越橘花青素对人胚胎巩膜成纤维细胞MMP2和collagen I表达的影响

 目的：探讨越橘花青素对人胚胎巩膜成纤维细胞（HFSF）基质金属蛋白酶2（MMP2）和I型胶原蛋白（collagen I）表达的影响，为越橘花青素防治近视提供实验依据。方法：取不同浓度（0、10^-6、10^-5、10^-4、10^-3、10^-2、10^-1和1 g/L）越橘花青素作用于HFSF，应用MTT法检测越橘花青素作用不同时间（6 h、8 h、12 h、24 h和48 h）对HFSF活力的影响；流式细胞术检测HFSF活力最高时（10^-1 g/L越橘花青素作用12 h）的细胞周期和凋亡情况；RT-qPCR检测HFSF中MMP2、COL1A1和COL1A2的mRNA表达；Western blot检测其MMP2和collagen I的蛋白表达。结果：MTT法结果显示HFSF在10^-1 g/L的越橘花青素作用12 h时活力最高；与空白对照组比较，流式细胞术显示10^-1 g/L越橘花青素组S和G₂期的细胞比例增加（P<0.05），凋亡率差异无统计学意义；RT-qPCR显示MMP2的mRNA表达量下降（P<0.05），COL1A1的mRNA表达量增加（P<0.05），COL1A2的mRNA表达量差异无统计学意义；Western blot显示MMP2蛋白表达下降（P<0.05），collagen I蛋白表达增加（P<0.05）。结论：越橘花青素对HFSF和具有抑制MMP2和增加collagen I表达的作用。

Effects of bilberry anthocyanins on MMP2 and collagen I in human fetal scleral fibroblast in vitro

AIM:To investigate the effects of bilberry anthocyanins on matrix metalloproteinase 2 (MMP2) and collagen type I (collagen I) expression in human fetal scleral fibroblasts (HFSF) in vitro and to provide experimental data for the prevention and treatment of myopia by bilberry anthocyanins. METHODS:HFSF were treated with bilberry anthocyanins at different concentrations (0, 10^-6, 10^-5, 10^-4, 10^-3, 10^-2, 10^-1 and 1 g/L). The effects of bilberry anthocyanins on the viability of HFSF for different time (6 h, 8 h, 12 h, 24 h and 48 h) were measured by MTT assay. The cell cycle distribution and apoptosis of HFSF with highest viability (10^-1 g/L bilberry anthocyanins for 12 h) were analyzed by flow cytometry. The mRNA expression of MMP2, COL1A1 and COL1A2 in the HFSF was detected by RT-qPCR. The protein expression of MMP2 and collagen I was determined by Western blot. RESULTS:The cell viability was the highest after treatment with bilberry anthocyanins at a concentration of 10^-1 g/L for 12 h. Compared with blank control group, 10^-1 g/L bilberry anthocyanin group showed increased cell numbers in S and G₂ phases (P<0.05), but no significant difference of the apoptotic rate was observed. The mRNA expression of MMP2 was decreased (P<0.05), and that of COL1A1 was increased (P<0.05). No significant difference of COL1A2 expression was seen. The protein expression of MMP2 was decreased (P<0.05), and collagen I protein was increased (P<0.05). CONCLUSION:Bilberry anthocyanins inhibit the expression of MMP2 and increase the expression of collagen I in the HFSF.