目的 探討氧化苦參堿（OM）是否通過胰腺星狀細(xì)胞株（LTC-14）中miR-211-5p調(diào)節(jié)TLR4/NF-κB通路調(diào)控炎性反應(yīng)。方法 用生物預(yù)測、文獻(xiàn)分析篩選出目的miR；脂多糖（LPS）刺激LTC-14細(xì)胞制備炎癥模型，通過實(shí)時熒光定量PCR（qRT-PCR）法檢測miR的變化以及OM對其的調(diào)節(jié)作用；通過轉(zhuǎn)染mimics和inhibitor來過表達(dá)和低表達(dá)目的miR后，以Western Blotting法、實(shí)時熒光定量PCR（qRT-PCR）法檢測LTC-14細(xì)胞中TLR4/NF-κB信號通路相關(guān)分子（TLR4、MyD88、NF-κB）的表達(dá)情況，ELISA法檢測細(xì)胞上清中炎性因子TNF-α的表達(dá)情況。結(jié)果 經(jīng)生物信息學(xué)預(yù)測篩選出的目的miR為miR-211-5p；與對照組比較，LPS刺激LTC-14細(xì)胞后miR-211-5p表達(dá)明顯下降（P<0.001）；與模型組比較，經(jīng)OM干預(yù)后miR-211-5p表達(dá)顯著上升（P<0.001）。過表達(dá)miR-211-5p后，TLR4、MyD88、NF-κB的mRNA和蛋白水平，以及炎性因子TNF-α表達(dá)顯著下降（P<0.05、0.01、0.001）；低表達(dá)miR-211-5p后，TLR4、MyD88、NF-κB的mRNA和蛋白水平，以及炎性因子TNF-α表達(dá)上升（P<0.05、0.01、0.001）。結(jié)論 PSCs中的miR-211-5p通過調(diào)節(jié)TLR4/NF-κB信號通路調(diào)節(jié)炎癥反應(yīng)，OM可以調(diào)節(jié)炎癥模型中miR-211-5p的表達(dá)，OM調(diào)節(jié)炎癥反應(yīng)的作用機(jī)制可能是通過miR-211-5p調(diào)節(jié)TLR4/NF-κB信號通路實(shí)現(xiàn)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.

國家自然科學(xué)基金資助項(xiàng)目（81173393）；天津市應(yīng)用基礎(chǔ)與前沿技術(shù)計(jì)劃項(xiàng)目（17JCYBJC26700）；新鄉(xiāng)醫(yī)學(xué)院研究生創(chuàng)新基金（YJSCX201651Y）