The role of granzyme B-perforin in selectively inducing apoptosis of macrophages in pancreatitis based on KEGG pathway enrichment analysis
KONG Hongru1, YU Haohui2, CHEN Geer2, CHEN Zimiao3, DAI Shengjie1, SUN Xuecheng4, JIN Yuepeng1, YANG Wenjun1, SUN Hongwei1.
1.Department of Hepatopancreatobiliary Surgery, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325015,China; 2.The First School of Medicine, School of Information and Engineering, Wenzhou Medical University,Wenzhou 325035, China; 3.Department of Endocrinology, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325015, China; 4.Department of Gastroenterology, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325015, China
KONG Hongru,YU Haohui,CHEN Geer, et al. The role of granzyme B-perforin in selectively inducing apoptosis of macrophages in pancreatitis based on KEGG pathway enrichment analysis[J]. JOURNAL OF WEZHOU MEDICAL UNIVERSITY, 2022, 52(12): 972-980.
Abstract：Objective: To investigate the effect of granzyme B (GRB)-perforin (PFP) on macrophage apoptosis during selective induction of pancreatitis. Methods: Human mononuclear macrophage cell line (TPH-1) was stimulated by tumor necrosis factor-α (TNF-α) to simulate the macrophage microenvironment during acute pancreatitis. The optimum concentrations of TNF-α, PFP and GRB were screened by CCK8 test. Apoptosis was detected by flow cytometry in control group, TNF-α group, PFP+GRB group and TNF-α+PFP+GRB group. The cell samples were divided into three groups: normal cell group (NC group), TNF-α stimulation group (Treat1
group) and TNF-α+PFP+GRB stimulation group (Treat2 group). Three groups of cell RNA were extracted for transcriptomic analysis. Gene expression was detected by principal component analysis, differential gene analysis and enrichment analysis. Seven differentially expressed genes (RAB37, GPA33, USH1C, SYTL1, MSRB3,
GPER1, CD1B) were selected in the “cytokine - cytokine receptor interaction pathway” to verify the accuracy of RNA-seq by fluorescence quantitative PCR. Results: Compared to NC group and Treat1 group, the apoptosis rate of Treat2 group was significantly higher. The difference between NC group and Treat1 group and Treat2
group was obvious, with more significant genes, while the difference between Treat1 group and Treat2 group was unconspicuous, but the change of Treat2 group was greater than that of Treat1 group in NC group. Enrichment analysis showed that the differential genes were mainly concentrated in the “cytokine - cytokine receptor interaction pathway”. The results of fluorescence quantitative PCR and RNA-seq were highly consistent. Conclusion:GRB-PFP selectively induces apoptosis of macrophages in pancreatitis and acts as a “cytokine - cytokine receptor interaction pathway” to down-regulate the expression of cytokines and reduces the inflammatory response.
