Esmolol inhibits inflammation and apoptosis in the intestinal tissue via the overexpression of NF-κB-p65 in the early stage sepsis rats

Abstract

Background/aims: Accumulating evidence reveals esmolol could protect the gut mucosa through the regulation of immune response and inflammation in patients with sepsis. However, its underlying mechanism is not fully understood.

Materials and methods: Diamine oxidase (DAO), intestinal fatty acid-binding protein (I-FABP), interleukin (IL)-6, and IL-10 in the plasma of rats were detected by ELISA assay. Western blotting was utilized to measure the expression levels of NF-kappa B-p65, Bcl-2, and cleaved caspase-3 in the intestinal tissues. The survival analysis was performed in each group.

Results: The plasma levels of DAO and IL-10 levels were increased, whereas that of I-FABP and IL-6 were decreased in the sepsis rats after esmolol treatment, indicating that after the esmolol treatment, the intestinal inflammation and damages were remarkably reduced as compared to those in the normal saline treated sepsis rats. NF-κB-p65 and Bcl-2 were highly expressed, but cleaved caspase-3 showed lower expression in the esmolol treated groups. However, at the same time, we observed contrasting results in the normal saline treated group. Western blotting data indicated that the esmolol treatment inhibited the inflammation and apoptosis in the intestinal tissue due to the overexpression of NF-κB-p65 in the celiac sepsis rats. The survival analysis results indicate that the esmolol infusion should be used in the early stages sepsis rats.

Conclusion: Esmolol can suppress inflammation and apoptosis in the intestinal tissue via the overexpression of NF-kappa B-p65 in the early stage sepsis rats. kappa BEarly-stage use of esmolol might be an ideal treatment method for sepsis.

Figure 1. a–d

ELISA assay shows that esmolol increases DAO and IL-10 concentration and decrease i-FABP and IL-6 concentration in sepsis rat plasmas. a, d) For the DAO concentrations, which in corresponding subgroups of CNS and CE group lower than those in SNS group, but which in CE group higher than those in CNS group. b, c) For the I-FABP or IL-6 concentrations, which in corresponding subgroups of CNS and CE group higher than those in SNS group, but which in CE group lower than those in CNS group. D. For the IL-10 concentrations, which in corresponding subgroups of CNS and CE group higher than those in SNS group, but which in CE group higher than those in CNS group. The asterisk (*) means corresponding subgroups of SNS group compared with those in CNS group and CE group * p<0.05, ** p<0.01. The well number (#) means corresponding subgroups of CNS group compared with those in CE group # p<0.05, ## p<0.01.

Figure 2. a–i

Intestinal tissue pathological changes of sepsis rats. Rats were treated with esmolol for 6h, 12h and 24h, intestinal tissues from SNS, CNS and CE group were stained with HE, inflammatory changes were observed under microscope. No significant pathological changes were found in Sham group (a, d, g). Significant pathological changes of tissue were found in each subgroups of CNS group, which include epithelial cells swell and destroyed, tissue necrogenesis, inflammatory cells infiltrating (b, e, h): b) epithelial cells appear swell, relative less inflammatory cells infiltrating. e, h) epithelial cells appear swelling and destroyed, tissue necrogenesis, inflammatory cells infiltrating. These tissue destroy and inflamatic changes were reduced in each corresponding subgroups of CE group (c, f, i). HE staining 200 ×.

Figure 3

Villus length was quantified in sections of small intestinal. Sepsis-induced villus atrophy is exacerbated in CNS group (HE stain). CNS and CE group rats had markedly shorter villi than SNS group. CNS group had significantly more serious villus atrophy and shorter villi than CE group. Magnification 100x. Villus length was quantified in sections of small intestinal. n=6/group. The asterisk (*) means the villus length in CNS and CE group compared with those in SNS group. ** p<0.01. The well number (#) means subgroups of CNS group compared with corresponding those in. ## p<0.01.

Figure 4. a–d

Survival analysis of rats in SNS, CNS and CE groups. The 6hS, 12hS and 24hS mortalities in CEgroup lower than corresponding those in CNS group. The 12hS and 24hS have higher survival rates than 6hS in CE group, but there is no survival rate difference between 12hS and 24hS in CE group (* p<0.05, ** p<0.01). All sham animals survived.

Figure 5. a, b

Esmolol inhibits apoptosis via NF-kappa B-p65 overexpression. For 24 hours treated subgroups, there is no significant difference of NF-kappa B-p65 and Bcl-2 between CNS and SNS group, but those in CE group has a significant higher expression than SNS group. The level of cleaved caspase-3 has a significantly higher level in CNS group than SNS and CE group (**p<0.01).