| Objective: In this study,different concentrations of Xiaozhiling injection were used to construct a rat model of surface fistula,and the feasibility of modeling was explored by observing different indicators.Methods: Twenty-five healthy and clean SD rats were randomly divided into 5 groups,5 rats in each group(A spring gauze was injected into the neck and back of rats in groups A,B,C,D and E with 2%,3% and 4% Xiaozhiling injection,1 * 109 cfu/m E.coli bacterial solution and 0.9% normal saline,and the injection volume of each rat was 1 ml).Surface fistula modeling was performed,and on the 30 th day after modeling,spring gauze was removed from the neck and back of rats and continued to be observed until the 40 th day after modeling.Observe and record the general condition,body temperature,and local wound conditions of rats after modeling,MRI examination results of fistula formation in rats,pathological changes and protein expression of IL-6,IL-1β and TNF-α in the tissues of the modeling site of rats.Results:(1)General observation: From the 1st day to the 30 th day after model established,the rats in group A were in good spirits,with slightly scattered hair with white color,decreased gloss and there were no significant changes in activity,eating,water intake and stool and urine.The rats in group B were slightly poor in spirit,scattered in hair,slightly yellow in color,decreased gloss,decreased in activity,eating and drinking and no significant changes in stool and urine.The rats in group C were in poor spirits,with scattered hair,yellow and lusterless color,decreased activity,food and water consumption,and increased stool and urination frequency.The rats in group D were depressed,their hair was disordered,yellow and lustless,their activities and diet were significantly reduced,their aggressiveness was strong,and the number of stools and stools was significantly increased.The rats in group E were in good spirits,their hair was slightly scattered,white and glossy,and the amount of activity,food consumption,water consumption,and stool and urine did not change significantly.On the 30 th day after model established,the spring gauze was removed and observed until the 40 th day after model established,the mental status of rats in each group gradually returned to normal,the hair recovered luster,the activity level and dietary intake were increased than before,and the urine and defecation conditions returned to normal.(2)Temperature record: 1 hour,1st and 3rd day after modeling,the temperature of rats in group D was significantly higher than that in the other four groups,and the difference was statistically significant(p<0.05),On the 7th,14 th,21st,30 th and 40 th day after model establishment,there was no significant difference in body temperature between group D and groups A,B,C and E(p<0.05).There were no differences in body temperature among rats in groups A,B,C,and E throughout the modeling process(p<0.05).(3)Local wound condition record:1st day after model establishment,except that no obvious secretion was observed in the local wound of rats in group E,a small amount of secretion exudation was observed in the local wound of rats in the other groups,of which a small amount of secretion was observed in the local wound of rats in groups A,B and C,which was pale and thin,the local wound secretion of rats in group D was small,white and slightly sticky.The 3rd day after molding,the local wound secretion of rats in group A was small,white and thin,while the local wound secretion of rats in group B was slightly increased,white and thin.The local wound secretion of rats in group C was more,white and thick,while the local wound secretion of rats in group D was significantly increased,yellow and white and thick,while the local wound secretion of rats in group E was small,white and thin.The 7th day after molding,the local wound secretion of rats in group A decreased compared with that before,and the surrounding granulation tissue proliferated and showed light red.The local wound secretion of rats in group B had a small amount of secretion,which was white and slightly sticky,and the surrounding granulation tissue proliferated and showed light red.The local wound secretion of rats in group C was more,which was yellow and white and sticky,and the surrounding granulation tissue proliferated and showed light red.The local wound secretion of rats in group D was more,which was yellow and sticky,and the surrounding granulation tissue proliferated and showed red.There is no obvious secretion and granulation tissue on the local wound of rats in group E,which is light red.The 14 th day after molding,no secretion was found on the local wounds of rats in groups A,B and E,and granulation tissue hyperplasia was light red.In group C,there was a small amount of secretion on the local wound of rats,which was white and thin,and granulation tissue proliferated around it,showing bright red.In group D,there were many secretions on the local wounds of rats,which were yellow and white,thin in texture,and the surrounding granulation tissue proliferated,bright red in color.No secretion was observed in the local wound of rats in groups A,B and E,and the granulation tissue proliferated and was reddish;there was a small amount of secretion in the local wound of rats in group C,which was white in color and thin in quality,and the granulation tissue proliferated around it and presented bright red.There were more secretions in the local wound of rats in group D,which were yellowish white in color and thin in quality,and the granulation tissue proliferated around it and bright red in color.On the 21 st day after model establishment,the local wound surface of rats A and B was dry,and granulation tissue with high protrusion was observed around it;no obvious secretion was observed on the local wound surface of rats in group C,and granulation tissue with high protrusion around it.A small amount of secretion was observed on the local wound surface of rats in group D,which was transparent in color,thin in quality,and granulation tissue with high protrusion around it.The wound surface at both ends of the local wound surface of rats in group E was dry,and granulation was flat.On the 30 th day after model establishment,after removing the spring gauze of all rats,the local wound surface of rats in groups A,B,C,D and E was dry,and a hard cord was palpable in the subcutaneous deep part of the wound at both ends.On the 40 th day after model establishment,a hard cord was still palpable in g roups C and D,and no hard cord was palpable in groups A,B,and E.(4)MRI examination results: All rats in groups C and D showed imaging manifestations of fistula.(5)Pathological examination results: The morphology of muscle fibers in groups A,B,and E is relatively regular.Aggregated muscle cells can be seen in the muscle bundles of groups A and E,and no obvious inflammatory cell infiltration is observed in the stroma.Loose arrangement of muscle cells can be seen in the local muscle bundles of group B,and scattered inflammatory cell infiltration can be seen in the stroma;No obvious muscle cells were found in the muscle bundles of groups C and D,with a large amount of proliferative granulation tissue,fibroblasts,newly formed capillaries,and a small amount of multinucleated cells visible.Among them,group C showed more inflammatory cell infiltration,while group D showed a large number of inflammatory cell infiltration.(6)WB results: Compared with group E,the expression of IL-6,IL-1β and TNF-α protein in the tissues of the modeling site of rats in groups A,B,C and D was increased,and the difference was statistically significant(p<0.05),with group D being the highest,followed by groups C,B and A(p<0.05).Conclusion: 1.Xiaozhiling injection can successfully construct the rat model of surface fistula,of which 4% Xiaozhiling injection is the effective concentration.2.By comparing the general condition,local wound condition,analysis of pathological examination results and expression of inflammatory factor proteins in the tissue of the modeling site in the E.coli bacterial solution,4% Xiaozhiling injection can construct the surface fistula model of the rats close to the liquid phase of E.coli bacterial solution.3.Compared with the limitations of E.coli,such as low biosafety,difficult preparation,high cost and price,Xiaozhiling injection has the advantages of convenient access and high safety,so it is recommended to use 4% Xiaozhiling injection to construct the rat surface fistula model. |
PDF Full Text Request