Department of Gastroenterology, The First Affiliated Hospital of Guangxi Medical University, Nanning 530021, China; Department of Gastroenterology, The First People's Hospital of Nanning City, The Fifth Affiliated Hospital of Guangxi Medical University, Nanning 530022, China.
Department of Chemotherapy, Affiliated Tumor Hospital of Guangxi Medical University, Nanning 530021, China.
Hepatobiliary Pancreat Dis Int. 2021 Aug;20(4):366-375. doi: 10.1016/j.hbpd.2020.05.004. Epub 2020 Jun 2.
Previous studies have provided conflicting results regarding whether the serum ghrelin concentration can reflect the severity of acute pancreatitis (AP). The present study examined the correlation between the serum ghrelin concentration and AP severity in animal models and investigated whether altered ghrelin expression in pancreatic acinar cells influences IKKβ/NF-κB signaling and pro-inflammatory cytokine production.
Mild or severe AP was induced in rats by intraperitoneal injection of cerulein or retrograde cholangiopancreatic duct injection of sodium taurocholate, respectively. After successful model induction, serum ghrelin, tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6) concentrations were determined by enzyme-linked immunosorbent assay, and IKKβ/NF-κB activation was assessed by immunohistochemistry. Subsequently, stable overexpression or knockdown of ghrelin in AR42J cells was achieved by lentiviral transfection. After transfected cells and control cells were treated with cerulein for 24 h, the TNF-α and IL-1β levels in the supernatants were determined by enzyme-linked immunosorbent assay, and the expression levels of p-p65, IKKβ, and p-IKKβ were detected by Western blotting.
In rat AP models, AP severity was correlated with increased IKKβ/NF-κB activation, pro-inflammatory cytokine production, and ghrelin secretion. The levels of pro-inflammatory cytokines TNF-α and IL-1β as well as IKKβ/NF-κB signaling activity were increased upon knockdown of ghrelin in the AP acinar cell model and decreased with ghrelin overexpression.
Serum ghrelin is related to the severity of AP. Ghrelin may play a protective role in the pathogenesis of AP by inhibiting the pro-inflammatory cytokines and the activation of the IKKβ/NF-κB signaling pathway.
之前的研究对于血清生长激素释放肽（ghrelin）浓度能否反映急性胰腺炎（AP）的严重程度提供了相互矛盾的结果。本研究在动物模型中检查了血清 ghrelin 浓度与 AP 严重程度之间的相关性，并研究了胰腺腺泡细胞中 ghrelin 表达的改变是否会影响 IKKβ/NF-κB 信号通路和促炎细胞因子的产生。
通过腹腔内注射鹅去氧胆酸钠或逆行胰胆管注射牛磺胆酸钠分别诱导大鼠轻度或重度 AP。成功诱导模型后，通过酶联免疫吸附试验测定血清 ghrelin、肿瘤坏死因子-α（TNF-α）和白细胞介素-6（IL-6）浓度，并通过免疫组织化学评估 IKKβ/NF-κB 激活情况。随后，通过慢病毒转染实现 AR42J 细胞中 ghrelin 的稳定过表达或敲低。转染细胞和对照细胞用鹅去氧胆酸钠处理 24 小时后，通过酶联免疫吸附试验测定上清液中 TNF-α和 IL-1β的水平，并通过 Western blot 检测 p-p65、IKKβ和 p-IKKβ的表达水平。
在大鼠 AP 模型中，AP 的严重程度与 IKKβ/NF-κB 激活、促炎细胞因子产生和 ghrelin 分泌增加相关。在 AP 腺泡细胞模型中敲低 ghrelin 会增加促炎细胞因子 TNF-α和 IL-1β以及 IKKβ/NF-κB 信号通路的活性，而过表达 ghrelin 则会降低这些水平。
血清 ghrelin 与 AP 的严重程度有关。Ghrelin 通过抑制促炎细胞因子和 IKKβ/NF-κB 信号通路的激活，在 AP 的发病机制中可能发挥保护作用。
