目的 探讨氧化苦参碱（OM）是否通过胰腺星状细胞株（LTC-14）中miR-211-5p调节TLR4/NF-κB通路调控炎性反应。方法 用生物预测、文献分析筛选出目的miR；脂多糖（LPS）刺激LTC-14细胞制备炎症模型，通过实时荧光定量PCR（qRT-PCR）法检测miR的变化以及OM对其的调节作用；通过转染mimics和inhibitor来过表达和低表达目的miR后，以Western Blotting法、实时荧光定量PCR（qRT-PCR）法检测LTC-14细胞中TLR4/NF-κB信号通路相关分子（TLR4、MyD88、NF-κB）的表达情况，ELISA法检测细胞上清中炎性因子TNF-α的表达情况。结果 经生物信息学预测筛选出的目的miR为miR-211-5p；与对照组比较，LPS刺激LTC-14细胞后miR-211-5p表达明显下降（P<0.001）；与模型组比较，经OM干预后miR-211-5p表达显著上升（P<0.001）。过表达miR-211-5p后，TLR4、MyD88、NF-κB的mRNA和蛋白水平，以及炎性因子TNF-α表达显著下降（P<0.05、0.01、0.001）；低表达miR-211-5p后，TLR4、MyD88、NF-κB的mRNA和蛋白水平，以及炎性因子TNF-α表达上升（P<0.05、0.01、0.001）。结论 PSCs中的miR-211-5p通过调节TLR4/NF-κB信号通路调节炎症反应，OM可以调节炎症模型中miR-211-5p的表达，OM调节炎症反应的作用机制可能是通过miR-211-5p调节TLR4/NF-κB信号通路实现的。

Objective To investigate the anti-inflammatory mechanism of oxymatrine (OM) on targeting TLR4/NF-κB signaling pathwayby regulating miRNA-211-5p in LTC-14 cells. Methods The target miRNAs were screened by biological prediction and literature analysis. Lipopolysaccharide (LPS) stimulates LTC-14 cells to prepare inflammatory models, and miRNAs expression variation and the regulation of OM were detected by real-time fluorescence quantitative PCR (RT-qPCR). Up-regulation and down-expression of miRNAs were manufactured by mimics and inhibitor transfection. The expression of related molecules (TLR4, MyD88, and NF-κB) in TLR4/NF-κB signaling pathway of LTC-14 cells was detected by Western blotting and RT-qPCR. The expression of inflammatory factor TNF-α was detected by ELISA assay in cell supernatant. Results miRNA-211-5p was chosen as target for further study after bioinformatics screening. The expression of miR-211-5p in LTC-14 cells stimulated by LPS decreased significantly (P<0.001) compared with the control group. The decreased expression of miRNA-211-5p induced by LPS was promoted by OM administration (P<0.001) compared with the model group. After up-regulating of miRNA-211-5p, the expression of TLR4/NF-κB signaling pathway related molecules (TLR4, MyD88, and NF-κB) and TNF-αdecreased significantly (P<0.05, 0.01, and 0.001). After down-regulation of miRNA-211-5p, the expression of TLR4, MyD88 and NF-κB TNF-α were increased (P<0.05, 0.01, and 0.001). Conclusion OM can modulate TLR4/NF-κB inflammatory response signaling pathway by regulating the expression of miRNA-211-5p in the PSCs.

国家自然科学基金资助项目（81173393）；天津市应用基础与前沿技术计划项目（17JCYBJC26700）；新乡医学院研究生创新基金（YJSCX201651Y）