Study On Expression Of Aquaporins In Rotavirus Enteritis Treatment By Drug

Objective1. The building of rotavirus enteritise model2. To study the mechanism of rotavirus enteritis whether correlated with the alteration of aquaporin1,3,4,8mRNA.3. To study the alteration of AQP1,3,4,8mRNA in jejunum,ileum and colon of rotavirus enteritis infant mice which treated by drug.Materials and Method1. SubjectsInfant Kunming mice, ,supplied by Zhongshan university animal house.2. RotavirusHuman rotavirus supplied by Guangzhou children hospital where the patients diagnosed rotavirus infection in December 2007.Rhesus rotavirus supplied by China Medical College Institute of Biology.3. The building of rotavirus enteritise modelIn experimental condition,using 150ul TCID50 is 1×104 rotavirus to infect mice aged 6days,which is the diarrhea group(B group).Using 150ul not contain rotavirus to make mice driven,which is the blank group(A group).4. The building of drug treatment modelAfter giving rotavirus to mice 24h,48h,using drug(1.86g/ml) to treat the mice ,as the C group and D group.Three times a day, 50ul for every times.5. Intestinal biopsyTaking the enteral duct into three parts. Picking up the jejunum, ileum and the colon. The duct stored in liquid nitrogen.6. Fluorescent quantitative PCR to test the expression of aquaporin 1,3,4,8mRNA in enteral duct。4) Abstraction of RNA in enteral duct tissue.5) Reverse transcription of RNA.6) Fluorescent quantitative PCR to test the expression of aquaporin 1,3,4,8mRNA.7. Statistical analysis data were analyzed by SPSS version 11.0.Results1. The building of rotavirus enteritise modelApparent diarrhoea was observed in mice after infecting SA11 at 2 days,RV antigen could be detected in stool samples collected from SA11 infected mice at least from the second to sixth day post-infection.But mice infected by human rotavirus had not been diarrhoea, and RV antigen could not be detected in stool samples.2. The clinical manifestation of rotavirus enteritis model after treatment by drugThe diarrhea group (C group),one third infant mice diarrhea after infection rotavirus 48 hour.The main clinical manifestation is waterly stool.The rate of diarrhea mice is 33%.After infection rotavirus 3rd ,4th ,diarrhea was moderated .After infection rotavirus 5th to 7th ,all the mice were no diarrhea.The diarrhea group(D group),all the mice diarrhea after infection rotavirus 48 hour.The clinical manifestation is waterly stool.The rate of diarrhea mice is 100%.After infection rotavirus 3rd,4th,5th,diarrhea was moderated.After infection rotavirus 6th ,7th,all the mice were no diarrhea.3. The results of Fluorescent quantitative PCR . 1) infant mice's jejunum ileum and colon express the aquaporin 1,3,4,8mRNA.2) the diarrhoea group's expression of AQP8mRNA in jejunum is higher than the blank group's(t=2.304,P<0.05),3) the expression of AQP1mRNA in ileum is lower than the blank group's(t=-2.25,P<0.05).4) In jejunum and ileum ,the other AQPs are no difference in diarrhea group and blank group.5) In colon ,the expression of AQP1,3,4,8mRNA has no correlation with diarrhea group and blank group(P>0.05)6) In jejunum ,the expression of AQP1,3,4,8mRNA are no difference in drug group(C and D group) and blank group(P>0.05).7) In ileum , the expression of AQP1,3,4,8mRNA are no difference in drug group(C and D group) and blank group(P>0.05).8) In colon , the expression of AQP1mRNA in C and D group was higher than the blank group(t=-3.470,P<0.05;t=-3.320,P<0.05). The expression of AQP3,4,8mRNA are no difference in drug group(C and D group) and blank group(P>0.05).Conclusion1. Infant Kunming mouse is sensitive to SA11, but not sensitive to human rotavirus.2. The higher expression of AQP8mRNA in jejunum of diarrhea infant mice,indicates that AQP8 joins the absorption and secretion in intestine.The expression of AQP8 increasing compensativly ,can decrease the degree of diarrhea in infant mice.3. The lower expression of AQP1mRNA in ileum of diarrhea infant mice,indicates that the lower AQP1 expression in ileum,the smaller water absorpt.Ileum absorption function is disordered.As a result,absorption of water is reduced,thus loose stool and diarrhea occur. 4. The level of AQP1mRNA changement is stronger than AQP8mRNA's. The major clinical situation of diarrhea group of mice is watery stool.5. There is no relationship between rotavirus enteritis and aquaporin 1,3,4,8 in colon.6. The higher expression of AQP1mRNA in colon of the drug groug(C and D group) , indicate that the drug can increase the expression of AQP1mRNA in colon.Infant mice make water absorption increase in colon through raising the expression of AQP1,in order to decrease the waterly stool .7. In the study,there is no alteration of AQP3,4,8mRNA in intestinal tract of rotavirus enteritis infant mice which treated by drug .8. Rotavirus enteritis mice treated by drug in different time,which effected the pathogenesis of the enteritis.After 24h infecting rotavirus,mice were treated by drug.It could decrease the rate of diarrhea (C group:the rate was 33%;D group:the rate was 100%).At the same time,it could shorten the diarrhea pathogenesis(C group:the pathogenesis was 0—3days;D group:the pathogenesis was 2—4days).