Yu Can, Yu Xiao, Zhu Hong-Wei, Li Xia, Huang Li-Hua, Li Zhi-Qiang, Han Duo, Huang Hui
Department of Hepatobiliary and Pancreatic Surgery, Third Xiangya Hospital, Central South University, Changsha, Hunan 410013, P.R. China.
Department of Endocrinology, Third Xiangya Hospital, Central South University, Changsha, Hunan 410013, P.R. China.
Mol Med Rep. 2016 Dec;14(6):5507-5513. doi: 10.3892/mmr.2016.5945. Epub 2016 Nov 15.
High-motility group box protein 1 (HMGB1) has an important role in autophagy; however, its exact role in acute necrotizing pancreatitis (ANP) remains unknown. The present study aimed to investigate the expression pattern of HMGB1 in ANP, and to determine its association with autophagy. Sprague Dawley rats (weight, 350±30 g, n=48) were randomly divided into control (n=12) and experimental (n=36) groups. Experimental rats were retrogradely injected with 5% sodium taurocholate into the biliopancreatic duct to induce ANP. Control rats received an equal amount of saline. Serum amylase levels were used to determine whether the model had been successfully generated. Autophagosomes in pancreatic acinar cells were observed under electron microscopy. The expression levels of HMGB1 and Beclin 1 were detected in pancreatic tissues by western blotting, quantitative polymerase chain reaction and immunohistochemistry. HMGB1 levels were also determined in the serum and in isolated nuclei. The results demonstrated that autophagy was detected at 3 h post‑ANP induction; however, HMGB1 expression remained unaltered during the early stage (0‑6 h; P>0.05). HMGB1 expression was significantly increased at 12 h, and was still increasing at 24 h (P<0.05). Notably, HMGB1 was increased in the nuclei compared with in the cytoplasm at 3‑6 h. Furthermore, serum HMGB1 levels began to increase at 3 h, and reached the highest levels at 24 h in the ANP group. In conclusion, in an ANP model, HMGB1 was initially increased in the nuclei to initiate autophagy. Subsequently, it moved into the cytoplasm, where it interacted with Beclin 1 to enhance autophagy, and HMGB1 was released into the blood, leading to the deterioration of ANP.
高迁移率族蛋白B1(HMGB1)在自噬中发挥重要作用;然而,其在急性坏死性胰腺炎(ANP)中的确切作用尚不清楚。本研究旨在探究HMGB1在ANP中的表达模式,并确定其与自噬的关联。将体重为350±30 g的Sprague Dawley大鼠(n = 48)随机分为对照组(n = 12)和实验组(n = 36)。对实验组大鼠经胰胆管逆行注射5%牛磺胆酸钠以诱导ANP。对照组大鼠注射等量的生理盐水。采用血清淀粉酶水平来确定模型是否成功构建。通过电子显微镜观察胰腺腺泡细胞中的自噬体。采用蛋白质免疫印迹法、定量聚合酶链反应和免疫组织化学法检测胰腺组织中HMGB1和Beclin 1的表达水平。还测定了血清和分离细胞核中的HMGB1水平。结果表明,在ANP诱导后3小时检测到自噬;然而,在早期阶段(0 - 6小时)HMGB1表达未发生改变(P>0.05)。HMGB1表达在12小时时显著增加,并在24小时时仍在升高(P<0.05)。值得注意的是,在3 - 6小时时,细胞核中的HMGB1较细胞质中增加。此外,ANP组血清HMGB1水平在3小时时开始升高,并在24小时时达到最高水平。总之,在ANP模型中,HMGB1最初在细胞核中增加以启动自噬。随后,它转移至细胞质中,在那里与Beclin 1相互作用以增强自噬,并且HMGB1释放到血液中,导致ANP病情恶化。