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Expression of Aquaporin 3、4、8 in Colonic Mucosa of Rats with Slow Transit Constipation (STC)

Author: ZhiHui
Tutor: YuanWeiTang
School: Zhengzhou University
Course: Surgery
Keywords: aquaporin3 aquaporin4 aquaporin8 STC RT-PCR
CLC: R574.62
Type: Master's thesis
Year: 2011
Downloads: 88
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Abstract

Background and aimConstipation is a common clinical symptom,which prensents defecation reduces,difficult defecation,dry feces with little dejection,which influences the quality of life severely. As the aging population coming,constipation is attracting more attention as tumor,but it is still at the exploratory stage when dealing with constipation. The standard of RomenⅢwhich published in 2006, improved the dianosis of constipation,to promote the further studies on constipation. Constipation is induced by many factors including gastrointestinal disease,and other system disease in relation to gastrointestinal disease;many kinds of drug also induce constipation, Functional constipation (FC) is in the majority in constipation.On the whole,constipation includes Functional constipation (FC), Irritable bowel syndrome with constipation,(IBS-S),Functional delecation disorders. For the diagnosis of Functional disease,firstly organic disease and drug-induced disease must be excluded, Detailed clinical history,physical examination,and the examination of laboratory,iconography can provide important envidence for the diagnosis of constipation. After the organic disease is excluded,Functional constipation can be dibided into different styles according to the standard of RomenⅢand psychotherapy may be necessary for some anstipation patient together with mental symptom.After diagnosis of anstipation is made,FC can be divided into Slow transit constipation (STC), Outlet obstructive constip ation (OOC) and MIX according to the examination of intestinal motility and anorectal function. STC is a type of obstipation what feature are weaken of the gastrointestinal power and the lengthen of the time of slow transit of the content in gastrointestinal tract.The clinical symptoms are defecation reduces, little desire to defecation,difficult defecation and constipated,et al.Some researches on pathogenesis about STC reveal that STC is induced by many factors including the degeneration or reduction of enteric nerve,the degeneration of smooth muscle,the exception of gastrointestinal hormo ne unreasonable diet,the degeneration of cajal parcemaking cell and the factors of mental and hereditary et al.Among many researches,there is little investigation about Aquaporin and constipation.The water in the content of bowel is dried severely and the content becomes dry which makes it difficult to sent content to go through the bowel and makes it need more time for defecation during this process whether AQPs have alterdeand take part in the development and progression of STC.There is little research about that.AQPs is a kind of major intrinsic protein;More and more researches on many systems found that the pathophysiology process of many diseases had related to AQPs,including cataract,cerebral dedma,nephrogenic diabetesinsipidus (NDI),Sjogren syndrom e,pneamonedema,hepatocirrhosis et al.So it provided new target spot for treatment of some diseases.some related researches revealed that there was AQP3,4,8 in colon and had close relationship with absorption and exudation of water in colon, but that expression in colon of patients with STC is not clear.By inducing STC model and using RT-PCR technique to detect the expression of AQP3,4,8 in colonic mucosa of rats to explore the relationship between AQPs and STC。Materia and methodsThirty-two rats weighted about 150g were provided by laboratory animal research center of ZhengZhou university then they were randomly divided into two groups each group had 16 rats.Certification of fitness of that animals were SCXK:2005-001.and their weight, sex,had no significant difference, all therats were feeded in single-caged.After the purchase of the rats.there were seven days for threats to fit in with environment,then,gave the rats distilled water 0.32ml/kg BW for five days to make the rats fit the progress of lavage,then,the trial began,the rats of trial group were given distilled water 0.32ml/kg,and the rats of control group were given the solution of compound Diphenoxylate. Then the time of the black stool which was first discharged was recorded every five days On the day before this trial,and the rats of both groups were not given food except water for 12 hours,and on the next day,before this record the,rats of trial group were given were given distilled water and the rats of control group were given the solution of compound Dlphenoxylate.Thirty minutes later,both groups were given prepared Chinese into mark the time of transit progress and then both groups were given food and water normally.The time was recorded from the givings until the defecation of the first black stool,and the data was dealed with by statistical analysis, and when there were significant difference.Giving drug to the rats of control group would be stopped,After seven days,the trial was done again,and the data was analyzed again and there were also significant difference, and now,the slow transit constipation model was induced successfully.After being fasted for 12 hours,the rats were anaesthetized by 3% napental.From the abdominal incision,the skin of the rats, the colonic mucosa of rats were cut off and put into liquid nitrogen for cryopreservation.Fistly,all RNA of the colonic mucosa was extracted according to the explanation of Trizol’s kit,and the cDNA which had been synthesized was amplificated through PCR.The product was put on agarose gel for electrophoresis, the image stips of the product were scanned by High Performance Ultraviolet Trans illuminator (UVP).Their gray scale was tested by Image-Pro Plus and to (3-actin as their interior reference were relative quatity of AQP3,4,8Results1. There were expression of AQP3 in colonic mucosa of rats of both groups; RT-PCR revealed that the average value of gray scale ratios of AQP3 in proximal and distal colon of trial group and control group were 0.344 and 0.6202 (P<0.05)、0.419 and 0.509(P>0.05)respectively.2. There were expression of AQP4 in colonic mucosa of rats of both groups and the expressions of AQP4 in proximal and distal colon of trial group and control group were 0.776 and 0.736 (P>0.05)、0.776 and 0.736(P>0.05) respectively.3. There was little expression of AQP8 in both proximal and distal colon of trial and control groupsconclusion1. There was expression of AQP3 in proximal and distal colon and the expression of AQP3 jin control groups was down-regulated,there were no significant difference in both distal colon of both groups which revealed that AQP3 played an important role in water absorption which occurred in proximal colon and it may play a role in the dcvelopment and progression of STC2. There was expression of AQP4 in colonic mucosa of rats of both groups, but there was no significant difference between them,which revealed that AQP4 don’t paly the role in water absorption in colon.3. There was little expression of AQP8 in colon which revealed that AQP8 don’t paly the role in water absorption in colon.

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