TRAM1 protects AR42J cells from caerulein-induced acute pancreatitis through ER stress-apoptosis pathway.
Chronic endoplasmic reticulum (ER) stress in pancreatic acinar cells has emerged as a major contributor to the recovery of acute pancreatitis (AP). However, the molecular mechanisms linking AP and ER stress remain not fully understood. In this study, we employed caerulein to induce AP-like inflammation in the AR42J rat pancreatic acinar cells to mimic the AP-like acinar cell injury. Caerulein can activate ER stress in AR42J cells, but the molecular link between AP and ER stress remains to be identified. We here reported that translocating chain-associated membrane protein 1 (TRAM1), an ER-resident multispanning membrane protein, was involved in the onset of AP-like injury on AR42J cells. TRAM1 was significantly elevated in caerulein-treated AR42J cells. Furthermore, we showed that knockdown of TRAM1 led to hyperactivation of 78 kDa glucose-regulated protein precursor (GRP78) and C/EBP homologous protein (CHOP) and the activation of downstream apoptosis pathway. Given the fact that the activation of ER stress played a protection role in AP, the pro-inflammatory mediators TNF-α and IL-6 and the marker of cell injury LDH were also analyzed. We found that depletion of TRAM1 markedly increased the secretion of TNF-α, IL-6, and LDH in the cells. Moreover, flow cytometry indicated that treatment with caerulein induced a significant decrease of apoptotic index and increase of necrosis index in TRAM1-siRNA cells, compared with control groups, as indicated by downregulated expression of cleaved caspase-3, caspase-8, and caspase-9 mRNA expression activity in TRAM1-siRNA cells. These data implicated that TRAM1 might protect AR42J cells against caerulein-induced AP in AR42J cells through alleviating ER stress.
Department of Emergency, Affiliated Hospital of Nantong University, Nantong University, Nantong, Jiangsu Province, China.|Department of Pathogen Biology, Medical College, Nantong University, Nantong, Jiangsu Province, China.
Yongxia
Y
Cai
Department of Emergency, Affiliated Hospital of Nantong University, Nantong University, Nantong, Jiangsu Province, China.
Yanbo
Y
Shen
Department of Emergency, Affiliated Hospital of Nantong University, Nantong University, Nantong, Jiangsu Province, China.
Guangling
G
Xu
Department of Emergency, Affiliated Hospital of Nantong University, Nantong University, Nantong, Jiangsu Province, China.
Ran
R
Tao
Department of Gastroenterology, Affiliated Hospital of Nantong University, Nantong University, Nantong, Jiangsu Province, China.
Weiyan
W
Yuan
Department of Emergency, Affiliated Hospital of Nantong University, Nantong University, Nantong, Jiangsu Province, China. zhongweihuang88@163.com.
Zhongwei
Z
Huang
Department of Pathogen Biology, Medical College, Nantong University, Nantong, Jiangsu Province, China. zdm@ntu.edu.cn.
Dongmei
D
Zhang
D061354:Acinar Cells / Q000473:pathology*; D000818:Animals; D017209:Apoptosis; D002460:Cell Line; D002108:Ceruletide / Q000494:pharmacology; D059865:Endoplasmic Reticulum Stress; D055785:Gene Knockdown Techniques; D008562:Membrane Glycoproteins / Q000378:metabolism / Q000502:physiology*; D026901:Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D016328:NF-kappa B / Q000378:metabolism; D010195:Pancreatitis / Q000473:pathology*; D034622:RNA Interference; D051381:Rats
D008562:Membrane Glycoproteins; D026901:Membrane Transport Proteins; D016328:NF-kappa B; C000614084:TRAM1 protein, rat; D002108:Ceruletide
AR42J cells; Acute pancreatitis; Caerulein; ER stress; TRAM1