Effect of unfractionated heparin on the expression of heme oxygenase-1 in intestinal mucosa of mice with sepsis / 中华危重病急救医学

ABSTRACT

Objective To investigate the effect of unfractionated heparin (UFH) on the expression of heme oxygenase-1 (HO-1) in intestinal mucosa of mice with sepsis.Methods Thirty-six male C57BL/6J mice were randomly divided into sham group,cecal ligation and puncture (CLP) group and UHF group,n =12 in each group.Model of intestinal injury in sepsis was induced by CLP.In sham group,the mice were exposed without ligation of cecum.In UFH group,the mice were treated intravenously with 8 U of UFH via the tail vein half an hour before the operation and 12 hours after the surgery respectively.Six mice in each group were randomly chosen at 4 hours and 24 hours after operation to collect inferior vena venous blood samples and terminalileum tissues.The serum levels of interleukins (IL-1 β,IL-6),and tumor necrosis factor-α (TNF-α) were determined by enzyme linked immunosorbent assay (ELISA).The serum level of D-lactate was determined by colorimetry.Pathological changes of ileum tissue and Chiu score were observed after hematoxylin eosin (HE) staining.The HO-1 expression was detected immunohistochemically.Results In sham group,no significant changes in the serum levels of IL-1 β,IL-6,TNF-α and D-lactate were observed.Twenty-four hours after the operation,the structure of intestinal mucosa was basically normal without obvious pathology change and no HO-1 positive cells were found.The serum levels of IL-1 β,IL-6,TNF-α,and D-lactate in CLP group were gradually increased,and they were significantly increased as compared with sham group [IL-1 β (ng/L)40.87±2.88 vs.22.60±2.05 at 4 hours,113.73±3.96 vs.22.07±2.74 at 24 hours;IL-6 (ng/L)63.89±3.26 vs.44.89±3.38 at 4 hours,176.56±5.45 vs.45.76±4.02 at 24 hours;TNF-α (ng/L)194.62± 14.13 vs.152.05±6.22 at 4 hours,599.62± 10.20 vs.155.90± 14.18 at 24 hours;D-lactate (mmol/L)0.24± 0.02 vs.0.19 ± 0.01 at 4 hours,0.33 ± 0.04 vs.0.20 ± 0.02 at 24 hours,all P ＜ 0.05].Twenty-four hours after the operation,edema and inflammation in ileal mucosa,intestinal villi structural damage were observed,the Chiu score was significantly higher than those in the sham group [4.5 (3.0-5.0) vs.0 (0-1.0),P ＜ 0.05],and a small amount of HO-1 positive cells were localized in the intestinal mucosa.Compared with CLP group,the serum levels of IL-1 β,IL-6,TNF-α,and D-lactate of UFH group were significantly decreased [IL-1 β (ng/L)31.53 ± 2.90 vs.40.87 ± 2.88 at 4 hours,61.13 ± 2.80 vs.113.73 ± 3.96 at 24 hours;IL-6 (ng/L)51.16 ± 5.68 vs.63.89 ± 3.26 at 4 hours,81.16 ± 4.54 vs.176.56 ± 5.45 at 24 hours;TNF-α (ng/L)171.76± 5.60 vs.194.62± 14.13 at 4 hours,328.48 ± 10.79 vs.599.62± 10.20 at 24 hours;D-lactate (mmol/L)0.21 ±0.01 vs.0.24±0.02 at 4 hours,0.24±0.02 vs.0.33±0.04 at 24 hours,all P ＜ 0.05].Twenty-four hours after the operation,intestinal injury was ameliorated,the Chiu score was significantly lower [1.5 (1.0-5.0) vs.4.5 (3.0-5.0),P ＜ 0.05],and HO-1 positive cells in the intestinal mucosa was remarkably increased.Conclusion UFH can enhance the expression of HO-1 in intestinal mucosa,reduce the release of inflammatory factors,ameliorate the intestinal inflammatory response,and thus play a protective role in intestinal tissue in mice with sepsis.