miR-217和SIRT1在老年白内障晶状体上皮细胞中的表达水平及意义

The expression relationship and significance of miR-217 and SIRT1 in lens epithelial cells of senile cataract

目的探讨微小RNA-217(miR-217)与沉默信息调节因子相关酶1(SIRT1)在老年白内障晶状体上皮细胞(LECs)中的表达关系及意义。方法选取2017年12月至2018年3月沧州市中心医院医疗集团收治的65例老年性白内障患者作为研究组,并根据不同类型将其分为皮质性组(30例)、核性组(22例)、后囊下组(13例),另选取同期于本院进行角膜移植术的其他眼部疾病(无白内障)患者38例为对照组。采用实时荧光定量PCR(RT-PCR)检测LECs中miR-217与SIRT1 mRNA表达水平;蛋白免疫印迹法(WB)检测SIRT1蛋白表达情况;采用酶联免疫吸附法(ELISA)检测血清超氧化物歧化酶(SOD)、丙二醛(MDA)、过氧化脂质(LPO)水平;采用ABTS比色法检测总抗氧化能力(TAC)。采用Pearson法进行相关性分析。结果与对照组相比,研究组miR-217表达水平较高,而SIRT1 mRNA及蛋白表达水平较低,差异有统计学意义(P<0.05)。研究组血清SOD、TAC水平均显著低于对照组,而MDA、LPO水平均高于对照组,差异有统计学意义(P<0.05)。与皮质性组相比,核性组与后囊下组miR-217表达水平均显著升高,而SIRT1 mRNA及蛋白表达水平均显著降低,差异均有统计学意义(P<0.05)。Pearson结果显示,miR-217与SIRT1呈负相关,与MDA、LPO呈正相关,而与SOD、TAC呈负相关(P<0.05);SIRT1与SOD、TAC呈正相关(P<0.05),而与MDA、LPO呈负相关(P<0.05)。结论miR-217在老年白内障患者LECs中异常高表达,而SIRT1呈低表达,二者呈负相关且均可能通过参与老年白内障氧化应激反应过程进而促进疾病发生及发展。

Objective To analyze the expression relationship and significance of micro RNA-217(miR-217)and silent information regulator related enzyme 1(SIRT1)in lens epithelial cells(LECs)of senile cataract.Methods From December 2017 to March 2018,65 senile patients with cataract were selected as the study group;according to different types,they were divided into cortical group(30 cases),nuclear group(22 cases),and posterior subcapsular group(13 cases).Another 38 patients with other eye diseases(no cataract)who underwent corneal transplantation in our hospital during the same period were selected as the control group.Real-time fluorescence quantitative PCR(RT-PCR)was used to detect the expression levels of miR-217 and SIRT1 mRNA in LECs;Western blot(WB)was used to detect the expression of SIRT1 protein;the levels of serum superoxide dismutase(SOD),malondialdehyde(MDA),and lipid peroxide(LPO)were detected by enzyme-linked immunosorbent assay(ELISA);the total antioxidant capacity(TAC)was detected by ABTS colorimetry.Pearson method was used for correlation analysis.Results Compared with the control group,the expression of miR-217 in the study group was significantly higher than that in the control group(P<0.05),while the expressions of SIRT1 mRNA and protein were significantly lower than those in the control group(P<0.05);the levels of serum SOD and TAC in the study group were significantly lower than those in the control group(P<0.05),while the levels of MDA and LPO were significantly higher(P<0.05);compared with cortical group,the expression level of miR-217 in nuclear group and posterior subcapsular group increased significantly(P<0.05),while the expressions of SIRT1 mRNA and protein were decreased significantly(P<0.05);the Pearson method showed there was a significant negative correlation between miR-217 and SIRT1(P<0.05);miR-217 was positively correlated with MDA and LPO(P<0.05),but negatively correlated with SOD and TAC(P<0.05);SIRT1 was positively correlated with SOD and TAC(P<0.05),but negatively correlated with MDA and LPO(P<0.05).Conclusion MiR-217 was highly expressed in LECs of senile cataract patients,while SIRT1 was low expressed;they were negatively correlated and might contribute to the occurrence and development of cataract by participating in the oxidative stress process of senile cataract.