Study On The Relationship Between The High Temperature Line On The Experimental Model Rabbit Of Cholecystitis And The High Expression Of UCP Genes In Rabbit Adipose Tissue

Objective:To establish an experimental model rabbit of cholecystitis, which can cause the production of the high temperature line, and to explore the distrabution characters of the high temperature line, changes of the testing temperatures. To find out the relationship between the high temperature line and the high expression of UCP1 genes in rabbit adipose tissue in the production of heat of the adipose tissue, to provide the experimental foundation for the mechanism of the production of the high temperature line.Methods:The research is devided into two parts.PARTⅠ:Rabbits were devided randomly into the experimental cholecystitis model group and the control group. Experimental model rabbit of cholecystitis were induced by injecting E. Coli solution into the gallbladder of the rabbits after the operation of the narrowing of common bile duct ligation. Before the operation and 14 days after the operation, such testing indicators were measured:the pictures collected by the infrared thermal imager and temperatures measured by the infrared thermometer. Compare the two testing indicators mentioned above; Analyse the successful rates of the production of the high temperature line by building the experimental model rabbit of cholecystitis; observe the morphological characters of the high temperature line, and the changes among the testing indicators; compare the changes of the temperatures between the day before the operation and 14 days after the operations.PARTⅡ:Rabbits in the experimental cholecystitis model group in which the high temperature line was successfully produced and their compared rabbits in the control group were selected as experimental animals in this experiment. Adipose tissue was cut off on the 14th day after the operation. The total RNA was isolated from the adipose tissue by UNIQ-10 method and Trizol method. And then the comparison between the quality of the total RNA isolated by the two methods above was analysed. After the experiment of RT, the expression of UCP1 gene of the adipose tissue was determined with Real-Time PCR in order to observe the effect of the high temperature line on the expression of UCP1 genes in rabbit adipose tissue.Results:PARTⅠ:1. By comparing the pictures collected by the infrared thermal imager and the temperatures measured by the infrared thermometer, the tendency of the changes of the temperatures showed by the two measuring method mentioned above was found equal. The tendency is that the temperatures of the rabbits on the 14th day after the operation are higher than the temperatures of the rabbits before the operation, and the temperatures of the head-side of the rabbits are higher than the temperatures of the tail-side. And this result shows that although differences of acute measuring may exist to cause differences in temperature measuring results round 1℃, the tendency showed by the two methods is equal. Then the practical possibility of the infrared thermometer in the research is proved, and the testing indicators can be measured by such a method.2. By observing the high temperature line which was successfully produced by building the experimental model rabbit of cholecystitis, there are high temperature lines going along both sides of the rabbit's body. It's direction is as the same as the Foot ShaoYang meridians. Some of the high temperature lines go along with other meridians. The average lenth is 5-9 cm, and the longest can reach to both ends of the rabbit's body. The characters of the change of the temperature:the temperature of the head-side is higher than the tail-side, and the temperature of the right side is much higher than the left side.3. In the experiment in which the successful rate of the production of the high temperature line caused by the building of the experimental model rabbit of cholecystitis was analysed, the result of the analysis of the rate was calculated as 60% through the testing indicators and the analysing charts.4. In the experiment in which high temperature line was produced by building the experimental model rabbit of cholecystitis, temperatures were analysed. The result of the analyse:The temperatures of the rabbits in the experimental model group on the 14th day after the operation are higher than the temperatures measured before the operation, and the changes are significant(P<0.01); The changes of the temperatures of the rabbits in the control group are not significant(P>0.01); Compared with the temperatures of the rabbits in the control group, the difference of the temperatures between the 14th day after the operation and the day before the operation in the experimental model group is significant(P<0.01).PARTⅡ:1. In the comparison between the UNIQ-10 method and Trizol method in the isolation of the total RNA from the adipose tissue, the result of the comparison of the purity of the total RNA is that the purity of the total RNA isolated by the Trizol method (the average of OD, A260/A280 is 1.598) is higher than than the purity of the total RNA isolated by the UNIQ-10 method (the average of OD, A260/A280 is 1.562). Although the purity of the total RNA isolated by the UNIQ-10 method is lower, its purity fits the standard of the quality of the total RNA acquired in the practical experiment; the result of the comparison of the totality of the total RNA is that the totality of the total RNA isolated by the Trizol method is better than the totality of the total RNA isolated by the UNIQ-10 method. The picture of electrophoresis shows that all the belts are clear and highlight, and both the highlightness and clearness of the belts of the total RNA isolated by the Trizol method are better than those belts of the total RNA isolated by the UNIQ-10 method. Such a result indicates that though the totality of the total RNA isolated by the two methods fit the quality of the total RNA acquired in the practical experiments of RT and Real-Time PCR, the total RNA isolated by the Trizol method is better and more suitable to isolate the total RNA from the rabbits adipose tissue in this experiment.2. In the experiment of analysing the effect of the effect of the high temperature line on the expression of UCP1 genes in rabbit adipose tissue, the result of the expression of UCP1 genes in rabbit adipose tissue in the model group and the control group measured by Real-Time PCR is that comparing with the control group, Ct gets lower significantly, and the expression of UCP1 genes in rabbit adipose tissue in the model group gets higher, and the changes are significant(P<0.01).Conclusions:1. The high temperature line can be produced by building the experimental model rabbit of cholecystitis; The temperatures of the high temperature line which is produced successfully are suitable to the standard of the testing indicators and the morphological characters acquired by the practical experimental researches.2. The practical using possibility of the infrared thermometer is proved in the process of the collection of the testing indicators acquired by the practical research of the high temperature line.3. The expression of UCP1 genes in rabbit adipose tissue in the model group can be effected by the production of the high temperature line. Such a result indicates that UCP1 is an important factor in the process of producing heat in the adipose tissue, and it has an intimate relationship with the production of the high temperature line.