The Study Of Daochichengqi Decoction In The Treatment Of Escherichia Coli Induced Rat Acute Pyelonephritis (APN)

Objective: in this paper,we make an APN rat model by the method of direct bladder injection of Escherichia coli without ligation ureter,in order to observe the changes of the Renal tissue,temperature,urine volume,kidney weight coefficient,blood routine,urine routine,urine bacteria culture,interleukin-2(IL-2),C reactive protein(CRP),beta N-acetyl Glucosaminidase(NAG)and secretory immunoglobulin A(sIgA)of rats.Finally,find out the effect of Daochichengqi Decoction on APN and its action mechanism,and then providing the experimental basis for clinical application and teaching of traditional Chinese medicine.Acute pyelonephritis(APN)from a variety of pathogenic microorganisms infections directly cause renal tubular,interstitial renal inflammation.Common clinical urinary frequency,urgency,dysuria,abdominal pain,back pain,rib ridge angle tenderness,percussion pain,accompanied by chills,fever,nausea,vomitin g,elevated white blood cell count and other symptoms of systemic infection.The basic pathology is characterized by renal pelvis mucosa congestion and edema,purulent discharge surface,inflammatory lesions in renal cortex,renal tubular epithelial swelling,loss necrosis,leucocyte infiltration,inflammation healing scar formation,but renal ball generally less impaired [1].In ancient medical books,although there is no clear on APN records,but according to the body of the clinical manifestations of hot,abdominal fullness,constipation,dry mouth,cleft lip,with poor urine trickle down,drowning when pain,rine color red,polydipsia,red tongue,rapid pulse and so on,they can be attributable to the category of “linzheng” “longbi” “xulaoyaotong” and so on [2].In view of the clinical manifestations of APN,traditional Chinese medicine starts from the theory of “the exterior-interior relationship between the kidney and the bladder” which reflects the integration concept of viscera related,the pathogenesis characteristics can be summarized as the kidney deficiency bladder heat[3].The study according to the traditional Chinese medicine pathogenesis characteristics of APN,the theory of “the exterior-interior relationship between the kidney and the bladder” and the modern pharmacological research of traditional Chinese medicine,choosing Daochichengqi Decoction for the treatment of APN rats.Daochichengqi Decoction from Wenbing Tiaobian of Wu Jutong,Article 17 of Wenbing Tiaobian-zhongjiao article: "and left vein firmly adhere,urine pain,when vexed thirsty very,It can be treated with Daochichengqi Decoction".The parties attending Yangming solid organs and intestinal Heat Syndrome,diarrhea stool that is able to pass,and Tom Lee urine,evil two solutions colon and bladder,as "gut Tongzhi two" of the law[4].Therefore,the disease can be treated with Daochichengqi Decoction which can gankuhuayin Lee urine and capture legitimate and piay a role of clearing heat and purging fire,dampness Tonglin[5].Through observing the changes of the renal tissue,temperature,the amount of urine,kidney weight coefficient,blood routine,urine routine,urine bacteria culture,interleukin-2(IL-2),C reactive protein(CRP),beta N-acetyl Glucosaminidase(NAG)and secretory immunoglobulin A(sIgA)of rats,we determine the mechanism of the treatment of APN with Daofchichengqi Decoction to provide experimental basis for the clinical treatment of APN and teaching of traditional Chinese medicine.Methods:1 The experimental group: Select 28 male SD rats of healthy and clean grade at weight of 250±20g,feeded adaptively 3 days,free access to water during the experiment,eating normal diet.They were divided into 4 groups with the principle of random allocation: normal group(Normal control group,K=n6),model group(Model group,M=n6),Daochichengqi Decoction group(Daochichengqi Decoction group,D=n8),levofloxacin(levofloxacin Hydrochloride group,L=n8).Except the normal group,other groups were used the method of the direct bladder injection of Escherichia coli(0.6mL)without ligation ureter to make an APN model of rat.2 Experimental drug delivery: Four groups were modeling in continuous administration for 5 days,dosage was calculated according to human and rat drug conversion ratio coefficient.D group was given Daochichengqi Decoction(4.9g/kg/d),L group was given levofloxacin(0.06g/kg/d),N group and M group were given the same amount of normal saline(8mL/kg/d).They were administered orally once a day.3 Detection of indicators: at nine clock everyday morning,rats were measured rectal temperature by electronic thermometer.And the rats were placed into metabolic cages to collect 24 hours urine at the fourth day,observe the urine changes of different groups.When the rats were killed,we measured the rat body weight and kidney weight,and then calculate the kidney weight coefficient.Blood from the abdominal artery,and then detect the changes of blood routine,interleukin-2(IL-2),C reactive protein(CRP).Urine was extracted from the bladder under sterile conditions,testing urine bacteria culture,urine routine,beta N-acetyl Glucosaminidase(NAG)and secretory immunoglobulin A(sIgA).Put the most kidney tissue that cut longitudinaly from the left side into 10% poly formaldehyde,dehydrated,embedded in paraffin,HE staining,then observe the structures and changes of renal under light microscope.Results:1 The results of indicators of rats in each group The results of temperature,compared with the normal control group,the level of temperature in model group,Daochichengqi Decoction group and levofloxacin group rats after modeling rise,the temperature of model group was increased significantly,but no significant difference(P>0.05).Compared with the model group,the level of temperature in Daochichengqi Decoction group and levofloxacin group rats decreased,the level of temperature in levofloxacin group decreased significantly,but there was no significant difference between Daochichengqi Decoction group and model group(P>0.05).The results of 24 h urinary volume,compared with the normal control group,the level of 24 h urinary volume in model group,Daochichengqi Decoction group and levofloxacin group rats increased,24 h urinary volume of model group was increased significantly,but no significant difference P>0.05).Compared with the model group,the level of 24 h urinary volume in Daochichengqi Decoction group and levofloxacin group rats decreased,but no significant difference(P>0.05).There are no obvious differences between Daochichengqi Decoction group and levofloxacin group(P>0.05).The results of kidney weight coefficient,compared with the normal control group,the level of Left kidney weight coefficient in the model group rats increased(P<0.05).Compared with the model group,the level of Left/Right kidney weight coefficient in Daochichengqi Decoction group rats decreased(P<0.05),the level of Left/Right kidney weight coefficient in levofloxacin group rats decreased significantly(P<0.01).There are no obvious differences between Daochichengqi Decoction group and levofloxacin group(P>0.05).The results of blood routine,The results of WBC and N,compared with the normal control group,the level of WBC in the model group rats increased(P<0.05);Compared with the model group,the level of WBC and N in levofloxacin group rats decreased significantly(P<0.01),the level of WBC and N in Daochichengqi Decoction group rats decreased,but no significant difference(P>0.05).Compared with the Daochichengqi Decoction group,the level of WBC in the levofloxacin group rats decreased(P<0.05).The results of RBC and HB,compared with the normal control group,the level of RBC and HB in the model group rats decreased(P<0.05);But there was no significant differences between Daochichengqi Decoction group,model group and levofloxacin group(P>0.05).The results of PLT,There was no significant differences among the four groups(P>0.05).The results of urine routine,The results of U-LEU,compared with the normal control group,the level of U-LEU in the model group rats increased(P<0.05);Compared with the model group,the level of U-LEU in Daochichengqi Decoction group rats decreased(P<0.05),the level of U-LEU in levofloxacin group rats decreased,but no significant difference(P>0.05).There are no obvious differences between Daochichengqi Decoction group and levofloxacin group(P>0.05).The results of Urine occult blood,compared with the normal control group,the level of Urine occult blood in the model group rats increased significantly(P<0.01);But there was no significant differences between Daochichengqi Decoction group,model group and levofloxacin group(P> 0.05).The results of urine bacteria culture,compared with the normal control group,the positive rate of urine culture in model group was statistically significant(P<0.05).Compared with the model group,the positive rate of urine culture in Levofloxacin group was statistically significant(P<0.05),the positive rate of urine culture in Daochichengqi Decoction group was not statistically significant(P>0.05).But there was no significant differences between Daochi chengqi Decoction group and the levofloxacin group(P>0.05).The results of sIgA,compared with the normal control group,the level of sIgA in the model group rats decreased significantly(P<0.01);Compared with the model group,the level of sIgA in levofloxacin group and Daochichengqi Decoction group rats increased significantly(P<0.01);Compared with the Daochichengqi Decoction group,the level of sIgA in the levofloxacin group rats indecreased(P<0.05).The results of NAG,There was no significant differences among the four groups(P>0.05).The results of IL-2,compared with the normal control group,the level of IL-2 in the model group rats increased significantly,but no significant difference(P>0.05).Compared with the model group,the level of IL-2 in Daochichengqi Decoction group rats decreased significantly(P<0.05);But there was no significant differences between Daochichengqi Decoction group and levofloxacin group(P> 0.05).The results of CRP,compared with the normal control group,the level of CRP in the model group rats increased significantly(P<0.01).Compared with the model group,the level of CRP in Daochichengqi Decoction group rats decreased(P<0.05),the level of CRP in levofloxacin group rats decreased significantly(P<0.01).There are no obvious differences between Daochi chengqi Decoction group and levofloxacin group(P>0.05).2 Pathology morphological observation of renal tissue under the light microscopeThe structure of renal tissue of the normal control group rats is complete,there are no pathological changes;In the model group,the infiltration of lymphocytes and the small amount of eosinophils were seen in the renal pelvis and interstitial area,Interstitial area of capillary dilation,congestion,renal tubular necrosis;compared with the model group,the pathological changes of Daochichengqi Decoction group and levofloxacin group improve significantly,in the rat renal pelvic mucosa and interstitial area,the rare lymphocytes were infiltrated,the capillary dilation in the interstitial area was reduced,the renal tubular necrosis was relieved,and the blood vessel congestion was relieved.Conclusion:1 Daochichengqi Decoction could increase the level of secretory immunoglobulin A(sIgA),the level of White blood cell and neutrophil(WBC andN),interleukin-2(IL-2),C reactive protein(CRP),Urinary Leukocyte(U-LEU)will be reduced,and the renal pelvis submucosal,interstitial infiltration of neutrophils and lymphocytes,telangiectasia,congestion and edema also will be relieved,it has some protective effect on kidney,there are no obvious differences between Daochichengqi Decoction and levofloxacin(P>0.05)during the short-term therapeutic.It proves that Daochichengqi Decoction has certain therapeutic effect on APN.2 The mechanism of action of Daochichengqi Decoction on APN may relate to that it can qingre pee Tongfu to antibacterial anti-inflammatory and enhance the immune function of the local urethral.3 Experimental study confirms the Daochichengqi Decoction is through anti-inflammatory and enhanced local immune function of urethral play the joys of gankuhuayin Lee urine and capture legitimate,so as to achieve the purpose of clearing heat and removing dampness Tonglin.It provides experimental basis for teaching and clinical a pplication of traditional Chinese medicine.4 Experimental studies confirm the increase in urine sIgA levels and anti-inflammatory aspects,the efficacy of levofloxacin is superior to Daochichengqi Decoction,suggesting Clinical shall be based on the actual situation of the patient's condition,were selected for the Chinese and Western medicine treatment.But Daochichengqi Decoction therapeutic advantages in its entirety,as well as toxic side effects,less drug resistance or other adverse reactions and other advantages,clinical treatment APN can be used as adjunctive therapy to improve the clinical cure rate and reduce the recurrence rate.