Therapeutic effect of hesperidin on severe acute pancreatitis in rats and its mechanism / 中华危重病急救医学

ABSTRACT

Objective To investigate the protective effect of hesperidin on severe acute pancreatitis (SAP) in rats and its related mechanism.Methods Sixty male Sprague-Dawley (SD) rats were randomly divided into five groups (n = 12 in each group) sham group, SAP model group, dexamethasone group (5 mg/kg), low and high dose of hesperidin groups (10 mg/kg and 20 mg/kg). SAP rats were administered a retrograde infusion of 3.5％ sodium taurocholate solution into the biliopancreatic duct after laparotomy. Sham rats were administered with equivalent saline. The treatment was intravenously injected 5 minutes after operation through femoral vein. After 24 hours, the survival of animals was observed, the level of serum amylase, the volume of ascites and the relative specific gravity of the pancreas were measured; the pathological changes of pancreatic tissue were observed by Hematoxylin-eosin (HE) staining; the levels of serum and pancreatic tissue interleukin (IL-1β, IL-6) and tumor necrosis factor-α (TNF-α) were detected by enzyme linked immunosorbent assay (ELISA); the expression of Toll-like receptor 4 (TLR4), the phosphorylation of IL-1 receptor associated kinase (IRAK1) and nuclear factor-κB (NF-κB) were detected by Western Blot.Results Compared with SAP model group, the 24-hour survival rate were increased in low and high dose of hesperidin groups (83.3％, 100％ vs. 58.3％), the volume of ascites were reduced (mL 7.36±0.91, 6.10±1.02 vs. 13.82±2.06), the levels of serum amylase were reduced (U/L 1081.48±78.23, 1048.58±49.97 vs. 1990.37±127.27), the relative specific gravity of the pancreas were reduced [(7.52±1.02)％, (5.59±0.96)％ vs. (11.22±0.96)％], and the pathological damage of pancreatic tissue were reduced; the levels of serum and pancreatic tissue inflammatory factors were reduced in high dose hesperidin group [serum IL-1β (ng/L) 68.08±10.49 vs. 130.30±23.35, IL-6 (ng/L) 63.88±10.47 vs. 158.41±21.38, TNF-α(ng/L) 10.42±1.49 vs. 18.16±2.01; pancreas IL-1β (pg/μg) 13.87±1.84 vs. 20.08±1.66, IL-6 (pg/μg) 21.90±3.12vs. 38.13±3.57, TNF-α (pg/μg) 1.88±0.20 vs. 4.26±0.58]; the expression of TLR4, and the phosphorylation levels of IRAK1 and NF-κB were decreased in low and high dose of hesperidin groups (the sham operation group was 100, TLR4/β-actin 91.9±15.6, 83.7±11.2 vs. 168.5±9.0, p-IRAK1/IRAK1 117.4±7.6, 104.7±11.5 vs. 173.5±15.8, p-NF-κB p65/NF-κB p65 119.9±9.3, 105.8±12.6 vs. 174.1±13.0), with statistically significant differences (allP < 0.05). The effects of dexamethasone were similar to that of high dose of hesperidin.Conclusions Hesperidin could significantly protect SAP rats, and this protection was related to the inhibition of TLR4/IRAK1/NF-κB signaling pathway, and to the reduction of pro-inflammatory cytokine expressions. The effect of high dose hesperidin (20 mg/kg) was more significant.