Exploration of the mechanism of micro ribonucleic acid-3197 in diabetic reti-nopathy based on nuclear factor κB signaling pathway / 眼科新进展

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 Heng-shui 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),fast-ing 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 im-munosorbent assay was used for detecting tumor necrosis factor-a(TNF-a)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-KB p65(p-p65),p65,phospho-NF-KB 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 pro-tein expressions of cleaved caspase-3,Bax,TNF-a,IL-6,p-IκBa 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 expres-sions of cleaved caspase-3,Bax,TNF-a,IL-6,p-IκBa and p-p65(allP＜0.05).Conclusion The miR-3197 is highly ex-pressed in the peripheral blood of DR patients and high glucose-induced hRMECs.Down-regulation of miR-3197 can allevi-ate high glucose-induced hRMEC apoptosis and inflammatory injury,and its mechanism of action may be related to the inhi-bition of the NF-κB signaling pathway.