中华眼底病杂志

中华眼底病杂志

糖尿病视网膜病变患者血浆miR-1470表达变化及可能机制探讨

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目的观察探讨糖尿病视网膜病变(DR)患者血浆中miR-1470的表达变化及可能机制。方法住院治疗的30例DR患者(DR组)以及在眼科门诊检查的30例糖尿病患者(DM组)、30名正常健康者(正常组)纳入研究。抽取三组受检者静脉血5 ml,提取血浆总RNA,纯化。通过基因芯片筛选出在DR患者血浆中具有差异表达的miRNA,并采用逆转录聚合酶链反应(RT-PCR)验证基因芯片检测结果。采用生物信息学预测miRNA调控的潜在靶基因,筛选出miR-1470及其靶基因表皮生长因子受体(EGFR)。将人视网膜内皮细胞(hREC)分为正常组(糖浓度5.5 mmol/L)、高糖组(糖浓度25.0 mmol/L)。将hREC转染miR-1470模拟物建立miR-1470高表达细胞模型,并将其分为空白对照组、高表达组、阴性对照组。采用RT-PCR检测细胞中miR-1470表达;采用蛋白免疫印迹法(Western blot)检测细胞中EGFR蛋白表达。两组间计量资料比较采用独立样本t检验,多组间计量资料比较采用方差分析, 组间计量资料两两比较采用多重比较。结果RT-PCR验证结果与基因芯片检测结果相符。DR组、DM组及正常组受检者血浆中miR-1470表达比较,差异有统计学意义(F=63.486,P=0.049)。与DM组和正常组比较,DR组患者血浆中miR-1470表达明显减弱,差异有统计学意义(q=111.2、73.9,P<0.05)。高糖组hREC中miR-1470表达明显低于正常组,差异有统计学意义(t=42.082,P=0.015)。高糖组hREC中EGFR蛋白表达明显高于正常组,差异有统计学意义(t=−39.939,P=0.016)。空白对照组、阴性对照组、高表达组hREC中miR-1470表达(F=637.069,P=0.000)、EGFR蛋白表达比较,差异有统计学意义(F=122.908,P=0.000)。与空白对照组、阴性对照组比较,高表达组hREC中miR-1470表达明显升高(q=329.7、328.8,P<0.05),EGFR蛋白表达明显降低(q=242.5、234.6,P<0.05),差异均有统计学意义。阴性对照组与空白对照组hREC中miR-1470表达、EGFR蛋白表达比较,差异无统计学意义(q=1.5、7.9,P>0.05)。结论DR患者血浆中miR-1470表达明显下调,EGFR表达增加可能与其有关。

ObjectiveTo investigate the expression and mechanism of miR-1470 in plasma of diabetic retinopathy (DR) patients.MethodsThirty patients with DR (DR group), 30 patients with diabetes (DM group) and 30 normal healthy subjects (normal group) were enrolled in the study. Three groups of subjects were taken 5 ml of venous blood, and total plasma RNA was extracted and purified. The differentially expressed miRNAs in the plasma of DR patients were screened by gene chip, and the results of gene chip detection were verified by reverse transcription polymerase chain reaction (RT-PCR). Bioinformatics was used to predict potential target genes for miRNA regulation, and miR-1470 and its target gene epidermal growth factor receptor (EGFR) were screened. Human retinal microvascular endothelial cells (hREC) were divided into normal group (sugar concentration 5.5 mmol/L) and high glucose group (sugar concentration 25.0 mmol/L). hREC was transfected into miR-1470 mimics to establish a miR-1470 high expression cell model, which was divided into blank control group, high expression group and negative control group. The expression of miR-1470 was detected by RT-PCR. The expression of EGFR protein was detected by Western blot. The measurement data of the two groups were compared using the independent sample t test. The comparison of the measurement data between the two groups was analyzed by ANOVA. The comparison between the measurement data of the groups was compared by multiple comparisons.ResultsThe results of RT-PCR were consistent with those of the gene chip. The expression of miR-1470 in the plasma of the DR group, the DM group and the normal group was statistically significant (F=63.486, P=0.049). Compared with the DM group and the normal group, the expression of miR-1470 in the DR group was significantly decreased, and the difference was statistically significant (q=111.2, 73.9; P<0.05). The expression of miR-1470 in hREC in the high glucose group was significantly lower than that in the normal group (t=42.082, P=0.015). The expression of EGFR protein in hREC of high glucose group was significantly higher than that of normal group (t=−39.939, P=0.016). The expression of miR-1470 (F=637.069, P=0.000) and EGFR (F=122.908, P=0.000) protein expression in hREC of blank control group, negative control group and high expression group were statistically significant . Compared with the blank control group and the negative control group, the expression of miR-1470 in hREC of high expression group was significantly increased (q=329.7, 328.8; P<0.05), and the expression of EGFR protein was significantly decreased (q=242.5, 234.6; P<0.05). There was no significant difference in the expression of miR-1470 and EGFR protein in hREC between the negative control group and the blank control group (q=1.5, 7.9; P>0.05).ConclusionThe expression of miR-1470 in the plasma of patients with DR is significantly down-regulated, and the increase of EGFR expression may be related to it.

关键词: 糖尿病视网膜病变/病因学; 微RNAs; 受体,表皮生长因子; 基因转移技术

Key words: Diabetic retinopathy/psychology; MicroRNAs; Receptor, epidermal growth factor; Gene transfer techniques

引用本文: 殷晓雯, 邵珺, 邹健, 谢田华, 姚勇. 糖尿病视网膜病变患者血浆miR-1470表达变化及可能机制探讨. 中华眼底病杂志, 2018, 34(4): 348-351. doi: 10.3760/cma.j.issn.1005-1015.2018.04.008 复制

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1. Ye EA, Steinle JJ. miR-146a suppresses STAT3/VEGF pathways and reduces apoptosis through IL-6 signaling in primary human retinal microvascular endothelial cells in high glucose conditions[J].Vision Res, 2017, 139:15-22.DOI:10.1016/j.visres.2017.03.009.
2. Ye EA, Steinle JJ. miR-146a attenuates inflammatory pathways mediated by TLR4/NF-kappaB and TNFalpha to protect primary human retinal microvascular endothelial cells grown in high glucose[J/OL]. Mediators Inflamm, 2016, 2016:3958453[2016-02-21] . https://dx.doi.org/10.1155/2016/3958453. DOI:10.1155/2016/3958453.
3. Lin X, Zhou X, Liu D,et al. MicroRNA-29 regulates high-glucose-induced apoptosis in human retinal pigment epithelial cells through PTEN[J].In Vitro Cell Dev Biol Anim, 2016, 52(4):419-426. DOI:10.1007/s11626-015-9990-z.
4. Gomaa AR, Elsayed ET, Moftah RF. MicroRNA-200b expression in the vitreous humor of patients with proliferative diabetic retinopathy[J]. Ophthalmic Res, 2017, 58(3):168-175. DOI:10.1159/000475671.
5. Barutta F, Bruno G, Matullo G.MicroRNA-126 and micro-/macrovascular complications of type 1 diabetes in the EURODIAB Prospective Complications Study[J].Acta Diabetol, 2017, 54(2):133-139. DOI:10.1007/s00592-016-0915-4.
6. 秦时月, 殷丽, 姚勇, 等. 糖尿病视网膜病变患者血清中微小RNA-195表达结果检测[J]. 中华眼底病杂志, 2015, 31(2):134-138. DOI: 10.3760/cma.j.issn.1005-1015.2015.02.006.Qin SY, Yin L, Yao Y, et al. The expression and role of miR-195 in diabetic retinopathy[J]. Chin J Ocul Fundus Dis, 2015,31(2):134-138. DOI:10.3760/cma.j.issn.1005-1015.2015.02.006.
7. Mei LL, Qiu YT, Wang WJ,et al.Overexpression of microRNA-1470 promotes proliferation and migration, and inhibits senescence of esophageal squamous carcinoma cells[J]. Oncol Lett, 2017, 14(6):7753-7758. DOI:10.3892/ol.2017.7190.
8. Murphrey MB, Bhimji SS. Biochemistry, epidermal growth factor receptor[J/OL]. StatPearls, 2018, 2018:E1[2018-01-19] .https://www.ncbi.nlm.nih.gov/books/NBK482459/.[published online ahead of print].
9. You H, Meng K, Wang ZY,et al.The ER-alpha36/EGFR signaling loop promotes growth of hepatocellular carcinoma cells[J]. Steroids, 2018, 134:78-87. DOI:1016/j.steroids.2018.02.007.
10. Zhang Y, Wei Y, Li X,et al. microRNA-874 suppresses tumor proliferation and metastasis in hepatocellular carcinoma by targeting the DOR/EGFR/ERK pathway[J]. Cell Death Dis, 2018, 9(2):130. DOI:10.1038/s41419-017-0131-3.
11. Chamberlain JJ, Johnson EL, Leal S,et al. Cardiovascular disease and risk management: review of the American Diabetes Association Standards of Medical Care in Diabetes 2018[J].Ann Intern Med, 2018, 168(9):640-650.DOI:10.7326/M18-0222.
12. 陈喆, 张士胜, 朱惠敏. 糖尿病视网膜病变的国际临床分类分析[J].国际眼科杂志, 2011, (8):1394-1401. DOI:10.3969/j.issn.1672-5123.2011.08.025.Chen Z, Zhang SS,Zhu HM, et al. Analysis of international clinical diabetic retinopathy disease severity scale[J].Int Eye Sci, 2011,11(8):1394-1401. DOI:10.3969/j.issn.1672-5123.2011.08.025.
13. 钱红, 胡柯, 刘理静, 等. miR-7靶向沉默EGFR抑制大鼠星形胶质细胞活化[J]. 中国药理学通报, 2018, 34(3):376-382. DOI: 10.3969/j.issn.1001-1978.2018.03.016.Qian H, Hu K, Liu LJ,et al. miR-7 inhibits activation of astrocytes derived from rats via silencing of EGFR[J]. Chinese Pharmacological Bulletin, 2018, 34(3): 376-382. DOI: 10.3969/j.issn.1001-1978.2018.03.016.