清胰汤通过lncRNA-PVT1竞争性结合miR-146a靶向TRAF6参与重症胰腺炎发生发展的机制研究

Mechanism Study on the Role of Qingyi Tang in the Development of Severe Pancreatitis by Competitively Binding lncRNA-PVT1 to miR-146a Targeting TRAF6

目的探讨清胰汤通过lncRNA-PVT1竞争性结合miR-146a靶向肿瘤坏死因子受体相关因子6(TRAF6)参与重症胰腺炎发生发展的机制。方法购置SD大鼠50只,随机取10只设为空白对照组;剩余40只大鼠经肠壁穿刺逆行胰胆管注射5%牛磺胆酸钠注射制备重症胰腺炎动物模型,建模成功后随机取10只为模型对照组;剩余30只大鼠利用慢病毒转染干扰lncRNA-PVT1序列(LV-shPVT1)及空载(LV-control),分为清胰汤组、清胰汤+空载组和清胰汤+PVT1沉默组,每日用药1次,比较各组HE染色、炎症因子及TRAF6及细胞凋亡相关基因表达。结果HE染色结果表明,清胰汤组病理学改善显著,胰腺组织内炎症细胞浸润减轻明显;清胰汤+空载组和模型组可见微血栓形成,且随着时间延长加重,炎症细胞浸润明显,伴有上皮细胞浸润。清胰汤组TNF-α、IL-1β、TRAF6表达水平均低于清胰汤+空载组和清胰汤+PVT1沉默组(均P<0.05);清胰汤组miR-146a表达水平高于清胰汤+空载组和清胰汤+PVT1沉默组(P<0.05);清胰汤组细胞凋亡相关基因β-actin、ki-67、Bax、caspase、Caspase-9表达水平低于清胰汤+空载组和清胰汤+PVT1沉默组(均P<0.05)。结论在重症胰腺炎中,清胰汤能通显著下调lncRNA-PVT1表达,促进miR-146a表达,抑制TRAF6的表达,作用于炎症信号通路,降低炎症因子水平,抑制细胞凋亡,抑制重症胰腺炎的病理进展。

Objective To explore the mechanism of Qingyi Tang participating in the occurrence and development of severe pancreatitis by competitively binding lncRNA-PVT1 to miR-146a and targeting tumor necrosis factor receptor related factor 6(TRAF6).Methods A total of 50 SD rats were purchased and 10 were randomly selected as a blank control group;The remaining 40 rats were injected with 5%sodium taurocholate through intestinal wall puncture and retrograde pancreaticobiliary duct to establish a severe pancreatitis animal model.After successful modeling,10 rats were randomly selected as the model control group;The remaining 30 rats were divided into Qingyi Tang group,Qingyi Tang+empty load group,and Qingyi Tang+PVT1 silent group using lentivirus transfection to interfere with lncRNA-PVT1 sequence(LV shPVT1)and empty load(LV control).They were treated once a day and compared with HE staining,inflammatory factors,TRAF6,and apoptosis related gene expression in each group.Results The HE staining results showed that the pathological improvement was significant in the Qingyi Tang group,and the infiltration of inflammatory cells in the pancreatic tissue was significantly reduced;Micro thrombus formation can be observed in the Qingyi Tang+empty load group and model group,and it worsens with time,with significant infiltration of inflammatory cells accompanied by epithelial cell infiltration;the levels of TNF-α、IL-1βand TRAF6 in Qingyi Tang group were lower than that of Qingyi Tang+empty load group and Qingyi Tang+PVT1 silent group(P<0.05);The expression level of miR-146a in Qingyi Tang group was higher than that in Qingyi Tang+empty load group and Qingyi Tang+PVT1 silent group(P<0.05);The expression levels of cell apoptosis related genesβ-actin、ki-67,Bax,caspase,and Caspase-9 were lower than those of Qingyi Tang+empty group and Qingyi Tang+PVT1 silent group(P<0.05).Conclusions In severe pancreatitis,Qingyi Tang can significantly downregulate the expression of lncRNA-PVT1,enhance the expression of miR-146a,inhibit the expression of TRAF6,which can reduce inflammatory factor levels,inhibit cell apoptosis,and inhibit the pathological progress of severe pancreatitis.