ABSTRACT
Objective To explore a simplified and economical method to isolate the murine primary pancreatic acinar cells.Methods The collagenase and trypsin inhibitor dissolving in DMEM solution were used to digest the murine pancreas,and 4％ BSA dissolving in DMEM solution was used to purify and isolate primary pancreatic acinar cells from pancreas.CCK-8 method was applied to check the ability of pancreatic acinar cells to secret amylase.Results After digestion,shaking in the water bath,resuspension,filtration and precipitation,murine primary pancreatic acinar cells could be obtained within 2 hours.Pancreatic acinar cells in good conditions appeared in clusters,and their basolateral domains were round and devoid of blebs,and the cytoplasm appeared clear.Their apical domain were surrounded by hundreds of zymogen granules which looked darker.The nucleus was located in the basal area of the vesicular region.The basal level of amylase release as a percent of total release from pancreatic acinar cells was around 2.5％ in CCK8-unstimulated group.This rate started to increase after CCK-8 stimulation and reached its peak [(12.83 ± 1.04) ％] at a concentration of 50 pmol/L of CCK-8,but the ratio of the amylase level secreted by the pancreatic acinar cells to the total amylase level displayed a decreasing trend with the increase of CCK-8 concentration.Conclusions This optimized method had the advantage of being fast and simple,low technical difficulty and good repetition.It was a new simplified and cheap method for isolating murine pancreatic acinar cells.
ABSTRACT
Objective To explore the effects of ApoC3 gene on the severity of hypertriglyceridemiainduced acute pancreatitis (AP).Methods ApoC3 transgenetic mice and C57BL/6J mice AP model was induced by cerulein intraperitoneal injection,and ApoC3 transgenetic mice and C57BL/6J mice injected by normal saline solution in equal volume served as control group.Serum triglyceride and cholesterol were detected,and the pathological changes of the pancreas were observed.RT PCR method was used to examine the changes of the inflammatory factor including IL-1β,IL-6,α-SMA and TNF-α mRNA levels,which reflected the severity of the inflammation.Results Serum triglyceride and cholesterol were higher in ApoC3 transgenetic mice than in C57BL/6J mice [(3.434 ± 0.931) mmol/L vs (0.766 ± 0.120) mmol/L,(2.553 ±0.178) mmol/L vs (1.996 ± 0.080) mmol/L],and the differences were statistically different (P ＜ 0.05).The pathological changes of the pancreas were more severe in ApoC3 transgenetic AP mice than in C57BL/6J AP mice,and the IL-1β,IL-6 and α-SMA mRNA levels in the pancreatic tissue were obviously higher in ApoC3 transgenetic AP mice than in C57BL/6J mice (1.72 ± 0.07vs 0.78 ± 0.09,1.58 ± 0.09vs 0.87 ±0.04,0.83 ± 0.05vs 0.44 ± 0.04),and the differences were statistically significant (P ＜ 0.05),while there was no statistical difference on TNF-αmRNA level (0.70 ± 0.09vs 0.65 ± 0.08,P ＞ 0.05).Conclusions ApoC3 gene could aggravate the severity of the inflammation in hypertriglyceridemia-induced AP.