中华烧伤杂志
中華燒傷雜誌
중화소상잡지
16
2008年
6期
450-453
,共4页
李牧%王裴%刘琛%陈传莉%王凤君
李牧%王裴%劉琛%陳傳莉%王鳳君
리목%왕배%류침%진전리%왕봉군
缺氧%寡霉素类%缺氧诱导因子1α%肠上皮细胞
缺氧%寡黴素類%缺氧誘導因子1α%腸上皮細胞
결양%과매소류%결양유도인자1α%장상피세포
Hypoxia%Oligomycins%Hypoxia-inducible factor1 α%Intestinal epithelial cells
目的 了解缺氧对肠上皮细胞缺氧诱导因子1α(HIF-1α)活化的影响. 方法 将肠上皮细胞分为常氧处理(正常对照)、缺氧(设缺氧1、2、6、12、24 h)及缺氧+寡霉素处理(分别用浓度为5、10、20、40μg/mL寡霉素处理1 h,再缺氧6 h).采用蛋白质印迹法检测HIF-1α蛋白表达,免疫荧光法观察HIF-1α向细胞核转位的情况. 结果 与正常对照(0.08±0.07)相比较,缺氧1 h肠上皮细胞HIF-1α蛋白表达(0.52±0.30)即显著升高(P＜0.05),6 h达峰值(2.37±1.08,P＜0.05),同时HIF-1α向细胞核转位也明显增加.寡霉素呈剂量依赖性地抑制缺氧引起的肠上皮细胞HIF-1α蛋白表达增加,用5、10、20及40μg/,mL寡霉素处理的缺氧肠上皮细胞HIF-1α蛋白表达量分别为1.62±0.96、1.48±0.56、1.08±0.36及0.58±0.11,均较单纯缺氧6 h(2.67±1.38)显著降低(P＜0.05),HIF-1α向细胞核转位也被抑制. 结论 呼吸链抑制剂寡霉素可抑制缺氧肠上皮细胞HIF-1α活化,线粒体呼吸链可能在其发生机制中具有重要作用.
目的 瞭解缺氧對腸上皮細胞缺氧誘導因子1α(HIF-1α)活化的影響. 方法 將腸上皮細胞分為常氧處理(正常對照)、缺氧(設缺氧1、2、6、12、24 h)及缺氧+寡黴素處理(分彆用濃度為5、10、20、40μg/mL寡黴素處理1 h,再缺氧6 h).採用蛋白質印跡法檢測HIF-1α蛋白錶達,免疫熒光法觀察HIF-1α嚮細胞覈轉位的情況. 結果 與正常對照(0.08±0.07)相比較,缺氧1 h腸上皮細胞HIF-1α蛋白錶達(0.52±0.30)即顯著升高(P＜0.05),6 h達峰值(2.37±1.08,P＜0.05),同時HIF-1α嚮細胞覈轉位也明顯增加.寡黴素呈劑量依賴性地抑製缺氧引起的腸上皮細胞HIF-1α蛋白錶達增加,用5、10、20及40μg/,mL寡黴素處理的缺氧腸上皮細胞HIF-1α蛋白錶達量分彆為1.62±0.96、1.48±0.56、1.08±0.36及0.58±0.11,均較單純缺氧6 h(2.67±1.38)顯著降低(P＜0.05),HIF-1α嚮細胞覈轉位也被抑製. 結論 呼吸鏈抑製劑寡黴素可抑製缺氧腸上皮細胞HIF-1α活化,線粒體呼吸鏈可能在其髮生機製中具有重要作用.
목적 료해결양대장상피세포결양유도인자1α(HIF-1α)활화적영향. 방법 장장상피세포분위상양처리(정상대조)、결양(설결양1、2、6、12、24 h)급결양+과매소처리(분별용농도위5、10、20、40μg/mL과매소처리1 h,재결양6 h).채용단백질인적법검측HIF-1α단백표체,면역형광법관찰HIF-1α향세포핵전위적정황. 결과 여정상대조(0.08±0.07)상비교,결양1 h장상피세포HIF-1α단백표체(0.52±0.30)즉현저승고(P＜0.05),6 h체봉치(2.37±1.08,P＜0.05),동시HIF-1α향세포핵전위야명현증가.과매소정제량의뢰성지억제결양인기적장상피세포HIF-1α단백표체증가,용5、10、20급40μg/,mL과매소처리적결양장상피세포HIF-1α단백표체량분별위1.62±0.96、1.48±0.56、1.08±0.36급0.58±0.11,균교단순결양6 h(2.67±1.38)현저강저(P＜0.05),HIF-1α향세포핵전위야피억제. 결론 호흡련억제제과매소가억제결양장상피세포HIF-1α활화,선립체호흡련가능재기발생궤제중구유중요작용.
Objective To investigate the effect of hypoxia on HIF-1 α activation in intestinal epithelial cells. Methods Intestinal epithelial cells were randomly divided into normal control group,hypoxia group and hypoxia plus oligomycin group(otigomycin group).In hypoxia group,the cells were exposed to hypoxia for 1,2,6,12 and 24 h.In oligomycin group,the cells were treated with oligomycin in concentration of 5,10,20 and 40μg/mL for 1 h prior to 6-hour hypoxic exposure.HIF-1α protein expression was assayed by western blot method.Nuclear translocation of HIF-1 α was detected by immunofluorescence analysis. Results Compared with that in control group(0.08±0.07),HIF-1 α protein expression in hypoxia group increased significantly at 1 h(0.52±0.30,P＜0.05),and reached the peak value(2.37±1.08,P＜0.05)at 6 h.Nuclear transloeation of HIF-1 α was also induced by hypoxia.HIF-1 α protein expression in oligomyein group in the concentration of 5,10,20 and 40μg/mL of oligomycin was 1.62±0.96,1.48±0.56,1.08±0.36 and 0.58±0.11 respectively,which was significantly lower than that only after exposure to hypoxia for 6 h(2.67±1.38.P＜0.05).The nuclear translocation of HIF-1α induced by hypoxia was also obviously inhibited by oligomycin pretreatment. Conclusion Oligomycin,a specific inhibitor of respiratory chain,inhibits HIF-1 α.activation,which suggests that mitochondria respiratory chain may play an important role in aforementioned process.
