The cell culture medium supernatants were collected at 2, 4, 8, 12 and 24 hours respectively, and the amylase, lipase and tumor necrosis factor were detected by enzyme-linked immunosorbent assay (ELISA)- α( TNF- α)、 The expression of interleukin-6 (IL-6) was changed.
Results amylase, lipase and TNF in rain frog element group and rain frog element + lipopolysaccharide group at each time point- α、 IL-6 was higher than that in the control group, and at most time points, the level of rain frog element + lipopolysaccharide group was higher than that in rain frog element group, and the up regulation time was the same or earlier (P & lt; 0.05)。 After 24 hours of stimulation, the apoptosis rate of cerulein Group [(19.2 ± 0.9)%] was significantly higher than that of cerulein + lipopolysaccharide Group [(10.2 ± 0.5)%, P & lt; 0.05]。 The necrosis rate of cerulein Group [(5.8 ± 0.3)%] was significantly lower than that of cerulein + lipopolysaccharide Group [(13.6 ± 0.7)%, P & lt; 0.05]。 The necrosis / apoptosis ratio in SAP group was 5 times higher than that in map group.
Each group was divided into 3 subgroups (n = 10).
Objective to investigate glycogen synthase kinase 3 β( GSK3 β) Expression and role in the pathogenesis of mild / severe acute pancreatitis methods the cell models of mild acute pancreatitis (map) and severe acute pancreatitis (SAP) were constructed by single cerulein and cerulein combined with lipopolysaccharide stimulation.
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The activity of superoxide dismutase (SOD) and the level of malondialdehyde (MDA) in fresh liver were detected.
Objective To observe the effect of hydrogen saturated normal saline (HRS) on mitogen activated protein kinases (MAPKs) pathway activated by reactive oxygen free radicals (ROS) in the liver of acute necrotizing pancreatitis (ANP) rats, and to explore its protective effect on the liver of ANP rats.
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GSK3 β The mRNA level of in the rain frog element group was down regulated for 2 hours [the relative expression was (0.5 ± 0.1) times] and increased significantly at 8 hours [the relative expression was (3.0 ± 1.1) times, P & lt; 0.05], and then decreased gradually; In cerulein + lipopolysaccharide group, it was slowly up-regulated after 8 hours [the relative expression was (1.8 ± 0.6) times, P & lt; 0.05], and reached the peak at 24h [the relative expression was (3.6 ± 0.3) times, P & lt; 0.05]。 GSK3 β And caspase-3 mRNA levels increased, but the difference was not statistically significant (P & gt; 0.05)。 Conclusion map and SAP cell models stimulated by cerulein alone and cerulein combined with lipopolysaccharide are feasible; GSK3 β It may play a role in map and SAP by promoting acinar cell apoptosis.
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Serum amylase (Amy), lipase (LIP), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected.
Hrs group was supplemented with hrs6ml / kg in caudal vein after operation, and hrs20ml / kg was injected subcutaneously.
In so group, only the pancreas and duodenum were turned open and the abdomen was closed.
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The apoptosis rate and necrosis rate were detected by flow cytometry, and GSK3 was detected by real-time fluorescence quantitative polymerase chain reaction (FQ-PCR) β And the expression of Caspase-3, – 8, – 9 mRNA.
ANP model was established by retrograde injection of 5% sodium taurocholate into biliopancreatic duct.
Fixed sections of pancreatic head and liver tissue were taken to observe the pathological changes and score or grade.
Methods 90 male Wistar rats were randomly divided into sham operation (so) group, ANP group and HRS group.
The other groups were given the same dose of normal saline.
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The expressions of phosphorylated c-Jun N-terminal kinase (p-JNK), p-p38 and phosphorylated extracellular signal regulated kinase (p-ERK) were detected by immunohistochemistry.
Results serum Amy (U / L) (5385 ± 572 vs 1146 ± 121), lip (U / L) (1924.9 ± 105.4 vs 86.5 ± 8.9), ALT (U / L) (156.9 ± 15.8 vs 56.1 ± 5.4), AST (U / L) (448.5 ± 53.0 vs 124.6 ± 14.7) and pathological score of pancreas and liver [(13.79 ± 1.36) vs (0.38 ± 0.53)] in ANP rats were significantly higher than those in so group at each time point (P & lt; 0.05); Amy (U / L) (5567 ± 512 vs 5385 ± 572) and lip (U / L) (1856.8 ± 149.3 vs 1924.9 ± 105.4) in HRS group were lower than those in ANP group, but the difference was not statistically significant (P & gt; 0.05)。 ALT (U / L) (134.8 ± 11.8 vs 156.9 ± 15.8), AST (U / L) (326.6 ± 20.5 vs 448.5 ± 53.0) and pathological grade of liver (17.1 vs 23.1) in HRS group were significantly lower than those in ANP group (P & lt; 0.05); The activity of SOD (U / mg) (127.2 ± 11.6 vs 267.4 ± 33.2) in ANP group was significantly lower than that in so group, and the level of MDA (nmol / mg) (12.6 ± 3.1 vs 3.9 ± 1.7) was significantly higher than that in so group (P & lt; 0.05), the activity of SOD (U / mg) (177.4 ± 12.5 vs 127.2 ± 11.6) in HRS group was significantly higher than that in ANP group, and the content of MDA (nmol / mg) (9.7 ± 2.3 vs 12.6 ± 3.1) was significantly lower than that in ANP group (P & lt; 0.05)。 The levels of p-JNK (121.40 ± 10.70 vs 2.60 ± 0.45), p-p38 (63.1 ± 6.9 vs 16.2 ± 1.7) and p-ERK (7.10 ± 1.40 vs 0.90 ± 0.04) in ANP group were significantly higher than those in so group (P & lt; 0.05), the levels of p-JNK (51.30 ± 9.70 vs 121.40 ± 10.70) and p-p38 (16.2 ± 1.7 vs 63.1 ± 7.0) in HRS group were significantly lower than those in ANP group (P & lt; 0.05), but there was no significant difference in the expression of p-ERK (6.70 ± 1.30 vs 7.10 ± 1.40) (P & gt; 0.05)。 Conclusion hrs can effectively reduce the oxidative stress in the liver of ANP rats, reduce the activation of JNK and p38 pathway by ROS, and protect the liver of ANP rats..
