Tang Xiping, Tang Guodu, Liang Zhihai, Qin Mengbin, Fang Chunyun, Zhang Luyi
From the *Department of Gastroenterology, Second Affiliated Hospital of Guangxi Medical University; and †Department of Gastroenterology, First Affiliated Hospital of Guangxi Medical University, Nanning, China.
Pancreas. 2017 Nov/Dec;46(10):1305-1313. doi: 10.1097/MPA.0000000000000946.
The study investigated the effects of endogenous targeted inhibition of ghrelin gene on inflammation and calcium pathway in an in vitro pancreatic acinar cell model of acute pancreatitis.
Lentiviral expression vector against ghrelin gene was constructed and transfected into AR42J cells. The mRNA and protein expression of each gene were detected by reverse transcription polymerase chain reaction, Western blotting, or enzyme-linked immunosorbent assay. The concentration of intracellular calcium ([Ca]i) was determined by calcium fluorescence mark probe combined with laser scanning confocal microscopy.
Compared with the control group, cerulein could upregulate mRNA and protein expression of inflammatory factors, calcium pathway, ghrelin, and [Ca]i. mRNA and protein expression of inflammatory factors increased significantly in cells transfected with ghrelin miRNA compared with the other groups. Intracellular calcium and expression of some calcium pathway proteins decreased significantly in cells transfected with ghrelin miRNA compared with the other groups.
Targeted inhibition of ghrelin gene in pancreatic acinar cells of acute pancreatitis can upregulate the expression of the intracellular inflammatory factors and alleviate the intracellular calcium overload.
本研究在急性胰腺炎体外胰腺腺泡细胞模型中，探究内源性靶向抑制胃饥饿素基因对炎症和钙信号通路的影响。
构建针对胃饥饿素基因的慢病毒表达载体并转染至AR42J细胞。通过逆转录聚合酶链反应、蛋白质印迹法或酶联免疫吸附测定法检测各基因的mRNA和蛋白质表达。采用钙荧光标记探针结合激光扫描共聚焦显微镜测定细胞内钙浓度（[Ca]i）。
与对照组相比，雨蛙肽可上调炎症因子、钙信号通路、胃饥饿素及[Ca]i的mRNA和蛋白质表达。与其他组相比，转染胃饥饿素微小RNA的细胞中炎症因子的mRNA和蛋白质表达显著增加。与其他组相比，转染胃饥饿素微小RNA的细胞中细胞内钙及一些钙信号通路蛋白的表达显著降低。
在急性胰腺炎胰腺腺泡细胞中靶向抑制胃饥饿素基因可上调细胞内炎症因子的表达并减轻细胞内钙超载。
