The Change Of AQP3,8, And9in The Residual Colonic Mucosa Of Partial Resection Of Colon Of The SD Rat

Background and aimIt is reported that the Water channel protein (Aquaporin) is known as one kind of hole protein, which is located on the cell membrane.And the protein on the cell membrane, it can control the water in and out of the cells. The Water channel protein family (AQPs) widely exists in the cell membranes of the prokaryotic and eukaryotic, it also can efficiently and selectively transport the water molecules. As a huge across the membrane channel proteins (the major intrinsic protein, The MIP),the AQPs play an important role in adjusting the water metabolism of plants and animals. In1988, Agre found it and named it as CHIP. After this special protein was found, scholars foundabout13kinds of AQP (AQP0-12)in mammals through immunohistochemistry、 RT-PCR and so on. Domestic and foreign literatures reported AQP3,8,9, colon, have close relationship with colon moisture absorption and secretion. In the clinical work, to remove part of the colon in patients of colorectal cancer, and the incidence of diarrhea occurs about5～30%after surgery. And the patients need to experience a hard compensatory period, exist in the colonic mucosa of AQP compensatory, whether the same AQP is how to change. This experiment by making rat colon resection model, and detecting AQP38,9in the residual colonic mucosa, in accordance with the principle of rt-pcr. We try to find a way from micro Angle,in order to find a good way, which can improve the quality of colorectal cancer postoperative defecation train of thought, so that furtherly improve the patient’s postoperative quality of life. The SD rats with Most of the colon resection, AQPs expression how changes and takes place during this period, whether the expression changes related to compensation. Materials and methodsAbout12weeks of Sprague wrote-Dawley (SD) rats from Zhengzhou university animal center, the they were randomly divided into two groupsand each group had16.The average weight is controlled between250g and300g, male and female half, experimental animal certificate number is:SCXK (prepare)2010-0002, all the rats were randomly divided into blank control group and experimental group,16VS16; Bought in rats after experimental animal center in He Nan province are single cage, using the national standard of rat feed, breeding,7days to adapt to the environment, eliminate stress effects on gastrointestinal tract environment. Controlgroupand experimental group rats do statistical analysis of gender, age, and weight no difference, no statistical significance, are comparable,(specific see table1); statistical analysis Blank group rats don’t operation, the experimental group rats most of colon resection; Experimental group rats before surgery began to fast,24hours a day can’t help but water; Preoperative in experimental group rats weighing respectively, according to10mg/kg dose, the line of intraperitoneal injection of anesthetic rats, and then used scissors to abdominal MAO issued regular for skins and disinfection; Surgery was carried out in bench, midline incision abdomen, first find the colon, at about5cm from ileocecus most of colon resection,6/0with no wound suture continuous monolayer of end to end anastomosis bowel, then use suture mesocolon, before closing abdomen abdominal cavity injection of penicillin sodium200000units, with the ordinary silk thread finally closed abdomen, step by step of metabolic cage was put rats;24hours after the surgery, using rats lavage apparatus for postoperative perfusion glucose and sodium chloride in rats to replenish electrolytes and some nutrition, continuous two days later, the rats of normal free food, water, SD rats colon resection model is established in this paper.14days after the surgery, a fast water for12hours, random control group8rats, executed after anesthesia, capture the remaining colon and scrape mucous membrane, cryopreserved in liquid nitrogen tank waiting for inspection;28days after the surgery, put to death of the experimental group rats, remaining and extract the remaining colon mucosa, cryopreserved waiting for inspection; Blank control group rats in the group after28days, prohibit watering for12hours, all put to death, to extract the colonic mucosa of freeze-stored waiting for inspection. According to RT-PCR method, testing all SD rats in this experiment AQP3,8,9in the colonic mucosa,:first to extract total RNA, according to the instructions of TRIZOL reagent kit in the control group, postoperative group,14days after28days group, respectively To extract total RNA; Second for PCR amplification, in accordance with the related cDNA kit, and the results show that:in strict accordance with the steps of three groups of total RNA RT-PCR amplification; Third RT-PCR amplification results on AGAR candy electrophoresis, the electrophoresis results in High Performance Ultraviolet Trans illuminator (UVP) gel imaging system on the camera, and then observed after amplification bands and it fits the purpose stripe, finally using related software of electrophoretic bands of grey value determination, and gray value of the target protein made in the ratio of the grey value of protein, as statistical data of the experiments, and then use the statistics software to carry on the quantitative analysis.Results1, The experimental group rats, beginning after defecation, stool water content than the blank control group rats, significantly increased;2,2.RT-PCR revealed that, the mean gray scale ratios of aqp3,8,9in the control group,14th group and28th group were0.352.0.425、0.614(p<0.01),0.425、0.518、0.733(p<0.01),0.422、0.516、0.632(p<0.01)Conclusion1, In sd rats of most of colon resection,14th and28th day after the remaining amount of the expression of AQP3colon mucosa,8,9express increase compared with blank control group,28days after is14days after surgery group expression quantity increase further.2, after most of colon resection, residual colonic mucosa compensatory increased the expression of AQP3,8,9, which is the carrier of the remaining colon water channel protein quantity increased, and the postoperative colonic vicarious absorption has a certain relationship.