| Background Sepsis is the Systemic inflammatory response due to infection, it is a hot research point in the field of critical care medicine,acute liver injury is an important complication of sepsis and impact on the prognosis. Inflammatory cytokine storm is one of the main pathogenesis of acute liver injury. Cholinergic anti-Inflammatory pathway(CAP) is the pathway to against systemic inflammatory response by cholinergic neurotransmitters regulate,nicotinic acetylcholine receptor ?7subunit(?7n ACh R) is the main regulator of inflammatory cytokine production.Currently ?7n ACh R is gradually becoming the target of antiinflammatory and Having a "switch" action on inflammatory,GTS-21 is the specific agonists of ?7n ACh R, and play an role of anti-inflammatory in Alzheimer's,arthritis, and many other immune inflammatory diseases, but now there are few reports about whether it could protect acute liver injury in mice with LPS.Objective First,to observed the express of ?7n ACh R and IL-18, IL-1?,HMGB1 in acute liver injury of mice model by LPS,explore thepathogenesis of ?7n ACh Ron acute liver injury with LPS; then observed the affect of GTS-21 on the express of ?7n ACh R and Inflammatory cytokines in acute liver injury of mice and explore the protective effect and anti-inflammatory mechanism of GTS-21 in ALI.Methods Experiment 1 study on the pathogenesis of ?7n ACh R in acute liver injury by LPS60 male C57BL/6 mice were randomly divided into normal control group(n = 10); acute liver injury model group(LPS 30mg/kg, n = 50),adaptive feeding one week,the model group were given LPS 30mg/kg by intraperitoneal injection, The mice were sacrificed after modeling 1h, 3h,6h, 12 h, 24 h, normal control group intraperitoneal;normal control group intraperitonealled with an equal volume of LPS co-solvent(0.9%saline),The mice were sacrificed after modeling 1h. Each group selected8 samples randomly,then observe gross morphology of the live,Blood samples were collected and tested alanine aminotransferas(ALT),aspartate aminotransferase(AST) levels,used the ELISA to assay expression of IL-18, IL-1?, HMGB1 in liver tissue,used the Real-Time PCR to assay expression of ?7n ACh R in liver tissue.Experiment 2 the protective effect of GTS-21 on acute liver injury in mice induced by LPS(1) 40 male C57BL/ 6 mice were randomly divided into normal control group(n = 10); blank control group(GTS-21 3mg/ kg, n = 10);model group(LPS 30mg/kg, n = 10); GTS-21 group(LPS 30mg/kg +GTS-21 3mg/kg, n = 10), adaptation feeding one week.the normal control group and model group treated as above, the model group was intraperitonealld GTS-21 3mg/kg, do not inject LPS, GTS-21 group was intraperitonealld GTS-21 3mg/kg before modeling(LPS 30mg/kg)30min.crificed all the mice after modeling 6 hours,(no mice died in each group), Each group selected 8 samples randomly,then observed the gross morphology of the liver, serum ALT, AST levels and liver tissue IL-18,IL-1?, HMGB1 content and ?7n ACh R expression levels as the xperiment1.(2) 30 male C57 BL / 6 mice were randomly divided into model group(n = 15), intervention group(n = 15), adaptive feeding one week, the model builded and intervention were treated as above, then observe the symptoms and the survival situation of mice in each group within48 h.counted the number of surviving mice at each time point, draw survival curves and do analysis about survival rate of each group.Results Experiment 1 study on the pathogenesis of ?7n ACh R in acute liver injury by LPS(1)Compared with normal control group, liver pathology of model group all have Different degree of pathological changes,the most significant lesions at 6 h lesions.compared with model group, thenormal control group liver pathology index was 0, the model group 6h was0.40,compared with other model subgroups,the difference was statistically significant(p <0.05).(2) compared with normal control group, each model subgroups serum AST,ALT levels were increased, and the differences were statistically significant(P <0.05), the model group six hours AST(378.37 ± 59.27 vs 170.00 ± 55.88, p = 0.011), ALT(132.58 ±45.53 vs 56.93 ±10.92, p = 0.002) increased most significantly;compared with other model subgroups,the difference was statistically significant(p<0.05).(3) compared with normal control group, the liver tissue IL-18,HMGB1, IL-1? levels were increased in each model subgroup, the differences were statistical significance(P <0.05), in which IL-18(393.83±60.18 vs 86.87±10.12),IL-1?(3988.92±253.25 vs 1148.15±66.00)after modeling 6h increased most significantly, compared with other subgroups the differences were statistically significant(P <0.05), while HMGB1(21.30±2.11vs4.99±1.07)12h after modeling increased most significant,compared to other subgroups, the difference was statistically significant(P<0.05).(4)Compared with normal control group,expression of ?7n AChR in each model subgroup decreased gradually, the differences were statistically significant(P <0.05).Experiment 2 the protective effect of GTS-21 on acute liver injury in mice induced by LPS(1) Compared with normal control group,liver pathology of model group seen a large number of inflammatory cell and necrosis,pathological changes significantly,compared with model group,the intervention group liver pathological changes improved significantly,normal control group liver pathology index was 0, intervention group compared with model group was(0.21 ± 0.16 vs 0.40 ± 0.00), the difference was statistically significant(p <0.05).(2) Compared with normal control group, model group AST(378.37 ± 59.27 vs 170.00 ±55.88 p = 0.011), ALT(132.58 ± 45.53 vs 56.93 ± 10.92 p = 0.002) was significantly increased, the difference was statistically significant(p<0.05); compared with model group, intervention group serum AST(222.51 ± 72.17 VS 378.37 ± 59.27 P = 0.011), ALT(61.12 ± 19.25 VS132.58 ± 45.53 P = 0.011) was significantly decreased, the difference was statistically significant(P = 0.011).(3) Compared with normal control group, model group liver tissue IL-18(393.83 ± 60.18 vs 86.87 ± 10.12),HMGB1(13.22 ± 1.88 vs 4.99 ± 1.07), IL-1?( 3988.92 ± 253.25 vs1148.15 ± 66.00) were increased, the differences were statistically significant(P <0.05), compared with model group, the intervention group liver tissue IL-18(132.05 ± 22.23 VS 393.83 ± 60.18 P = 0.02), IL-1?(1625.08 ± 120.38 VS 3988.92 ± 253.25 P = 0.011), HMGB1(7.73 ±1.75 VS 21.30 ± 2.11 P = 0.022) levels were significantly decreased, the difference was statistically significant(P <0.05).(4) Compared with normal control group,?7n ACh R expression of model group was decreased gradually, the difference was statistically significant(P <0.05),the intervention group liver tissue ?7n ACh R expression was significantly higher than the model group(2.11 ± 0.26 vs 1.11 ± 0.11), the difference was statistically significant(P <0.01).(5) survival rate of model mice was53.3%, the intervention group was 93.3%, the difference was statistically significant(P = 0.01).Conclusion(1) ?7n ACh R play an important role on the pathogenesis of acute liver injury in mice with LPS.(2) GTS-21 has a protective effect on acute liver injury in mice with LPS. |
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