| Objective:to observe the effects of Huaxian capsule on the expression of T cell CD69 and CD4+CD25+Treg in rats with sepsis,and to provide theoretical basis and relevant laboratory indexes for the treatment of sepsis immune dysfunction by huaxian capsule.Methods:135 healthy SD male rats were randomly divided into two parts:Pre-experimental part:15 rats in the half cecal ligation group and 15 rats in sham operation group.Experimental part:15 rats in sham operation group;model group(24h group.48h group.72h group 15 rats in each group);Huaxian capsule treatment group(24h group.48h group.72h group 15 rats in each group).(1)the steps of modeling:after 24 hours of preoperative fasting(except water)of rats in each group,10%chloral hydrate(0.4ml/100 g)was injected into the abdominal cavity.After the effect of anesthesia,rats keeping the supine position were fixed on the operating table,and routine disinfection after abdominal preparation.Cut the abdominal skin about 2cm along the midline of the abdomen;? the half cecal ligation:after finding the cecum,ligature of the half cecum at a distance of 1-2cm from the blind side of the cecum.At the same time,14 needle was used to puncture the cecum for 2 to 3 times.After the small amount of feces was extruded,the cecum was returned to the abdominal cavity to close the abdomen.? Sham operation group:the distal cecum and mesentery were separated,and no ligation was performed.The part of experimental research:Both the model and the treatment group were treated with half cecal ligation.(2)the processing factors:the treatment group was given huaxian capsule of 200mg/kg by gavaging(the first time was 2h after the operation,and the second time was 12h after the operation,once every 12h).Sham group and model group:same volume physiological saline(the first time was 2h after the operation,and the second time was 12h after the operation,once every 12h).(3)Collection and detection of the experimental specimens:?pre-experiment part:observe the general state of the rats in the half cacal ligation group and the sham operation group,collect the tail blood for blood culture in 24h and dissect the rats in each group;?experiment part:the sham operation group carried out live killing of rats in 24h and collected blood samples for testing;and rats in model group and treatment group(24h,36h,72h)were used to collect blood.The T cell CD69,CD4+CD25+Treg in peripheral blood were detected by flow cytometry.Result:1.Pre-experimental part:the results of 24h blood bacteria culture in the half cecal ligation group of the cecum were positive,and after dissection,the accumulation fluid was visible in the abdominal cavity,and the necrosis of the cecum was black.The results of blood culture in sham operation group were negative,there was no effusion in the abdominal cavity and no necrosis in the intestinal canal.There were no dead rats in the sham operation group and the half cecal ligation group in 24h.2.Experimental part:T cell CD69:at the time point of 24h,48h and 72h,the differences between the model group and the treatment group and the sham operation group was statistically significant(P<0.01)The differences of the model group at 24h,48h and 72h were statistically significant(P<0.01).The differences in the treatment group of 24h,48h and 72h were statistically significant but not remarkable(P<0.05).Comparison between groups:the difference between the model group and the treatment group in 72h was statistically significant(P<0.05).CD4+CD25+Treg:at the time point of 24h,48h and 72h,the differences between model group and treatment group and sham operation group was statistically significant(P<0.01).The differences of the model group at 24h,48h and 72h were statistically significant(P<0.01).The differences in treatment of 24h,48h and 72h were statistically significant but not remarkable(P<0.05).Comparison between groups:the difference between the model group and the treatment group in 72h was statistically significant(P<0.05).Conclusion:1.Pre-experimental part:it proved that the rat model of sepsis was successfully induced by half cecal ligation.The sham operation group showed that the separation of distal cecum and mesentery did not lead to sepsis,and the half cecal ligation was the key to induce sepsis model.2.Experimental part:T cell CD69:at 72h,the difference between the treatment group and the model group was statistically significant(P<0.05),at 72h huaxian capsule was able to improve the expression of CD69 in rats with sepsis.CD4+CD25+Treg:at 72h,the difference between the treatment group and the model group was statistically significant(P<0.05),at 72h huaxian capsule could reduce the expression of CD4+CD25+ Treg in sepsis rats. |
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