Department of Emergency, The Second People's Hospital of Guiyang, Guiyang, 550023, China.
Department of Intensive Care Unit, The Second People's Hospital of Guiyang, Guiyang, 550023, China.
Pflugers Arch. 2019 Dec;471(11-12):1519-1527. doi: 10.1007/s00424-019-02329-5. Epub 2019 Nov 15.
Acute pancreatitis (AP) is a common clinical critical disease with high mortality and the exact pathogenesis is not fully elucidated. The present study aimed to uncover the function of miR-135a in the proliferation, apoptosis, and inflammatory characteristics of diseased pancreatic cells and the potential molecular mechanisms. The expression patterns of miR-135a and family with sequence similarity 129 member A (FAM129A) in patients with AP were analyzed on the basis of the GEO database. The transfection efficiency and expression level of miR-135a in AR42J cells were determined by qRT-PCR. The biological characteristics of AR42J cells treated with cerulein were detected by cell counting kit-8 (CCK-8), flow cytometry, and western blot assays. The potential interaction between miR-135a and FAM129A was confirmed by bioinformatics prediction softwares and luciferase reporter assay. MiR-135a inhibitor and pcDNA3.1-FAM129A were co-transfected to determine the regulation of miR-135a/FAM129A on inflammatory AR42J cell injury. We observed that miR-135a was highly expressed in AP samples. Depletion of miR-135a could alleviate the condition so that the AR42J cells proliferation increased, apoptosis decreased, and the expression of inflammatory cytokines enhanced. In addition, mRNA and protein expression of FAM129A were negatively regulated by miR-135a, and over-expression of FAM129A could strengthen the relief effect of miR-135a inhibitor in AP induced by cerulein. In summary, our data demonstrates that silencing miR-135a reduces AR42J cells injury and inflammatory response in AP induced by cerulein through targeting FAM129A.
急性胰腺炎（AP）是一种常见的临床危重症疾病，死亡率高，其确切发病机制尚未完全阐明。本研究旨在揭示 miR-135a 在病变胰腺细胞增殖、凋亡和炎症特征中的功能及其潜在的分子机制。基于 GEO 数据库分析了 AP 患者 miR-135a 和家族与序列相似性 129 成员 A（FAM129A）的表达模式。通过 qRT-PCR 测定 AR42J 细胞中转染 miR-135a 的效率和表达水平。通过细胞计数试剂盒-8（CCK-8）、流式细胞术和 Western blot 检测细胞处理 cerulein 后的生物学特征。通过生物信息学预测软件和荧光素酶报告基因检测证实 miR-135a 与 FAM129A 之间的潜在相互作用。共转染 miR-135a 抑制剂和 pcDNA3.1-FAM129A 以确定 miR-135a/FAM129A 对炎症性 AR42J 细胞损伤的调节作用。我们观察到 miR-135a 在 AP 样本中高表达。miR-135a 的耗竭可以缓解这种情况，使 AR42J 细胞增殖增加，凋亡减少，炎症细胞因子的表达增强。此外，FAM129A 的 mRNA 和蛋白表达受 miR-135a 的负调控，而过表达 FAM129A 可以增强 cerulein 诱导的 AP 中 miR-135a 抑制剂的缓解作用。综上所述，我们的数据表明，沉默 miR-135a 通过靶向 FAM129A 减轻 cerulein 诱导的 AP 中 AR42J 细胞损伤和炎症反应。
