背景:应用分子生物学技术与体内/外实验动物模型结合手段,从发育学角度探讨了热休克蛋白(heat shock proteins,HSPs)与胚胎生长发育和畸形发生的内在联系,为充实和推动从分子水平揭示胚胎发育正常或异常的奥秘开辟了一条新的途径.目的:探讨高温、香烟及联合对鼠胚胎脑HSP70及蛋白质合成的影响.设计:随机对照的实验研究.地点、材料和方法:实验在新乡医学院细胞生物学教研室和实验动物中心完成.实验选用SD雄性大鼠100只,雌性大鼠200只合笼过夜,按随机抽签法将150只孕鼠分为高温和高温与香烟联合组,高温组分为37,40,41,42℃4个大组,每一大组随机分为处理时间为1,2,3,4,5 min小组,共20组;高温与香烟联合组分为37和42℃两大组,每一大组再分为皮下注射相当于0,0.5,1.0,1.5,2.0支香烟5个小组,共10组.高温组于孕鼠孕第10天进行高温处理,高温与香烟联合组于孕鼠孕第2天进行香烟处理,各组孕鼠高温处理,2 h后处死,取胎鼠全脑组织制备匀浆,测量HSP70和蛋白质含量,观察高温及高温+香烟各组胎鼠大脑蛋白质含量及诱导合成的HSP70的含量.主要观察指标:高温、高温与香烟联合对胎鼠脑蛋白质及HSP70变化的影响.结果:高温(40,41,42℃)导致胎脑蛋白质含量从7.01 mg下降到5.66mg,与对照组(37℃)比较,差异有显著性意义(F=3.92,P＜0.01),并与温度持续时间呈正相关;同时可诱导HSP70的合成,40,41℃及42℃最高诱导量分别为57.3,71.3,83.2 mg/g,41℃持续3 min以上组及42℃各组与对照组比较有显著性差异(F=4.02,P＜0.01);高温+香烟(1.0,1.5和2.0支)组胎脑蛋白质含量分别降到5.31,5.24,5.09mg,与单独高温组比较(42℃持续5 min)胎脑蛋白质含量有明显变化(F=4.45,P＜0.01);诱导HSP70合成量与单独高温组比较未见明显变化(F=1.05,P＞0.05).结论:高温、香烟烟雾及其联合可引起胎脑蛋白质含量减少,并诱导脑HSP70合成,但香烟的诱导作用较高温弱,与高温联合胎脑HSP70合成表现出交叉适应性反应.
揹景:應用分子生物學技術與體內/外實驗動物模型結閤手段,從髮育學角度探討瞭熱休剋蛋白(heat shock proteins,HSPs)與胚胎生長髮育和畸形髮生的內在聯繫,為充實和推動從分子水平揭示胚胎髮育正常或異常的奧祕開闢瞭一條新的途徑.目的:探討高溫、香煙及聯閤對鼠胚胎腦HSP70及蛋白質閤成的影響.設計:隨機對照的實驗研究.地點、材料和方法:實驗在新鄉醫學院細胞生物學教研室和實驗動物中心完成.實驗選用SD雄性大鼠100隻,雌性大鼠200隻閤籠過夜,按隨機抽籤法將150隻孕鼠分為高溫和高溫與香煙聯閤組,高溫組分為37,40,41,42℃4箇大組,每一大組隨機分為處理時間為1,2,3,4,5 min小組,共20組;高溫與香煙聯閤組分為37和42℃兩大組,每一大組再分為皮下註射相噹于0,0.5,1.0,1.5,2.0支香煙5箇小組,共10組.高溫組于孕鼠孕第10天進行高溫處理,高溫與香煙聯閤組于孕鼠孕第2天進行香煙處理,各組孕鼠高溫處理,2 h後處死,取胎鼠全腦組織製備勻漿,測量HSP70和蛋白質含量,觀察高溫及高溫+香煙各組胎鼠大腦蛋白質含量及誘導閤成的HSP70的含量.主要觀察指標:高溫、高溫與香煙聯閤對胎鼠腦蛋白質及HSP70變化的影響.結果:高溫(40,41,42℃)導緻胎腦蛋白質含量從7.01 mg下降到5.66mg,與對照組(37℃)比較,差異有顯著性意義(F=3.92,P＜0.01),併與溫度持續時間呈正相關;同時可誘導HSP70的閤成,40,41℃及42℃最高誘導量分彆為57.3,71.3,83.2 mg/g,41℃持續3 min以上組及42℃各組與對照組比較有顯著性差異(F=4.02,P＜0.01);高溫+香煙(1.0,1.5和2.0支)組胎腦蛋白質含量分彆降到5.31,5.24,5.09mg,與單獨高溫組比較(42℃持續5 min)胎腦蛋白質含量有明顯變化(F=4.45,P＜0.01);誘導HSP70閤成量與單獨高溫組比較未見明顯變化(F=1.05,P＞0.05).結論:高溫、香煙煙霧及其聯閤可引起胎腦蛋白質含量減少,併誘導腦HSP70閤成,但香煙的誘導作用較高溫弱,與高溫聯閤胎腦HSP70閤成錶現齣交扠適應性反應.
배경:응용분자생물학기술여체내/외실험동물모형결합수단,종발육학각도탐토료열휴극단백(heat shock proteins,HSPs)여배태생장발육화기형발생적내재련계,위충실화추동종분자수평게시배태발육정상혹이상적오비개벽료일조신적도경.목적:탐토고온、향연급연합대서배태뇌HSP70급단백질합성적영향.설계:수궤대조적실험연구.지점、재료화방법:실험재신향의학원세포생물학교연실화실험동물중심완성.실험선용SD웅성대서100지,자성대서200지합롱과야,안수궤추첨법장150지잉서분위고온화고온여향연연합조,고온조분위37,40,41,42℃4개대조,매일대조수궤분위처리시간위1,2,3,4,5 min소조,공20조;고온여향연연합조분위37화42℃량대조,매일대조재분위피하주사상당우0,0.5,1.0,1.5,2.0지향연5개소조,공10조.고온조우잉서잉제10천진행고온처리,고온여향연연합조우잉서잉제2천진행향연처리,각조잉서고온처리,2 h후처사,취태서전뇌조직제비균장,측량HSP70화단백질함량,관찰고온급고온+향연각조태서대뇌단백질함량급유도합성적HSP70적함량.주요관찰지표:고온、고온여향연연합대태서뇌단백질급HSP70변화적영향.결과:고온(40,41,42℃)도치태뇌단백질함량종7.01 mg하강도5.66mg,여대조조(37℃)비교,차이유현저성의의(F=3.92,P＜0.01),병여온도지속시간정정상관;동시가유도HSP70적합성,40,41℃급42℃최고유도량분별위57.3,71.3,83.2 mg/g,41℃지속3 min이상조급42℃각조여대조조비교유현저성차이(F=4.02,P＜0.01);고온+향연(1.0,1.5화2.0지)조태뇌단백질함량분별강도5.31,5.24,5.09mg,여단독고온조비교(42℃지속5 min)태뇌단백질함량유명현변화(F=4.45,P＜0.01);유도HSP70합성량여단독고온조비교미견명현변화(F=1.05,P＞0.05).결론:고온、향연연무급기연합가인기태뇌단백질함량감소,병유도뇌HSP70합성,단향연적유도작용교고온약,여고온연합태뇌HSP70합성표현출교차괄응성반응.
BACKGROUND:To discuss the interrelations between heat shock proteins (HSPs) and the growth development and teratogenesis of embryo from the auxological angle by combining the molecular biological skills and in vivo/in vitro experimental animal models in order to discover the reasons of normal or abnormal development of embryo from the molecular level.OBJECTIVE:To explore the effects of hyperthermia, cigarette to the synthesis of HSP70 and proteins of rat embryo.DESIGN:Randomized case controlled study.SETTING, PARTICIPANTS and INTERVENTIONS:Experiment was done in the Biology Teaching Section and Experimental Animal Center of Xinxiang Medical College.The 100 male SD rats and 200 female rats were put into same cages.150 pregnant rats were divided into high temperature groups and combining high temperature and cigarette groups.High temperature groups included 37, 40, 41, 42 ℃ groups and each was divided into 5sub-groups with 1, 2, 3, 4, 5 minutes of treat time respectively, totally 20groups.The combining groups of high temperature and cigarette were divided into 37 ℃ group and 42 ℃ group, each was divided into 5 sub-groups with subcutaneously injected 0, 0.5, 1.0, 1.5 and 2 cigarettes, totally 10groups.High temperature groups were processed in the tenth day of pregnancy, combining groups were processed the cigarette treatment in the 2 days of pregnancy.The rats of all the groups were executed in the 2 hours after high temperature treatment.Whole brain of rat embryo was used to make homogenate and then HSP70 and content of proteins were assayed.MAIN OUTCOME MEASURES:Changes of brain proteins and HSP70 of rats' embryo with high temperature treatment and combining treatment of high temperature and cigarette.RESULTS:The content of embryo brain protein decreased from 7.01 mg to 5.66 mg after high temperature treatment(40, 41, 42 ℃), there was significant difference between these groups and control group(37 ℃) (F=3.92, P ＜0.01) .There was positive correlation between the content and lasting time of high temperature.The synthesis of HSP70 was induced to produce, the highest inductive amount of 40, 41, 42 ℃ was 57.3, 71.3 and 88.2 mg/g respectively.There was significant difference between 41 ℃ groups with lasting time more than 3 minutes, 42 ℃ groups and control group( F=4.02, P ＜ 0.01) .The embryo brain proteins of combining group with 1, 1.5 and 2 cigarettes reduced to 5.31, 5.24, 5.09 mg respectively, there was difference between these groups and solo high temperature groups(42 ℃ for 5 minutes)(F = 4.45, P ＜0.01) .There was no obvious difference on the amount of HSP70 between combining groups and solo high temperature groups( P ＞ 0.05).CONCLUSION:High temperature, smoke of cigarettes as well as the combining effects of both can lead to decreased contents of embryo brain protein and induce the synthesis of HSP70.But the inductive action of cigarette is lower than that of high temperature.There is cross adaptative reaction on synthesis of HSP70 when combining with high temperature.
