| Objective As an actin-binding protein,gelsolin(GSN)is involved in the life processes of various cells and is closely related to pathophysiological processes such as trauma,sepsis,immunity and inflammation.Decreased levels are associated with increased mortality and increased mortality in patients with severe burns and sepsis.Studying its changing laws and pathophysiological effects will help to deepen the understanding of the pathogenesis of sepsis and provide potential intervention targets for the regulation of immune response.In this study,GSN was used to treat severely burned mice in vivo,to observe its changes and influence on T cell immune function and to explore its mechanism,aiming at providing new ideas of prevention and treatment of sepsis and multiple organ dysfunction syndrome.Methods Experiment 1:80 male BALB/c mice were randomly divided into a false injury group and a bum group,with 40 rats in each group,n=8.The burn group mice caused 15%TBS A ? degree bum on the back(hereinafter referred to as burn).Immediately after the injury,1 ml of normal saline was injected subcutaneously,and the back was coated with appropriate amount of iodophor to prevent infection once a day.The mice in the sham-injured group were injured,and no rehydration or extra iodine was applied after the injury.Eight mice in each group were taken at 0(immediately),8,24,48,and 72 h after injury,and the spleens were aseptically removed.The proliferation activity of spleen T 1ymphocytes was detected by thiazolyl blue colorimetric assay.The spleen tissues were detected by ELISA.The content of GSN mRNA in spleen was detected by real-time quantitative RT-PCR.Experiment 2:150 male BALB/c mice were randomly divided into normal control group(A),false injury group(B),bum group(C),low dose group(D),and high.Dosing group(E),30 per group,n=6.GSN 0.05 mg(0.1 ml)/only,0.5 mg(0.1 ml)/only,were administered immediately after the injury in the low-dose group and the high-dose group.At the 0(immediate),8,24,48,and 72 h post-injury,6 mice in each group were taken,serum,spleen and liver were aseptically collected,and GSN content in serum,spleen and liver tissues was detected by ELISA.Fluorescence quantitative RT-PCR was used to detect the expression of GSN mRNA in spleen and liver tissues.Mortality test animals group:120 male BALB/c mice were randomly divided into a false injury group(B),a bum group(C),a low dose group(D),and a high dose group.(E),30 rats in each group,group C,group D,group E were injected subcutaneously with 1 ml of saline solution 8 h after injury,and the death time of the animals was observed and recorded until 168 h after injury.Experiment 3:150 male BALB/c mice were randomly divided into normal control group(A),false injury group(B),burn group(C),low dose group(D),and high.Dosing group(E),30 per group,n=6.At the 0(immediate),8,24,48,and 72 h post-injury,6 mice in each group were taken,and the spleen was aseptically removed.The proliferation activity of spleen T 1ymphocytes was detected by thiazolyl blue colorimetric assay.IL-2,IL-4 and IFN-? in spleen were detected by ELISA.Results Experiment 1:The proliferative activity of spleen T 1ymphocytes,the content of GSN in spleen tissues and the expression of GSN mRNA in spleen tissues of the spleen mice were significantly higher than those in the burn group(P values were less than 0.05)at 8,24,48 and 72 hours after injury.The proliferative activity of spleen T 1ymphocytes in burn group was 0.117±0.036 8 h after injury,which was significantly lower than 0.728±0.067,0.563±0.069,0.506±0.009 and 0.593±0.069 at 0,24,48 and 72 h after injury.All were less than 0.05);the GSN content(11.25±5.281)pg/mg in the spleen tissue of the burn group was significantly lower than that of the group at 0,24,48,72 h(37.74±2.858),(19.98±6.233).),(24.68±7.336),(24.59±5.077)pg/mg(P value is less than 0.05);the expression of GSN mRNA in spleen tissue of burn group was 0.807±0.064 8 h after injury,which was significantly lower than that of group internal injury.0.936 ± 0.123,0.625±0.091,0.744± 0.104,and 0.821± 0.072 at 24,48,and 72 h(P values are less than 0.05).The GSN content and GSN mRNA expression in the spleen of the burn group were significantly positively correlated with the proliferation of T 1ymphocytes at 0,8,24,48,72 h after injury(P value was less than 0.05).Experiment 2:Serum GSN content,spleen GSN content and mRNA expression,liver tissue GSN content and mRNA expression were significantly higher in the injured group than in the burn group,low dose group and high dose group at 8,24,48 and 72 hours after injury.(P values are less than 0.05).The levels of serum GSN,GSN content and mRNA expression,liver tissue GSN content and mRNA expression in the bum group were significantly lower than those in the high dose group at 8,24,48 and 72 hours after injury(P value was less than 0.05).The serum GSN content(851±32)pg/mg and the serum GSN content(956±87)pg/mg in the high-dose group were significantly higher than those in the low-dose group(662±67),(803±197)pg/mg(P values are all less than 0.05).The levels of GSN and mRNA in spleen and liver tissues of mice in high-dose group were significantly higher than those in low-dose group(P<0.05).Severe bums caused mice to die within 72 hours after injury.The mortality rate of the bum group and the low dose group were 70%,which were significantly higher than those of the high dose group(43.33%)(P values were less than 0.05),the survival time(19.86±15.47)h in the burn group was significantly lower than that in the low dose group(30.76±18.46)h(P<0.05).Experiment 3:The proliferation activity of spleen T lymphocytes was significantly higher in the spleen mice than in the burn group,the low dose group and the high dose group at 8,24,48 and 72 hours after injury(P value was less than 0.05).The proliferative activity of spleen T 1ymphocytes in the bum group was not significantly different at 8,24,48,72 h after injury(P value was greater than 0.05).The spleen T 1ymphocyte proliferation activity in the bum group was significant at 8 and 24 h after injury.Lower than the high dose group(P values are less than 0.05).The levels of IL-2 secreted by spleen T 1ymphocytes in the burn group were lower than those in the sham,low-dose and high-dose groups(P values were less than 0.05),and the spleen T of the injured group was 8 h after injury.IL-2 secretion from 1ymphocytes was higher than that in low dose group and high dose group(P value was less than 0.05).In the high dose group,spleen T 1ymphocytes secreted IL-2 and low at 0,8,24,48 and 72 after injury.There were no significant differences in the dose group(P values were greater than 0.05).The secretion of IL-4 from spleen T 1ymphocytes in the burn group was significantly higher than that in the sham injury group(P value less than 0.05),IFN-? secretion and ratio(IFN-?/IL-4)at 8,24,48,72 h after injury.Significantly lower than the false injury group(P value is less than 0.05).There were no difference significant in the content of IL-4 between the spleen T 1ymphocytes and the bum group at 0,8,24,48,and 72 h after injury in the low-dose group(P value was greater than 0.05),and spleen T 1ymphocyte secretion was observed at 8,24,and 48 h after injury.IFN-? was significantly higher than burn group(P value was less than 0.05),and the 8h post-injury ratio(IFN-?/IL-4)was significantly higher than that of bum group(p<0.05).The secretion of IL-4 from spleen T 1ymphocytes in the high-dose group was significantly lower than that in the bum group and the low-dose group(P value was less than 0.05).The spleen T 1ymphocytes secreted IFN-?at 8,24,48 h after injury.The amount and ratio were significantly higher than those in the bum group(P values were less than 0.05)Conclusions Experiment 1:Severe bums resulted in the proliferation of spleen T 1ymphocytes,GSN content and gene expression in mice,and GSN content and GSN mRNA expression were positively correlated with T 1ymphocyte proliferation activity,and all three were 8 h after injury.To the lowest,the best time point for GSN intervention after severe bums can be selected 8 hours after injury.Experiment 2:GSN was consumed after severe bums.GSN intervention can significantly reduce the mortality and prolong the survival time of severely burned mice.The intervention effect of GSN is related to the dose.High dose GSN can significantly inhibit the serum GSN content caused by severe burns The spleen and liver GSN content and GSN mRNA expression decreased,and the intervention effect was significantly better than the low dose group.Experiment 3:GSN can significantly reduce the imnunosuppressive effect caused by severe burns,and can promote the shift of T 1ymphocytes to Thl after severe burns.This drift direction will indicate that the immune status of the body shifts from inhibition to hyperactivity. |
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