| ObjectiveFrom flies life cycle, Building Trpal RNAi is gene, the sports ability and cardiac function in three aspects Trpal candidate genes and evaluation in the sports of the anti-aging functional effect, for screening other candidate genes for research foundation.Metairie and methodsTrpal gene RNAi construct:take eight hours Gal4provided strain (hunan normal university heart development research center provide) male fruit flies and UAS-Trpal strain (bought in the United States Birmingham strain center) virgin flies hybrid, Fl Trpal RNAi is on behalf of genes, pick take12hours feather F12400male fruit flies only randomly divided into Trpal gene RNAi is quiet group, Trpal gene RNAi is flat in the exercise group. Another pick2400only12hours feather UAS-Trpal gene fruit flies, experimental group for UAS-Trpal gene quiet group, UAS-Trpal gene flat in the exercise group. Each group of each1200only,(breeding conditions for25℃,50%constant temperature, humidity,12/12hours a day and night light cycle).The fruit flies sports training platform,8day old executive regularity platform sports training, set:the motor speed for24s/r,9.5cm landing platform height value. Application video detection system monitoring the training intensity. Two days before the adaptability for training, the six day old adaptation20minutes,7day old for45minutes,8day old began to exercise is training, training time was2.5hours, training for the next day training a frequency, until the end of scheme.Sports proposal execution after the end of the12h randomly selected in each category respectively take fruit fly life live index, sport climbing ability, heart function test. The determination of the life live index from sports after the next day began to plan, take the group fruit flies100only, same breeding environment, the next day a new culture medium (replacement for the same formula of the medium with batch preparation), every recording the group deaths fruit fly, drosophila make live curve. Sport climbing ability test after24hours from movement began, each group of random fruit flies100only to each tube for20standard, tube specifications for high standard size=×20cm×2cm, cotton plug from the bottom to reserve18cm as climbing area, in order to stick from the bottom to the test tube division for1,2,3,4,5,6,7,8,9nine area, each area for high×size=2cm×2cm, each region of the score of the corresponding1,2,3,4,5,6,7,8,9points, through the video detection system within the time required, regional counting the number of fruit flies, the area of the fruit flies number and the area is for the region to score the sum total score, calculate the area must points for drosophila and that is to climb the index. Take each day14age group fruit flies,28day old fruit flies,42day old fruit flies the continuous only100heart function testing. The method is:pulse electrical stimulation PHIPPS&BIRD instrument, a value of40V,6Hz,30s exciting, two minutes see heart tube recovery (normal, fibrous vibrate, stop). Were counting, fibrous vibrate and stop the percentage of the total number of the fruit flies and namely for heart failure rate.ResultsRats in fruit flies Trpal genes of homologous gene search resultsBLASP through the sequence alignment, SMART structure prediction and related literatures in recent years consult, found in fruit flies species transient receptor potential A1genetic encoding protein whether level1sequence or function prediction and rats are Trpal gene protein has high similarity, so sure rats in the fruit fly species Trpal gene homology gene for transient receptor potential A1.Life-span of the values of the test results(1) in the fruit fly half time of death of detecting the UAS-Trpal gene flat in the exercise group fruit flies half life value than quiet death control group of fruit flies high value life, and has a significant difference (P0.05), a statistically significant difference.(2) the average life expectancy in fruit flies of detecting the Trpal gene RNAI flat in the exercise group fruit flies average life expectancy than RNAI quiet value life-span control group is low, and with significant difference (P<0.05), a statistically significant difference.(3) In fruit flies highest life of detecting the Trpal gene RNAI blank control group fruit flies highest value life than UAS-Trpal gene blank control group low, and with significant difference (P<0.05), Trpal gene RNAI exercise group the highest life value Trpal genes than control group, and low RNAI blank with significant difference (P<0.05), a statistically significant difference.(4) UAS-Trpal RNAi exercise group and blank compared with controls, and prolong its life increased an average rate, Hand>Trpal RNAi control group and blank compared with controls, rate and prolong its life, the sports>Trpal RNAi fruit flies Hand intervention on average after rate obviously and prolong its life.Fruit flics climbing ability test resultsAlong with the age, fruit flies climbing ability significant down which UAS-Trpal RNAi exercise group climbing ability in sports training significantly improved under that exercise training slows the fruit flies climbing ability of aging. And Trpal RNAi>Hand after the sports ability not interference than before dropped significantly, and does not help the exercise interventions to climb the level.Fruit flies heart form observationsUAS-Trpal RNAi larvae tube and Hand>heart Trpal RNAi larvae heart heart tube cells are arranged than that of rules, the heart is developing normally. Through the observation of5weeks age heart tube structure form fruit fly, drosophila UAS-Trpal RNAi tube and the heart of the Hand> Trpal RNAi heart to the form of the arrangement of the rules are more closely, myocardial cells and around the heart cells form size no obvious difference.Fruit flies heart rate failure test resultsFruit flies heart failure rate and fruit flies age with high correlation, and blank compared with controls, UAS-Trpal RNAi platform in the exercise group of heart failure rate is drosophila influence, Hand>Trpal RNAi the flies heart rate significantly increased rate of failure, sports Trpal intervention will make this gene interference lines a failure is more obvious.4. Conclusion(1) Trpal genes after RNAi can lead to fruit flies shorten life cycle;(2)2.5hours of platform movement to slow the UAS-Trpal strain of fruit flies climbing ability; Trpal RNAi genes after climbing ability fly down significantly,2.5hours of platform movement makes this a sports recession intensified;(3) Trpal RNAi genes after heart failure rate flies significantly increased,2.5hours of platform movement heart failure aggravate that Trpal gene in retarding aging flies heart plays an important role. |
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