[1]韩莎莎,李跃峰,徐新萌.基于NF-κB信号通路探究miR-3197在糖尿病视网膜病变中的作用机制[J].眼科新进展,2024,44(3):188-192.[doi:10.13389/j.cnki.rao.2024.0037]
　HAN Shasha,LI Yuefeng,XU Xinmeng.Exploration of the mechanism of micro ribonucleic acid-3197 in diabetic retinopathy based on nuclear factor κB signaling pathway[J].Recent Advances in Ophthalmology,2024,44(3):188-192.[doi:10.13389/j.cnki.rao.2024.0037]
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基于NF-κB信号通路探究miR-3197在糖尿病视网膜病变中的作用机制/HTML
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
44卷
期数:
2024年3期
页码:
188-192
栏目:
实验研究
出版日期:
2024-03-05

文章信息/Info

Title:
Exploration of the mechanism of micro ribonucleic acid-3197 in diabetic retinopathy based on nuclear factor κB signaling pathway
作者:
韩莎莎; 李跃峰; 徐新萌
053000　河北省衡水市，衡水市人民医院眼科（韩莎莎，李跃峰）；210001　江苏省南京市，南京医科大学眼科医院（徐新萌）
Author(s):
HAN Shasha1; LI Yuefeng1; XU Xinmeng2
1.Department of Ophthalmology,Hengshui People’s Hospital,Hengshui 053000,Hebei Province,China
2.The Eye Hospital of Nanjing Medical University,Nanjing 210001,Jiangsu Province,China
关键词:
NF-κB信号通路; miR-3197; 糖尿病视网膜病变; 凋亡; 炎症因子
Keywords:
nuclear factor κB signaling pathway;  micro ribonucleic acid-3197;  diabetic retinopathy;  apoptosis;  inflammatory factor
分类号:
R774
DOI:
10.13389/j.cnki.rao.2024.0037
文献标志码:
A
摘要:
目的　基于核因子-κB（NF-κB）信号通路探究miR-3197在糖尿病视网膜病变（DR）中的机制。
方法　选取衡水市人民医院2021年1月至2021年12月收治的47例DR患者为DR组，并收集同期及同年龄段47例健康人作为对照组，收集其性别、年龄、空腹血糖（FBG）、空腹胰岛素（FINS）、甘油三酯（TG）、总胆固醇（TC）、miR-3197数据进行比较。分析DR患者miR-3197与实验室数据的相关性，并绘制miR-3197对DR诊断的受试者工作特征曲线（ROC曲线）。体外培养人视网膜微血管内皮细胞（hRMEC），分别采用5.5 mmol·L-1与30.0 mmol·L-1葡萄糖处理细胞，将其记为低糖（NG）组与高糖（HG）组；将anti-miR-NC与anti-miR-3197转染细胞后，加30.0 mmol·L-1葡萄糖处理，记为HG+anti-miR-NC组与HG+anti-miR-3197组。实时荧光定量PCR检测miR-3197相对表达量，流式细胞术检测hRMEC凋亡率，酶联免疫吸附法检测肿瘤坏死因子（TNF-α）与白细胞介素-6（IL-6）的表达，Western blot检测含半胱氨酸的天冬氨酸蛋白水解酶3（cleaved-caspase3）蛋白、Bax蛋白及NF-κB信号通路相关蛋白［磷酸化NF-κB p65（p-p65）、p65、磷酸化NF-κB抑制蛋白（p-IκBα）及NF-κB抑制蛋白（IκBα）］的表达。
结果　DR组患者FBG、FINS、TC及TG水平均高于对照组，差异均有统计学意义（均为P<0.001）。DR组患者外周血中miR-3197相对表达量（2.76±0.67）高于对照组（1.03±0.34），差异有统计学意义（P<0.05）。DR组患者miR-3197与FBG、FINS、TC及TG水平均呈正相关（r=0.672、0.587、0.511、0.423，均为P<0.05）。ROC曲线图显示，其曲线下面积为0.919，敏感度与特异度分别为85.11%与89.36%。与NG组相比，HG组细胞凋亡率、cleaved-caspase3、Bax、TNF-α、IL-6、p-IκBα及p-p65蛋白表达量均显著增加（均为P<0.05）；与HG+anti-miR-NC组相比，HG+anti-miR-3197组细胞凋亡率、cleaved-caspase3、Bax、TNF-α、IL-6、p-IκBα及p-p65蛋白表达量均显著降低（均为P<0.05）。
结论　miR-3197在DR患者外周血和高糖诱导的hRMEC中高表达，下调miR-3197可减轻高糖诱导的hRMEC凋亡及炎性损伤，其作用机制可能与抑制NF-κB信号通路有关。
Abstract:
Objective　To explore the mechanism of micro ribonucleic acid (miR)-3197 in diabetic retinopathy (DR) on the basis of the nuclear factor κB (NF-κB) signaling pathway.
Methods　A total of 47 DR patients admitted to Hengshui People’s Hospital from January 2021 to December 2021 were selected as the DR group, and 47 healthy individuals in the same period were collected as the control group. Their information in gender, age, fasting blood glucose (FBG), fasting insulin (FINS), triglycerides (TG), total cholesterol (TC) and miR-3197 were compared. The correlation between miR-3197 in DR patients and laboratory data was analyzed, and the receiver operating characteristic (ROC) curve of miR-3197 for DR diagnosis was drawn. The human retinal microvascular endothelial cells (hRMECs) were cultured in vitro and treated with 5.5 mmol·L-1 glucose ［low glucose (NG) group］ and 30 mmol·L-1 glucose ［high glucose (HG) group］, respectively. After transfecting with anti-miR-NC and anti-miR-3197, the cells were treated with 30 mmol·L-1 glucose (HG+anti-miR-NC group and HG+anti-miR-3197 group). Real-time fluorescence quantitative PCR was used to detect the relative expression level of miR-3197, flow cytometry was used to detect the apoptosis rate of hRMECs, enzyme-linked immunosorbent assay was used for detecting tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6), and Western blot was adopted to detect the expressions of aspartic protease 3 containing cysteine (cleaved caspase-3) protein, Bax protein and NF-κB signaling pathway-related proteins ［phospho-NF-κB p65 (p-p65), p65, phospho-NF-κB inhibited protein (p-IκBα), and NF-κB inhibited protein (IκBα)］.
Results　The levels of FBG, FINS, TC and TG in the DR group were higher than those in the control group, and the differences were statistically significant (all P<0.001). The relative expression level of miR-3197 in the peripheral blood of patients in the DR group (2.76±0.67) was higher than that of the control group (1.03±0.34), and the difference was statistically significant (P<0.05). The miR-3197 level of patients in the DR group was positively correlated with FBG, FINS, TC and TG levels (r=0.672, 0.587, 0.511 and 0.423; all P<0.05). The ROC curve graph showed that the area under the curve was 0.919, with sensitivity and specificity of 85.11% and 89.36%, respectively. Compared with the NG group, the HG group showed a significant increase in cell apoptosis rate and the protein expressions of cleaved caspase-3, Bax, TNF-α, IL-6, p-IκBα and p-p65 (all P<0.05); compared with the HG+anti-miR-NC group, the HG+anti-miR-3197 group showed a significant decrease in cell apoptosis rate and the protein expressions of cleaved caspase-3, Bax, TNF-α, IL-6, p-IκBα and p-p65 (all P<0.05).
Conclusion　The miR-3197 is highly expressed in the peripheral blood of DR patients and high glucose-induced hRMECs. Down-regulation of miR-3197 can alleviate high glucose-induced hRMEC apoptosis and inflammatory injury, and its mechanism of action may be related to the inhibition of the NF-κB signaling pathway.

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备注/Memo

备注/Memo:
河北省卫生健康委员会项目（编号：20191762）
更新日期/Last Update: 2024-03-05