The Study Of The Effect Of Filling The Immobilized Urease In Colostomy Pouch On The BUN In Rats

Objective To established a model of acute renal failure to evaluate the effect of blood urea nitrogen(BUN)on filling the immobilized ureas in colostomy pouch.Methods Used gelatin as carrier for enzyme immobilization and immobilized urease in glutaraldehyde cross-linked gelatin membrane.All the experimental rats were divided into six groups:normal control group(group A,total 3),sham-operated group(group B,total 7),colostomy pouch operated group(group C,total 7),renal failure group(group D,total 7),renal failure-colostomy pouch operated group(group E,total8),renal failure-colostomy pouch operated-fill immobilized urease group(group F,total 8).Preoperative enema for two days.After intraperitoneal injection of anesthesia with nembutal,the sham-operated group(group B)suture after enterocoelia is opened and don’t have any special operation.Open the rats’ enterocoelia in the colostomy pouch operated group(group C)and located a length of abundant blood supply colon about 1.5cm.To protect this length of colon and cut through both ends of it.The colon-colon anastomosis was interrupted sutured outsaid mucous membrane layer.Use the same way to operate on renal failure-colostomy pouch operated group(group E).Open the rats’ enterocoelia in the renal failure-colostomy pouch operated-fill immobilized urease group(group F).To suture one end and put the immobilized urease from the other side,then sutured the end.The colon-colon anastomosis was interrupted sutured outsaid mucous membrane layer.All of the surgical rats were injected 100 thousand UI of penicillin in muscle to prevent the infection.Group A and D were injected the saline in same dose.Fasted all the surgical rats and fed them with 10%glucose in the three days after operation.After the third day,the rats was fed with liquid diet.Disinfected the surgical rats twice a day on time and last a week after operation.On the fifth day,keep the rats off water for 12 hours.10mL/kg 50%glycerol were injected in the group D,E and F on both posterior limbs.The same dose saline were given to the group A,B and C.Collected 1ml blood from endocanthion on 24 h,48 h and 72 h after injected glycerol.Tested the levels of BUN.All the experimental rats were killed by cervical dislocation,and collected the immobilized urease that filled in colon.Detected the content of ammonia with titration.The kidney and colon were rapidly extracted,then preserved in paraformaldehyde.Observed pathological changes of kidney tissue and colon with HE staining.Results The content of BUN in the group D is significantly higher than that in the group A、B and C in 24 h(P<0.05);The BUN arrived the highest content on 72 h.The content of BUN in group D is significantly higher than group A、B and C after 48 h(P48h=0.004,P72h=0.007).It showed that the BUN begun to increase after injection of glycerol and kept this trend in 72 h.The BUN of renal failure-colostomy pouch surgery showed the trend of rising as simple renal failure group and it is observably higher than that in the groupA、B and C in 72 h(P24h=0.002,P48h=0.003,P72h=0.003).The BUN in the group F is higher than that in the group A、B and C in 24 h,but there is no significant difference.In the other hand,the content of BUN is significantly lower than the group E(P24h=0.006).The BUN is significantly lower than that in the group A、B and C in 24～48 h(P48h<0.05).Compared with the group E,the BUN is significantly lower(P48h=0.000).The BUN rose in 48～72 h,and near to the group A but there is significant difference(P72h=0.03).In the same time the content of BUN is significantly lower than the group E(P72h=0.01).It showed that the BUN in group F had been adsorbed in 24 h and arrived the lowest point in 24～48 h.The ammonia of the immobilized ureas in colostomy pouch is in different degrees and there is no obvious stimulation of colostomy pouch.Conclusion 1.The immobilized urease has good adsorption for the BUN of renal failure rats and it directly adsorbed the urea nitrogen.2.There is no obvious stimulation of colon after holding the immobilized urease in colostomy pouch during 72 h.3.The immobilized urease in the colon of rats can stabily and directly adsorbed the urea nitrogen.