高糖对血管内皮细胞层粘连蛋白α1的表达影响

Effect of high glucose on the expression of LAMA1 in vascular endothelial cells

目的分析高糖对视网膜血管内皮细胞中层粘连蛋白α1(LAMA1)的影响。方法体外培育视网膜血管内皮细胞(RF/6A),设置4种糖浓度培养液(5、15、25、35 mmol/L),其中5 mmol/L作为对照组,15、25、35 mmol/L作为高糖组。观察不同糖浓度培养液处理后的细胞生长情况,LAMA1的表达情况、检测细胞的迁移、增殖能力变化。结果随着观察时间的延长,高糖环境下细胞的生长速度增加,糖浓度越高,相对应生长速度越快。免疫荧光法检测发现,在处理24、48 h后,LAMA1在细胞质中表达,在高糖组(25、35 mmol/L)中表达量较对照组减少。处理24 h时,高糖组(25、35 mmol/L)中LAMA1蛋白表达量低于对照组(t分别=4.40、5.20,P均<0.05);处理48 h时,高糖组(35 mmol/L)LAMA1蛋白表达量和mRNA表达均低于对照组(t分别=6.80、4.23,P均<0.05)。在处理48、72 h后,高糖组(15、25、35 mmol/L)细胞的迁移能力较对照组明显增加(t分别=6.78、10.47、10.00、10.78、10.78、10.78,P均<0.05)。处理48、72、96 h后,高糖组(25、35 mmol/L)的细胞增殖能力明显高于对照组(t分别=6.68、4.60、3.89、4.09、4.29、4.37,P均<0.05)。结论高糖短期内刺激了细胞的生长、迁移、增殖,但抑制了LAMA1蛋白的表达,LAMA1可能参与了糖尿病视网膜病变的发生发展,并可能起到一定的保护作用。

Objective To analyze the effect of high glucose on the expression of LAMA1 in vascular endothelial cells.Methods Retinal vascular endothelial cells(RF/6A)were cultured in vitro,and four kinds of glucose concentrations were set up,among which 5 mmol/L as the control group,and 15 mmol/L,25 mmol/L,35 mmol/L as the high glucose group.The growth of cells,the expression of LAMA1,the migration and proliferation ability of cells were observed after being treated with different glucose concentrations.Results The growth rate of cells increased under high glucose environment,and the higher the glucose concentration,the faster the growth rate.Immunofluorescence detection showed that after 24h and 48h of treatment,LAMA1 was expressed in the cytoplasm,and the expression of LAMA1 in the high glucose group(25 mmol/L,35 mmol/L)was lower than that in the control group.After 24 h of treatment,the protein expression of LAMA1 in the high glucose group(25 mmol/L,35 mmol/L)was lower than that in the control group(t=4.40,5.20,P<0.05).After 48 h of treatment,the protein and mRNA expressions of LAMA1 in the high glucose group(35 mmol/L)were lower than those in the control group(t=6.80,4.23,P<0.05).After 48 h and 72 h of treatment,the migration ability of cells in the high glucose group(15 mmol/L,25 mmol/L,35 mmol/L)was significantly higher than that in the control group(t=6.78,10.47,10.00,10.78,10.78,10.78,P<0.05).After 48h,72h,96h of treatment,the cell proliferation ability of the high glucose group(25 mmol/L,35 mmol/L)was significantly higher than that of the control group(t=6.68,4.60,3.89,4.09,4.29,4.37,P<0.05).Conclusion High glucose stimulates the growth,migration and proliferation of cells in the short term,but inhibits the expression of LAMA1,which indirectly proves that LAMA1 may be involved in the occurrence and development of diabetic retinopathy,and may play a certain protective role.