山东医药

目的 观察环状RNA(circRNA)＿009396在心房颤动（简称房颤）患者外周血中的水平变化，并探讨circRNA＿009396过表达对人心房肌成纤维细胞（HCFs）纤维化的影响。方法 选择房颤患者44例作为房颤组，体检正常的健康志愿者44例作为无房颤组，采用实时荧光定量PCR法检测两组外周血circRNA＿009396水平，绘制外周血circRNA＿009396诊断房颤的受试者工作特征曲线并分析其诊断效能。取传至第3～5代的HCFs，加入20μmol/L的血管紧张素Ⅱ（AngⅡ）处理48 h以诱导纤维化，Western blotting法检测诱导后的细胞纤维化相关指标Ⅰ型胶原α1链（COL1A1）、Ⅲ型胶原α1链（COL3A1）、α平滑肌肌动蛋白（α-SMA）蛋白相对表达量均升高，证实成功诱导HCFs纤维化；将诱导纤维化的HCFs分为观察组和对照组，观察组加入circRNA＿009396过表达质粒DNA 2.5μg以及Lipo8000转染试剂4μL，对照组不予处理；采用实时荧光定量PCR法检测细胞circRNA＿009396及COL1A1、COL3A1、α-SMA mRNA相对表达量，Western blotting法检测细胞COL1A1、COL3A1、α-SMA蛋白相对表达量。结果 房颤组与无房颤组外周血circRNA＿009396水平分别为0.46±0.05、1.22±0.10，两组比较P<0.05。外周血circRNA＿009396诊断房颤的曲线下面积为0.902，截断值为0.542时的灵敏度为0.755、特异度为0.930。观察组细胞circRNA＿009396相对表达量高于对照组，COL1A1、COL3A1、α-SMA mRNA相对表达量均低于对照组（P均<0.05）。观察组细胞COL1A1、COL3A1、α-SMA蛋白相对表达量均低于对照组（P均<0.05）。结论 房颤患者外周血circRNA＿009396水平降低，过表达circRNA＿009396可抑制AngⅡ诱导的HCFs纤维化。

Objective To investigate the expression changes of circular RNA(circRNA)＿009396 in the peripheral blood of atrial fibrillation(AF) patients, and to explore the effects of circRNA＿009396 overexpression on fibrosis of Human cardiac fibroblasts(HCFs). Methods Forty-four patients with AF(AF group), and 44 healthy volunteers who received comprehensive physical examinations(non-AF group) were selected. Real-time fluorescence quantitative PCR was employed to measure the levels of circRNA＿009396 in peripheral blood samples from both groups. Subsequently, the receiver operating characteristic(ROC) curve for circRNA＿009396 in diagnosing AF was constructed, and its diagnostic efficacy was systematically evaluated. HCFs of the 3rd to 5th passages were treated with 20 μmol/L angiotensin Ⅱ （Ang Ⅱ） for 48 h to induce fibrosis. Western blotting confirmed that the relative expression levels of type Ⅰ collagen α1 chain(COL1A1), type Ⅲ collagen α1 chain(COL3A1), and α-smooth muscle actin(α-SMA), which were markers of cellular fibrosis, significantly increased, thereby validating the successful establishment of the fibrosis of HCFs. The the HCFs induced to fibrosis were subsequently divided into the observation group and control group. In the observation group, cells were transfected with 2. 5 μg circRNA＿009396 overexpression plasmid using 4 μL of Lipo8000 transfection reagent, whereas the control group remained untreated. Real-time fluorescence quantitative PCR was utilized to assess the relative mRNA expression levels of circRNA＿009396, COL1A1, COL3A1, and α-SMA in the cells, while Western blotting was performed to determine the relative protein expression levels of COL1A1, COL3A1, and α-SMA. Results The levels of circRNA＿009396 in the peripheral blood of the AF group and the non-AF group were 0. 46 ± 0. 05 and 1. 22 ± 0. 10, respectively, with statistically significant difference between the two groups(P<0. 05). The area under the curve(AUC) for diagnosing AF using peripheral blood circRNA＿009396 was 0. 902. When the cut-off value was set at 0. 542, the sensitivity reached 0. 755, and the specificity was 0. 930. Additionally, the relative expression level of circRNA＿009396 in the observation group was significantly higher than that in the control group(P<0. 05). Conversely, the relative expression levels of COL1A1, COL3A1, and α-SMA mRNA in the observation group were all significantly lower than those in the control group(all P<0. 05). Similarly, the protein expression levels of COL1A1, COL3A1, and α-SMA in the observation group were significantly lower than those in the control group(all P<0. 05). Conclusion The expression level of circRNA＿009396 in the peripheral blood of AF patients is significantly reduced and overexpression of circRNA＿009396may inhibit Ang Ⅱ-induced fibrosis of HCFs.