摘要
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) and 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48h)对HFSF活力的影响;流式细胞术检测HFSF活力最高时(10-1g/L越橘花青素作用12 h)的细胞周期和凋亡情况;RT-q PCR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10-1g/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10-1g/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-q PCR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。
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,COL1 A1 and COL1 A2 in the HFSF was detected by RT-q PCR. 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 COL1 A1 was increased（ P〈0. 05）. No significant difference of COL1 A2 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.
作者
刘丽敏
钟华红
邓宏伟
肖红萍
曾杞汶
田原
刘小勇
孟晶
LIU Li-min;ZHONG Hua-hong;DENG Hong-wei;XIAO Hong-ping;ZENG Qi-wen;TIAN Yuan;LIU Xiao-yong;MENG Jing(Department of Ophthalmology,The First Affiliated Hospital of Jinan University,Guangzhou 510632,China;ShenzhenEye Hospital Affiliated to Jinan University,Shenzhen 518040,China.)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2018年第7期1291-1296,共6页
Chinese Journal of Pathophysiology
基金
广东省中医药基金资助项目(No.20162043
No.20162044)
深圳市科技创新委基金资助项目(No.JCYJ20160428144325551
No.JCYJ20150402152130697
No.JCYJ20170306113900538)
