM-TECs）共培养的骨髓来源巨噬细胞（bone marrow-derived macrophages, BMDMs）的mRNA表达水平，实验分组设置为NC组 vs COM组、COM组 vs COM+Met组。BMDMs与TECs均分离自野生型（wild-type, WT）C57BL/6J小鼠。为构建BMDM-COM刺激的TECs共培养体系，将BMDMs接种于孔径为0.4 μm的6孔Transwell培养板的上室（美国Corning公司），TECs则接种于下室。在COM组与COM+Met组中，TECs经100 μg/mL的COM处理24小时，BMDMs经5.0 mM的Met处理24小时。整体实验设计：经二甲双胍或磷酸盐缓冲液预处理的BMDMs与肾小管上皮细胞或COM-TECs共培养后的mRNA表达谱。