Xiu Caimei, Jiang Jing, Song Ruiying
Department of Ophthalmology, The Affiliated Yantai Yuhuangding Hospital of Qingdao University, Yantai, Shandong 264000, P.R. China.
Exp Ther Med. 2020 Feb;19(2):1051-1057. doi: 10.3892/etm.2019.8295. Epub 2019 Dec 5.
Expression of miR-34a in cataract rats and its related mechanism were investigated. A total of 30 SD rats were selected and divided into three groups: group A: 2-month-old lucent lens, group B: 18-month-old lucent lens, and group C: 18-month-old naturally occurring cataract lens. The lens was taken and measured by LOC III to determine the degree of lens opacity of the three groups of rats. qPCR was used to detect expression of miR-34a and mRNA of SIRT1 and P53. Western blotting was used to detect the protein expression of SIRT1 and P53. Cell apoptosis was detected by flow cytometry. The lens of rats in group C was more turbid than that of groups A and B (P<0.05). The expression levels of miR-34a and P53 mRNA in the rats lens of group C were significantly higher than those in groups A and B, and the expression of SIRT1 mRNA was significantly lower than that of groups A and group B (P<0.05). Expression of miR-34a in group A was significantly higher than that in group B, the mRNA expression of SIRT1 was significantly lower than that in the lucent lens of 18-month-old rats (P<0.05). The expression of SIRT1 protein in group C was significantly lower than that in groups A and group B, while the expression level of P53 protein in group C was significantly higher than that of groups A and B. The expression of SIRT1 protein in group B was significantly higher than that in group A (P<0.05). The apoptosis rate of group C was higher than that of groups A and group B (P<0.05). In conclusion, the upregulation of expression level of miR-34a is related to cataract occurrence in rats, which may be caused by regulation of SIRT1 protein.
研究了miR-34a在白内障大鼠中的表达及其相关机制。选取30只SD大鼠,分为三组:A组:2月龄透明晶状体;B组:18月龄透明晶状体;C组:18月龄自然发生白内障的晶状体。取晶状体用LOC III测量,以确定三组大鼠晶状体混浊程度。采用qPCR检测miR-34a表达及SIRT1和P53的mRNA。采用蛋白质免疫印迹法检测SIRT1和P53的蛋白表达。通过流式细胞术检测细胞凋亡。C组大鼠晶状体比A组和B组更混浊(P<0.05)。C组大鼠晶状体中miR-34a和P53 mRNA的表达水平显著高于A组和B组,SIRT1 mRNA的表达显著低于A组和B组(P<0.05)。A组miR-34a表达显著高于B组,SIRT1的mRNA表达显著低于18月龄透明晶状体大鼠(P<0.05)。C组SIRT1蛋白表达显著低于A组和B组,而C组P53蛋白表达水平显著高于A组和B组。B组SIRT1蛋白表达显著高于A组(P<0.05)。C组凋亡率高于A组和B组(P<0.05)。综上所述,miR-34a表达水平上调与大鼠白内障发生有关,可能是由SIRT1蛋白调控所致。