Therapeutic effects of cFLIP gene silencing on acute necrotizing pancreatitis rats and its mechanism
10.3760/cma.j.issn.1674-1935.2017.06.005
- VernacularTitle:cFLIP基因沉默对急性坏死性胰腺炎大鼠的治疗作用及其机制
- Author:
Baozhi WANG
1
;
Jun WANG
;
Heping PENG
;
Yu DING
;
Xuzhong HU
Author Information
1. 广州医科大学附属第二医院普通外科
- Keywords:
Pancreatitis,acute necrotizing;
RNA,small interfering;
Apoptosis;
cFLIP genes
- From:
Chinese Journal of Pancreatology
2017;17(6):375-379
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the therapeutic effects of cFLIP-siRNA tail venous injection on acute necrotizing pancre.atitis (ANP) rats,and explore the related mechanism.Methods Three pairs of cFLIP siRNAs (cFLIP-siRNA) and negative control(siRNA-NC) were designed and synthesized.The most effective siRNA to inhibit the expression of cFLIP gene was selected for the experiment.Thirty SD rats were treated with retrograde pancreaticobiliary duct injection of 5％ taurocholic acid sodium salt solution to establish ANP rat model.They were randomly divided into ANP group,siRNA-NC group and cFLIP-siRNA group with l0 rats in each group using random number method.The same volume of normal saline,siRNA-NC and cFLIP-siRNA were injected into tail vein,respectively.The rats were sacrificed after 24 h and the blood samples were colelcted.The levels of serum amylase and endotoxin were measured.The pancreatic tissues were collected and routine pathological examination was performed.The expressions of cFLIP-S and caspase-8 protein in pancreatic tissues were detected by immunohistochemistry and Western blotting.Results The levels of serum amylase in ANP group,siRNA-NC group and cFLIP-siRNA group were (1 286 ± 209),(1 297 ± 305) and (552± 256)U/L,the level of serum endotoxin were (136± 32),(128± 56) and (46± 23)ng/L,respectively.The pancreatic pathological scores were (4.97 ± 1.16),(4.92 ± 2.03) and (1.67 ± 1.98).The expression level of cFLIP-S protein were (8.56 ± 2.72),(9.12 ± 2.45) and (3.82 ± 1.46),respectively,which were significantly lower in cFLIP-siRNA group than in ANP group and siRNA-NC group (P ＜ 0.05),and the difference was statistically significant (P ＜ 0.05).The expression level of caspase-8 protein in the three groups were (2.25 ± 1.24),(2.41 ± 1.14) and (5.56 ± 1.79),respectively,which were significantly higher in cFLIP-siRNA group than in ANP group and siRNA-NC group,and the difference was statistically significant (P ＜0.05).However,there was no significant difference between ANP group and siRNA-NC group.Conclusions Inhibition of cFLIP gene expression can reduce pancreatic injury in ANP rats.The possible mechanism was that the up-regulation of caspase-8 can inhibit pancreatic cell necrosis and promote apoptosis.
