CAJ | 학술논문

目的研究模拟失重环境下大鼠胰腺组织HSP70表达的变化.方法健康雄性Wistar 大鼠64只,采用尾悬吊法建立模拟失重动物模型.按完全随机法分为悬尾6、12 h及1、2、3、5、7d组和未悬尾对照组.应用免疫组化法、蛋白质印迹法和RT-PCR法检测各组大鼠胰腺组织HSP70蛋白和mRNA的表达.结果免疫组化染色显示,对照组大鼠胰腺组织无HSP70蛋白表达,悬尾组大鼠胰腺组织均有HSP70蛋白表达,尤其是胰岛细胞的胞质和胞核在早期即深染.蛋白质印迹法结果显示,悬尾6、12 h及1、2d组大鼠胰腺组织HSP70蛋白持续高表达,分别为0.921±0.078、0.862±0.048、0.838±0.063、0.807±0.071,较对照组的0.693 ±0.055显著增加(P＜0.05);3、5、7d组的HSP70蛋白表达恢复到对照组水平.HSP70 mRNA表达在6h组即明显增加至0.881 ±0.013,然后逐渐回落,3d组降至0.694±0.027,5d组出现回升,达0.836±0.014,7d组又回落至0.674±0.013.结论模拟失重使大鼠胰腺组织中HSP70蛋白和mRNA表达发生明显变化,早期过表达,后期恢复正常水平.
목적 연구모의실중배경하대서이선조직HSP70표체적변화.방법 건강웅성Wistar 대서64지,채용미현조법건립모의실중동물모형.안완전수궤법분위현미6、12 h급1、2、3、5、7d조화미현미대조조.응용면역조화법、단백질인적법화RT-PCR법검측각조대서이선조직HSP70단백화mRNA적표체.결과 면역조화염색현시,대조조대서이선조직무HSP70단백표체,현미조대서이선조직균유HSP70단백표체,우기시이도세포적포질화포핵재조기즉심염.단백질인적법결과현시,현미6、12 h급1、2d조대서이선조직HSP70단백지속고표체,분별위0.921±0.078、0.862±0.048、0.838±0.063、0.807±0.071,교대조조적0.693 ±0.055현저증가(P＜0.05);3、5、7d조적HSP70단백표체회복도대조조수평.HSP70 mRNA표체재6h조즉명현증가지0.881 ±0.013,연후축점회락,3d조강지0.694±0.027,5d조출현회승,체0.836±0.014,7d조우회락지0.674±0.013.결론 모의실중사대서이선조직중HSP70단백화mRNA표체발생명현변화,조기과표체,후기회복정상수평.
Objective To investigate Hsp70 expression features of rat's pancreas under the condition of simulated weightlessness.Methods Tail-suspension method was used to simulate the weightlessness animal model.Sixty four adult male Wistar rats were randomly divided into 8 experimental groups (n =8):suspended for 6 h,12 h,1 d,2 d,3 d,5 d,7 d and control group.Hsp70 protein and mRNA in rat's pancreas were detected by using immunohistochemical assay,Western blot and RT-PCR techniques.Results The immunohistochemistry assay showed that Hsp70 protein was expressed in rats with tail-suspension,in particular the nuclei and cytoplasm of pancreatic islet cells were deeply stained in the early phase of suspension,but was not expressed in rats without tail-suspension.The expressions of Hsp70 protein were 0.881 ±0.013,0.874 ±0.014,0.865 ±0.014,0.837 ±0.012 in 6 h,12 h,1 d,2 d groups,which were significantly higher than those in control group (0.921 ±0.078,P＜0.05),and 0.862 ±0.042,0.838 ±0.063,0.807 ±0.071 in 3 d,5 d,7 d groups,which returned to the level of control group.The expressions of Hsp70 mRNA significantly increased at 6 h,and then gradually decreased,at 3 d they reached the lowest point (0.694 ±0.027),and they began to increase to 0.836±0.014 at 5 d and decrease to 0.674±0.013 at 7 d.Conclusions The simulated weightlessness has remarkable effect on the expressions of Hsp70 protein and mRNA in rat's pancreas,and the expressions are up-regulated in the early phase and returns to normal level in the late phase.