ABSTRACT
Objective To observe the apoptosis or oncosis of pancreatic acinar cells of different severity of acute pancreatitis (AP) and the release level of enzymes in vitro, and to investigate the relationship between them. Methods Two-step enzymatic digestion method was used to separate pancreatic acinar cells into 4 groups. 0. 1 μg/ml of the caerulein was added in the AP group. Caerulein and LPS (bacterial lipopolysaccharide, 10 mg/L) were added in LPS group. Caerulein and OCT (octreotide, 100 ng/ml) were added in OCT group. Medium was added in the control group. AO (acridine orange) and EB (ethidium bromide) double staining method was used to detect the incidence of apoptosis or oncosis of acinar cell. The release of intracellular enzyme was detected by measuring the concentrations of amylase and LDH in cell culture media by colorimetry method. Results The apoptosis index was 2.2 + 0.4, 6.4 ± 0.6, 4.6 + 0.4, 11.2 +1.2 in the control group, AP group, LPS group, OCT group; while the oncosis index was 3.0 +0.4, 17.2 ±1.6, 23.0 ± 2.2, 12.8 ± 1.4 in the control group, AP group, LPS group, OCT group; the release of LDH was (2180 ±240), (8060 ±930), (9460 +920), (6860 ±740) U/dl, the level of amylase was (1750 ± 190),(3820 ±460), (4420 ±480), (2260 ±260)U/L. All the values in the experiment groups were significantly higher than that in control group ( P ＜ 0.05 ). The oncosis index, LDH, amylase in LPS group was significantly higher than that in AP group ( P ＜ 0.05 ), but the apoptosis index in LPS group was significantly lower than that in AP group ( P ＜ 0.05 ). The apoptosis index in OCT group was significantly higher than that in AP group ( P ＜ 0. 05 ), but the oncosis index, LDH, amylase was significantly lower than that in AP group ( P ＜ 0. 05 ).Conclusions Induction of apoptosis and reduction of oncosis in AP pancreatic acinar cells can reduce the release of enzyme in acinar cells.
ABSTRACT
Objective To study the relationship between the oncosis of pancreatic acinar cells and activa-tion of macrophage in rat model of acute pancreatitis (AP). Methods The pancreatic acinar cells were isolated by two-step enzyme digestion, and then they were divided into control group, AP group and test group. Pancreatic acinar cells were cultured with caerulein in AP group, with caerulein and endothelin in test group, and with culture medium in control group. The oncesis rate of the pancreatic acinar cells was detected after acridine orange and ethidium bromide fluorescent staining. The supernatant was collected to detect the release of amylase and lactate dehydrogenase (LDH). The macrophages were cultured with 1 ml of supematant for 6 hours, and then the protein level of tumor necrosis factor-α (TNF-α) was measured by ELISA. Results Few oncotic pancreatic acinar cells were observed in the control group, and the levels of amylase and LDH secreted by pancreatic acinar cells and TNF-α secreted by macrophage were (1175±165)kU/L, (846±118)U/L and (36±5)μg/L, respectively. Oncotic pancreatic acinar cells were observed in AP group, and the levels of amylase, LDH and TNF-α were (7130±680) kU/L, (4262±626) U/L and (155±18) μg/L, respectively, which were significantly higher than those in control group (t = 5.184, 4.277, 3.665, P < 0.05). The levels of amylase, LDH and TNF-α were even higher in test group, and they were (9240±1177) kU/L, (6937±893)U/L and (268±35)μg/L, respectively, which were significantly higher than those in AP group (t = 2.251, 2.825, 2.843, P < 0.05). Conclusions The release of amylase was changed as the oncosis of pancreatic acinar cells occurred. The secretion of TNF-α was along with the degree of oncosis of pancreatic acinar cells. The results of the study indicate that a relationship exists among the inflammatory response of macrophage, the release of contents of pancreatic acinar cells and the oncosis of the pancreatic acinar cells.