临床口腔医学杂志
臨床口腔醫學雜誌
림상구강의학잡지
JOURNAL OF CLINICAL STOMATOLOGY
2015年
8期
451-455
,共5页
低氧%低氧诱导因子1α%过氧化物酶体增殖物激活受体γ协同刺激因子1β%AMP依赖的蛋白激酶
低氧%低氧誘導因子1α%過氧化物酶體增殖物激活受體γ協同刺激因子1β%AMP依賴的蛋白激酶
저양%저양유도인자1α%과양화물매체증식물격활수체γ협동자격인자1β%AMP의뢰적단백격매
hypoxia%hypoxia inducible factor-1α(HIF-1α)%peroxisome proliferatoractivated receptor gamma coacti-vator-1β(PGC-1β)%AMP-activated protein kinase(AMPK)
目的：研究低氧诱导因子HIF－1α在颏舌肌成肌分化和线粒体生物发生中的作用，探索可能的调控机制。方法：构建HIF－1α敲降质粒，慢病毒载体转染小鼠颏舌肌成肌细胞，设阴性对照组，用含2％马血清的分化培养基在低氧（1％O2）环境下诱导成肌细胞分化，western blot检测成肌细胞分化2、4、6 d，PGC－1β、AMPKα1、pAMPKα1的表达水平，免疫细胞化学染色观察低氧分化4 d时AMPK通道激活剂与阻滞剂对PGC－1β的表达影响，western blot定量分析。结果：①HIF－1α敲降组的PGC－1β表达量高于对照组（P＜0．05），且分化4 d时表达最高（P＜0．05）；②AMPKα1总蛋白表达量无明显差异，pAMPKα1的表达在HIF－1α敲降组远高于对照组（P＜0．05）。③AMPK通路促进剂AICAR增加PGC－1β的表达（P＜0．05），抑制剂Compound C明显抑制PGC－1β的表达（P＜0．05）。结论：HIF－1α下调了低氧环境中PGC－1β的表达，PGC－1β的表达受AMPK通路的正向调节作用。
目的：研究低氧誘導因子HIF－1α在頦舌肌成肌分化和線粒體生物髮生中的作用，探索可能的調控機製。方法：構建HIF－1α敲降質粒，慢病毒載體轉染小鼠頦舌肌成肌細胞，設陰性對照組，用含2％馬血清的分化培養基在低氧（1％O2）環境下誘導成肌細胞分化，western blot檢測成肌細胞分化2、4、6 d，PGC－1β、AMPKα1、pAMPKα1的錶達水平，免疫細胞化學染色觀察低氧分化4 d時AMPK通道激活劑與阻滯劑對PGC－1β的錶達影響，western blot定量分析。結果：①HIF－1α敲降組的PGC－1β錶達量高于對照組（P＜0．05），且分化4 d時錶達最高（P＜0．05）；②AMPKα1總蛋白錶達量無明顯差異，pAMPKα1的錶達在HIF－1α敲降組遠高于對照組（P＜0．05）。③AMPK通路促進劑AICAR增加PGC－1β的錶達（P＜0．05），抑製劑Compound C明顯抑製PGC－1β的錶達（P＜0．05）。結論：HIF－1α下調瞭低氧環境中PGC－1β的錶達，PGC－1β的錶達受AMPK通路的正嚮調節作用。
목적：연구저양유도인자HIF－1α재해설기성기분화화선립체생물발생중적작용，탐색가능적조공궤제。방법：구건HIF－1α고강질립，만병독재체전염소서해설기성기세포，설음성대조조，용함2％마혈청적분화배양기재저양（1％O2）배경하유도성기세포분화，western blot검측성기세포분화2、4、6 d，PGC－1β、AMPKα1、pAMPKα1적표체수평，면역세포화학염색관찰저양분화4 d시AMPK통도격활제여조체제대PGC－1β적표체영향，western blot정량분석。결과：①HIF－1α고강조적PGC－1β표체량고우대조조（P＜0．05），차분화4 d시표체최고（P＜0．05）；②AMPKα1총단백표체량무명현차이，pAMPKα1적표체재HIF－1α고강조원고우대조조（P＜0．05）。③AMPK통로촉진제AICAR증가PGC－1β적표체（P＜0．05），억제제Compound C명현억제PGC－1β적표체（P＜0．05）。결론：HIF－1α하조료저양배경중PGC－1β적표체，PGC－1β적표체수AMPK통로적정향조절작용。
Objective:To investigate the effects of HIF-1α on myogenic differentiation and mitochondrial biogenesis and the mechanism involved. Method:HIF-1α shRNA lentivirous was used to infect the myoblasts. Myoblasts was induced differentiation by 2%horse serum in hypoxia condition(1%O2) for 6 days,PGC-1β,AMPKα1 and pAMPKα1 were detect-ed through western-blot. Immunocytochemistry and western-blot were performed to detected the expression of PGC-1β after medication of AICAR or Compound C. Result: ①Compared with control myoblasts,the expression of PGC-1βwas higher in HIF-1α KO myoblasts during myogenesis (P <0.05),and highest in the fourth day. ②There is no significant differences in AMPKα1 (P >0.05). The expression of pAMPKα1 was significantly higher under hypoxia condition in HIF-1α KO group, compared with the control groups(P <0.05).③AICAR(activator of AMPK) increaed the expression of PGC-1β(P <0.05), while Compound C (inhibitor of AMPK) showed opposite effect (P<0.05). Conclusion:HIF-1αdecreases the expression of PGC-1βunder hypoxia condition. The expression of PGC-1β is influenced by AMPK,which acts as a positively acting com-ponent.
