Therapeutic effects of Saussurea involucrate injection in severe acute pancreatitis in rats

Abstract

Objective To investigate the therapeutic effects of Saussurea involucrate injection in severe acute pancreatitis (SAP) in rats. Methods A total of 80 healthy Sprague-Dawley rats were divided into eight groups: SAP group (three hours、48 hours), Saussurea involucrate treated group (three hours、48 hours), ulinastatin control group (three hours、48 hours) and sham operation group (three hours、48 hours), 10 rats in each group. After modeling, the rats of SAP group were regularly feeded and the rats of other three group were treated with Saussurea involucrate injection (1.04 mL/kg) intraperitoneal injection, ulinastatin 10 000 U/L tail vein injection, and saline femoral vein injection, respectively and injected every 12 hours. At three hours and 48 hours after treated, blood and pancreatic tissue samples were obtained.The mortality rate, serum amylase level and pathological changes of the pancreas of each group were observed. Serum tumor necrosis factor alpha (TNF-α), interleukin (IL)-6 and IL-10 levels were detected by enzyme linked immunosorbent assay (ELISA). The content of malondialdehyde (MDA) in pancreatic tissues was determined by chemical colorimetry. The level of TNF-α mRNA, IL-6 mRNA and IL-10 mRNA in pancreatic tissues were measured with reverse trascription-polymerase chain reaction (RT-PCR). The activity of nuclear factor kappa B p65 (NF-κB p65) in the pancreatic tissue was tested by immunohistochemistry. Single factor analysis of variance was used to compare multiple groups, and the least significant difference (LSD) method was used in the multiple comparisons between groups. Fisher′s exact probability method was performed for rates comparison. Results At 48 hours, there was no statistically significant difference in mortality rate among Saussurea involucrate treated group, SAP group and ulinastatin groups (all P>0.05). At 48 hours, the histopathology score (8.13±0.64), levels of serum amylase ((2 597.0±214.0) U/L), TNF-α ((254.4±11.6) ng/L), IL-6 ((441.4±14.6) ng/L), levels of pancreatic tissues MDA ((311.0±10.6) mmol/L), TNF-α mRNA(2.04±0.08), IL-6 mRNA(1.77±0.04)and activity of NF-κB p65 ((25.90±2.90)%) of Saussurea involucrate treated group were all lower than those of SAP group (11.40±0.89, (4 780.0±101.0) U/L, (396.0±7.4) ng/L, (664.4±7.6) ng/L, (418.0±10.6) mmol/L, 2.94±0.03, 2.63±0.08 and (51.60±5.27)%; however level of serum IL-10 ((133.5±6.9) ng/L vs (95.1±5.2) ng/L) and IL-10 mRNA of the pancreatic tissue (1.38±0.06 vs 0.85±0.03) significantly increased (F=253.07、441.63、489.40、2 465.00、196.65、477.89、562.79、131.70、560.18、570.04, all P 0.05). Conclusion Saussurea involucrate injection can eliminate oxygen free radicals and prevent to xidation, inhibit NF-κB activation, regulate synthesis and release of cytokines, and alleviate pancreatic injury in SAP rats, but it can not decrease mortality. Key words: Saussurea involucrate injection; Ulinastatin; Severe acute pancreatitis