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OBJECTIVE@#To investigate the protective effect of serine hydroxymethyl transferase 2 (SHMT2) against hepatic ischemia-reperfusion injury in mice.@*METHODS@#Sixty C57BL/6 mice were divided equally into sham-operated group, saline adeno-associated virus group (AVV-GFP), and adeno-associated virus silencing group (AAV-SHMT2). The adeno-associated virus and normal saline were injected into the tail vein of the mice 2 weeks before establishment of a 70% ischemia-reperfusion model in the liver. qPCR, Western blotting, immunofluorescence and immunohistochemistry were used to detect the changes of AST/ALT concentration, SHMT2, JNK, NF-κB, caspase-3 and downstream inflammatory factors in the mice, and HE staining was used to observe the pathological damage of the liver tissue in each group; the cell apoptosis in the liver was detected using TUNEL assay.@*RESULTS@#The expression of SHMT2 increased with time after hepatic ischemia-reperfusion and reached the highest level at 24 h (the relative expression was 1.5, < 0.05). At 24 h after hepatic ischemia-reperfusion, the levels of AST/ALT in AAV-SHMT2 group (588/416 U/L) were significantly higher than those in the control group (416/345 U/L) and the empty vector group (387/321 U/L) ( < 0.05). Compared with those in the control group and the empty vector group, the level of SHMT2 was significantly decreased in AAV-SHMT2 group (with a relative expression of 0.24, < 0.05), the levels of p-JNK and p-p65 were significantly increased (relative expression of 0.80 and 0.97, respectively, < 0.05), and the levels TNF-α and IL-1β were consistently elevated (relative expression levels of 1.6 and 1.2, respectively, < 0.05). No significant differences were found in these parameters between the empty vector group and the control group (>0.05).@*CONCLUSIONS@#SHMT2 may alleviate liver cell apoptosis in mice with hepatic ischemia-reperfusion injury by inhibiting the activation of JNK pathway and excessive activation of NF-κB pathway to reduce hepatic damage.
Subject(s)
Animals , Mice , Apoptosis , Liver , Methyltransferases , Mice, Inbred C57BL , NF-kappa B , Reperfusion Injury , SerineABSTRACT
Acute pancreatitis, chronic pancreatitis and pancreatic ductal adenocarcinoma are common pancreatic diseases. Interleukin-6 (IL-6) is a multipotent cytokine, which plays an important role in the differentiation of the severity of pancreatic diseases, prognosis and treatment of pancreatic diseases. This article reviews recent studies on the role of IL-6 in pancreatic diseases.
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ObjectiveTo investigate the effect of triptolide on fibrosis in mice with cerulein-induced chronic pancreatitis (CP) based on the nuclear factor-kappa B (NF-κB)/interleukin-6 (IL-6) signaling pathway. MethodsA total of 15 male C57BL/6 mice were randomly divided into control group (treated with normal saline), cerulein group (model mice with cerulein-induced CP), and triptolide group (induced by cerulean and treated with triptolide), with 5 mice in each group. Samples were collected for detection after 6 weeks. The weight of the pancreas was measured; HE staining, picrosirius red staining, and Masson staining were used to observe pancreatic histomorphology and collagen deposition; ELISA was used to measure the expression of IL-6 in serum; immunohistochemistry was used to measure the expression of alpha-smooth muscle actin (α-SMA) and NF-κB/p65; Western blot was used to measure the expression of NF-κB/p65, IL-6, and α-SMA. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups. ResultsThere were significant differences between the three groups in the weight of the pancreas, IL-6 concentration, pathological score, picrosirius red staining results, Masson staining results, and mean optical density of positive α-SMA signal and positive NF-κB/p65 signal(F=64.87, 15.85, 145.33, 141.80, 121.77, 250.22, and 69.22, all P＜0.001). Compared with the cerulein group, the triptolide group had a significant increase in the weight of the pancreas and significant reductions in the expression of IL-6, pancreatic fibrosis, collagen deposition, and the expression of α-SMA and NF-κB/p65 (all P＜0.05). Western blotting showed that there were significant differences in the expression of NF-κB/p65, IL-6, and α-SMA between the three groups (F=8.86, 6.74, and 16.23, all P＜0.05), and compared with the cerulein group, the triptolide group had significantly lower expression of NF-κB/p65, IL-6, and α-SMA in the pancreatic tissue (all P＜0.05). ConclusionTriptolide alleviates fibrosis in mice with cerulein-induced CP and inhibit the protein expression of NF-κB/p65 and IL-6.
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ObjectiveTo investigate the effect of triptolide on fibrosis in mice with cerulein-induced chronic pancreatitis (CP) based on the nuclear factor-kappa B (NF-κB)/interleukin-6 (IL-6) signaling pathway. MethodsA total of 15 male C57BL/6 mice were randomly divided into control group (treated with normal saline), cerulein group (model mice with cerulein-induced CP), and triptolide group (induced by cerulean and treated with triptolide), with 5 mice in each group. Samples were collected for detection after 6 weeks. The weight of the pancreas was measured; HE staining, picrosirius red staining, and Masson staining were used to observe pancreatic histomorphology and collagen deposition; ELISA was used to measure the expression of IL-6 in serum; immunohistochemistry was used to measure the expression of alpha-smooth muscle actin (α-SMA) and NF-κB/p65; Western blot was used to measure the expression of NF-κB/p65, IL-6, and α-SMA. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups. ResultsThere were significant differences between the three groups in the weight of the pancreas, IL-6 concentration, pathological score, picrosirius red staining results, Masson staining results, and mean optical density of positive α-SMA signal and positive NF-κB/p65 signal(F=64.87, 15.85, 145.33, 141.80, 121.77, 250.22, and 69.22, all P＜0.001). Compared with the cerulein group, the triptolide group had a significant increase in the weight of the pancreas and significant reductions in the expression of IL-6, pancreatic fibrosis, collagen deposition, and the expression of α-SMA and NF-κB/p65 (all P＜0.05). Western blotting showed that there were significant differences in the expression of NF-κB/p65, IL-6, and α-SMA between the three groups (F=8.86, 6.74, and 16.23, all P＜0.05), and compared with the cerulein group, the triptolide group had significantly lower expression of NF-κB/p65, IL-6, and α-SMA in the pancreatic tissue (all P＜0.05). ConclusionTriptolide alleviates fibrosis in mice with cerulein-induced CP and inhibit the protein expression of NF-κB/p65 and IL-6.