The Protection And Mechanism Of Shenfu Injection On Acute Liver Injury Of Sepsis Young Rats

Objective:This experiment is aiming to explore the protection and mechanism of Shenfu Injection(SFI) on acute liver injury through the model of sepsis young rats with lipopolysaccharide(LPS). We observe the sepsis young rats the pathological changes of liver, serum levels of liver biochemical indicators(ALT,AST),the expression of TNF-α in liver, the Ach levels in liver tissues, the activities of Ch AT and ACh E in liver tissues by applying SFI, and the correlation between TNF-α with Ach, Ch AT and ACh E.Methods:1. Grouping: Totally 48 SD rats of 4 week-old were randomly divided into three groups: the normal control group(Ctrl group, n=16),the sepsis group(LPS group,n=16),the Shenfu Injection treatment group(SFI group, n=16),each group was further randomly divided into two time points at 6h,12h(n=8 at each group time).2. Model establishing and Drug intervention:The septic model was established by intra-peritoneal injection 10mg/kg LPS. Control Group:The rats were received intra-peritoneal injection of 0.9% saline. LPS Group : The rats were received intra-peritoneal injection of LPS(10mg/kg)and received 0.9% saline(10ml/kg)after1 hour. SFI Group: The rats were received intra-peritoneal injection of Shenfu Injection(10ml/kg) at 1 hour after being intra-peritoneal injection of 10mg/kg LPS.3. Laboratory indexes:The general situation of rats was observed. Rats in each group were euthanized at 6h, 12 h time points. Blood samples were obtained for observing the serum activities of ALT and AST. Liver samples were obtained for measuring the content of Ach, the activities of Ch AT and ACh E, the expression of TNF-α by immunohistochemistry, and the pathological changes of hepatic tissues.Results:1. General situation: Ctrl group rats were in good condition and not found abnormal. LPS group rats got sepsis symptom such as less activity, lethargy, poor response, curled up, shivering, standing hair, fast breathing rate, diarrhea even bleeding. The general situation of SFI group were better than LPS group.2. Compared with Ctrl group, in the LPS group at the same time point the serum levels ALT 、 AST, the expression of TNF-α in liver significantly increase(p <0.01)and the content of Ach, the activities of Ch AT and ACh E of hepatic tissue significantly decrease(p<0.01). The pathological changes of hepatic tissues showed that cellular edema, the inflammatory infiltration and necrosis.3. Compared with LPS group, in the SFI group at the same time point the serum levels ALT 、 AST, the expression of TNF-α in liver significantly decrease(p <0.01)and the content of Ach, the activities of Ch AT of hepatic tissue significantly increase(p<0.01).It could increase the activities of ACh E of hepatic tissue but there was no statistical significance(p > 0.05). The cellular edema, inflammatory infiltration and necrosis of hepatic tissues could be reduced.4. Correlation analysis: the expression of TNF-α was negatively correlated with the content of Ach of hepatic tissues at 6h,12 h in LPS group(r=﹣0.845,﹣0.917,p<0.01),the expression of TNF-α was negatively correlated with the activities of Ch AT in LPS group(r=﹣0.883,﹣0.937,p<0.01).Conclusion:1. Acute liver injury can appear early in sepsis young rats.2. SFI plays a protective effect on acute liver injury of sepsis young rats.3. The mechanism is possibly owing to cholinergic anti-inflammatory pathway through enhancing the activities of Ch AT to increase the levels of Ach, and then inhibiting the expression of TNF-α.