The mechanism of miR-494 negatively regulating ROCK1 and PTEN in inhibiting apoptosis of acute pancreatitis cells
10.3760/cma.j.issn.1671-0282.2021.10.010
- VernacularTitle:miR-494负调控ROCK1、PTEN抑制急性胰腺炎细胞凋亡的机制研究
- Author:
Hui FENG
1
;
Jiangli SUN
;
Zequn NIU
;
Liming WANG
;
Honghong PEI
;
Longfei PAN
Author Information
1. 西安交通大学第二附属医院急诊科, 710004
- Keywords:
Acute pancreatitis;
Apoptosis;
miR-494;
Pro-apoptotic protein
- From:
Chinese Journal of Emergency Medicine
2021;30(10):1210-1215
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the mechanism of miR-494 negatively regulating ROCK1 and PTEN in inhibiting apoptosis of pancreatic cells and participating in the occurrence and development of acute pancreatitis.Methods:Pancreatic acinar cells AR42J from rats were treated by caerulein, and then the levels of amylase, tumor necrosis factor alpha (TNF-α), interleukin 1 (IL-1) and IL-6 in the supernatant of cell culture were detected by ELISA to verify the cell model of acute pancreatitis. RT-PCR was used to detect the expression of miR-494 in normal AR42J cells (control group) and acute pancreatitis cell model (model group). Flow cytometry was used to detect the apoptosis of the control group, negative control miRNA transfected acute pancreatitis cell model (negative control group) and miR-494 transfected acute pancreatitis cell model (miR-494 transfection group). Western blot was used to detect the expression of ROCK1 and PTEN in the control group, negative control group and miR-494 transfection group.Results:The levels of amylase, TNF-α, IL-1 and IL-6 in the supernatant of AR42J cells treated with caerulein for 8 h and 12 h were significantly higher than those at 0 h and the control group ( P<0.05), indicating that the model was successfully constructed. The expression levels of miR-494 at 8 h, 12 h and 24 h after the establishment of acute pancreatitis cell model were significantly higher than those at 4 h and the control group ( P < 0.05). The apoptosis rate of the model group was significantly higher than that of the control group ( P<0.05), and the apoptosis rate of the miR-494 transfection group was significantly lower than that of the model group ( P<0.05). The expression levels of ROCK1 and PTEN in the miR-494 transfection group were significantly lower than those in the model group and negative control group ( P<0.05). Conclusions:When acute pancreatitis occurs, overexpression of miR-494 can inhibit the expression of pro-apoptotic protein, thus inhibiting the apoptosis of pancreatic acinar cells and promoting the development of acute pancreatitis.
