中国比较医学杂志
中國比較醫學雜誌
중국비교의학잡지
CHINESE JOURNAL OF COMPARATIVE MEDICINE
2015年
3期
48-52
,共5页
螺旋涂布%Ames%联合诱导%S9%酶保护剂
螺鏇塗佈%Ames%聯閤誘導%S9%酶保護劑
라선도포%Ames%연합유도%S9%매보호제
Spiral coating%Ames%Combination inducing%S9%Enzyme protecting agent
目的：建立螺旋涂布法，评价本实验室用联合诱导法制备大鼠肝匀浆S9活性优劣，以期大大降低实验成本，并用Ames实验法评价所建立的S9低温保存方法的可行性，提高S9利用率。方法采用螺旋涂布法和Ames试验方法对市售和自制两种来源S9活化的阳性结果进行比较，并加入甘油保护剂，建立S9酶系统低温储存方法，使其以液态保存于－20℃冰箱中。结果 Ames实验中市售和自制两种S9在阳性组中都显示出明显活化作用，同一条件下，Ames试验中使用两种S9所得阳性结果并无较大差异，变率间差异无统计学意义。螺旋涂布法中，同一条件下，使用两种S9所得阳性结果也无较大差异，突变率间差异无统计学意义， S9（浓度为38％）加入量在1.48～6.62μL／μL菌液剂量下，对TA100菌有明显诱导作用，回复突变率均为对照组的3倍以上。结论可以用螺旋涂布法成功评价大鼠肝S9活性大小，联合诱导法可以替代多氯联苯诱导法制备肝匀浆S9。
目的：建立螺鏇塗佈法，評價本實驗室用聯閤誘導法製備大鼠肝勻漿S9活性優劣，以期大大降低實驗成本，併用Ames實驗法評價所建立的S9低溫保存方法的可行性，提高S9利用率。方法採用螺鏇塗佈法和Ames試驗方法對市售和自製兩種來源S9活化的暘性結果進行比較，併加入甘油保護劑，建立S9酶繫統低溫儲存方法，使其以液態保存于－20℃冰箱中。結果 Ames實驗中市售和自製兩種S9在暘性組中都顯示齣明顯活化作用，同一條件下，Ames試驗中使用兩種S9所得暘性結果併無較大差異，變率間差異無統計學意義。螺鏇塗佈法中，同一條件下，使用兩種S9所得暘性結果也無較大差異，突變率間差異無統計學意義， S9（濃度為38％）加入量在1.48～6.62μL／μL菌液劑量下，對TA100菌有明顯誘導作用，迴複突變率均為對照組的3倍以上。結論可以用螺鏇塗佈法成功評價大鼠肝S9活性大小，聯閤誘導法可以替代多氯聯苯誘導法製備肝勻漿S9。
목적：건립라선도포법，평개본실험실용연합유도법제비대서간균장S9활성우렬，이기대대강저실험성본，병용Ames실험법평개소건립적S9저온보존방법적가행성，제고S9이용솔。방법채용라선도포법화Ames시험방법대시수화자제량충래원S9활화적양성결과진행비교，병가입감유보호제，건립S9매계통저온저존방법，사기이액태보존우－20℃빙상중。결과 Ames실험중시수화자제량충S9재양성조중도현시출명현활화작용，동일조건하，Ames시험중사용량충S9소득양성결과병무교대차이，변솔간차이무통계학의의。라선도포법중，동일조건하，사용량충S9소득양성결과야무교대차이，돌변솔간차이무통계학의의， S9（농도위38％）가입량재1.48～6.62μL／μL균액제량하，대TA100균유명현유도작용，회복돌변솔균위대조조적3배이상。결론가이용라선도포법성공평개대서간S9활성대소，연합유도법가이체대다록련분유도법제비간균장S9。
Objective To reduce experimental costs and improve the utilization of S9, we use spiral coating technique to evaluate the activity of rat liver S9 prepared by combination inducing method as well as to establish cryopreservation method.Methods Using spiral coating technique and Ames test to evaluate the activity of self-made rat liver S9 and commercially available S9 separately.We use glycerinum as protective agent to establish cryogenic storage method, so that S9 can be in liquid form stored at -20 °C in the refrigerator.Results In the Ames assay as well as using spiral coating technique, the number of revertant colonies had dose-response relationship among the dose of S9.When conditions were the same, the number of revertant colonies in positive control was at the approximate level in presense of self-made rat liver S9 and commercially available S9 respectively.When S9 ( concentration of 38%) was added to the amount at 1.48 ~6.62 μL /μL broth dose of bacteria, it can significantly induced Salmonella typhimurium histidine strains TA100, reverse mutation rates were three times more than the control group.Conclusions Spiral coating technique can successfully evaluate the activity of rat liver S9.The inducing method of combination of PB and BF can take the place of the unducing method of PCBs in the preparation of Liver homogenate S9.
