Xu Lan-Tao, Xu He-Ling, Fu Ming-Sheng
Department of Gastroenterology, Ruijin Hospital North Affiliated to Shanghai Jiaotong University School of Medicine Shanghai 201801, China.
Soochow University Suzhou, 215006, China.
Int J Clin Exp Pathol. 2015 Aug 1;8(8):9300-6. eCollection 2015.
This study aimed to investigate the role of glucose regulated protein 78 (GRP-78) in the apoptosis of neutrophils in rats with severe acute pancreatitis.
A total of 54 SD male rats were randomly assigned into 2 groups: sham group (n=24) and pancreatitis group (n=30). Severe acute pancreatitis was induced by retrograde cholangiopancreatography injection of sodium taurocholate. Rats were sacrified at 3 h, 6 h and 12 h after injection. In control group, rats received laparotomy, but the pancreates remained intact. The serum amylase was detected at different time points, and flow cytometry was done to detect the apoptosis of neutrophils. Proteins were extracted from neutrophils and subjected to detection of GRP78 and Mcl-1 expression by Western blot assay. HE staining was performed for pathological scoring of the pancreas.
The serum amylase in pancreatitis group increased markedly when compared with control group (P<0.01). In SAP group, the serum amylase increased gradually over time (P<0.01). HE staining showed a lot of inflammatory cells and infiltration of red blood cells and the apoptosis rate of neutrophils reduced gradually (P<0.01). Western blot assay showed the protein expression of GRP-78 and Mcl-1 increased in neutrophils over time.
In rats with SAP, the apoptosis rate of neutrophils reduced over time, which may be associated to the stress induced expression of GRP78 and subsequent activation of Mcl-1 resulting in suppression of neutrphil apoptosis over time.
本研究旨在探讨葡萄糖调节蛋白78(GRP - 78)在重症急性胰腺炎大鼠中性粒细胞凋亡中的作用。
将54只雄性SD大鼠随机分为2组:假手术组(n = 24)和胰腺炎组(n = 30)。通过逆行胆管胰管造影注射牛磺胆酸钠诱导重症急性胰腺炎。在注射后3小时、6小时和12小时处死大鼠。对照组大鼠接受剖腹手术,但胰腺保持完整。在不同时间点检测血清淀粉酶,并通过流式细胞术检测中性粒细胞的凋亡情况。从中性粒细胞中提取蛋白质,通过蛋白质免疫印迹法检测GRP78和Mcl - 1的表达。对胰腺进行苏木精 - 伊红(HE)染色以进行病理评分。
与对照组相比,胰腺炎组血清淀粉酶显著升高(P < 0.01)。在重症急性胰腺炎(SAP)组中,血清淀粉酶随时间逐渐升高(P < 0.01)。HE染色显示有大量炎性细胞以及红细胞浸润,中性粒细胞凋亡率逐渐降低(P < 0.01)。蛋白质免疫印迹法显示中性粒细胞中GRP - 78和Mcl - 1的蛋白质表达随时间增加。
在重症急性胰腺炎大鼠中,中性粒细胞凋亡率随时间降低,这可能与应激诱导的GRP78表达及随后Mcl - 1的激活有关,从而导致中性粒细胞凋亡随时间受到抑制。