TMEM16A Ca2+-activated Cl- channel inhibition ameliorates acute pancreatitis via the IP3R/Ca2+/NFκB/IL-6 signaling pathway.

TMEM16A Ca2 + 激活的 Cl - 通道抑制通过 IP3R / Ca2 + / NF κ B / IL - 6 信号通路改善急性胰腺炎。

• 影响指数: 12.822
• 发表时间: May 2020
• 来源期刊: J Adv Res
• DOI: 10.1016/j.jare.2020.01.006
• PMID: 32071789
• 文章类型: Journal Article
• 作者列表: Wang Q,Bai L,Luo S,Wang T,Yang F,Xia J,Wang H,Ma K,Liu M,Wu S,Wang H,Guo S,Sun X,Xiao Q

关键词: AP, acute pancreatitis; Acute pancreatitis; BAPTA-AM, 1,2-bis(2-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid-acetyloxymethyl ester; CCK, cholesystokinin; CFBE, cystic fibrosis bronchial epithelial; CaCCinh-A01, Ca2+-activated Cl− channel inhibitor-A01; EDTA, ethylenediaminetetraacetic acid; EGF, epidermal growth factor; EGFP, green fluorescent protein; EGFR, epidermal growth factor receptor; EGTA, ethylene glycol-bis(2-aminoethyl ether)-N,N,N',N'-tetraacetic acid; ELISA, enzyme-linked immunosorbent assay; ER, endoplasmic reticulum; FBS, fetal bovine serum; HEPES, N-2-hydroxyethil-piperazine-N'-2-ethanesulfonic acid; IL-6, interleukin 6; IL-6R, interleukin 6 receptor; IP3R, inositol 1,4,5-trisphosphate receptor; Inositol 1,4,5-trisphosphate receptor; Interleukin-6; NFκB; NFκB, nuclear factor-κB; NMDG, N-methyl-D-glucamine; NP-40, Nonidet P-40; PACs, pancreatic acinar cells; RIPA, radio immunoprecipitation assay; SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis; STAT3, signal transducers and activators of transcription 3; T16Ainh-A01, TMEM16A inhibitor-A01; TMEM16A; Tris, tris(hydroxymethyl)aminomethane; WT, wild type; shRNAs, short hairpin RNAs

Abstract

TMEM16A Ca2+-activated Cl- channels are expressed in pancreatic acinar cells and participate in inflammation-associated diseases. Whether TMEM16A contributes to the pathogenesis of acute pancreatitis (AP) remains unknown. Here, we found that increased TMEM16A expression in the pancreatic tissue was correlated with the interleukin-6 (IL-6) level in the pancreatic tissue and in the serum of a cerulein-induced AP mouse model. IL-6 treatment promoted TMEM16A expression in AR42J pancreatic acinar cells via the IL-6 receptor (IL-6R)/signal transducers and activators of transcription 3 (STAT3) signaling pathway. In addition, TMEM16A was co-immunoprecipitated with the inositol 1,4,5-trisphosphate receptor (IP3R) and was activated by IP3R-mediated Ca2+ release. TMEM16A inhibition reduced the IP3R-mediated Ca2+ release induced by cerulein. Furthermore, TMEM16A overexpression activated nuclear factor-κB (NFκB) and increased IL-6 release by increasing intracellular Ca2+. TMEM16A knockdown by shRNAs reduced the cerulein-induced NFκB activation by Ca2+. TMEM16A inhibitors inhibited NFκB activation by decreasing channel activity and reducing TMEM16A protein levels in AR42J cells, and it ameliorated pancreatic damage in cerulein-induced AP mice. This study identifies a novel mechanism underlying the pathogenesis of AP by which IL-6 promotes TMEM16A expression via IL-6R/STAT3 signaling activation, and TMEM16A overexpression increases IL-6 secretion via IP3R/Ca2+/NFκB signaling activation in pancreatic acinar cells. TMEM16A inhibition may be a new potential strategy for treating AP.

摘要

TMEM16ACa2+激活的Cl-通道在胰腺腺泡细胞中表达并参与炎症相关疾病。TMEM16A是否有助于急性胰腺炎（AP）的发病机制尚不清楚。这里,我们发现TMEM16A在胰腺组织中的表达增加与胰腺组织和cerulein诱导的AP小鼠模型血清中的白细胞介素-6(IL-6)水平相关.IL-6处理通过IL-6受体(IL-6R)/信号转导和转录激活因子3(STAT3)信号通路促进AR42J胰腺腺泡细胞中TMEM16A的表达。此外,TMEM16A与肌醇1,4,5-三磷酸受体（IP3R）共免疫沉淀，并被IP3R介导的Ca2释放激活。TMEM16A抑制降低了由cerulein诱导的IP3R介导的Ca2释放。此外，TMEM16A过表达激活核因子-κB(NFκB)并通过增加细胞内Ca2+增加IL-6释放。shRNA敲除TMEM16A降低了ca2诱导的NFκB激活。TMEM16A抑制剂通过降低AR42J细胞中的通道活性和降低TMEM16A蛋白水平来抑制NFκB的活化,并改善了cerulein诱导的AP小鼠的胰腺损伤。本研究确定了AP发病机制的新机制，IL-6通过IL-6R/STAT3信号激活促进TMEM16A表达，TMEM16A过表达通过IP3R/Ca2+/NFκB信号激活增加胰腺腺泡细胞中IL-6的分泌。抑制TMEM16A可能是治疗AP的新的潜在策略。