中华医学美学美容杂志
中華醫學美學美容雜誌
중화의학미학미용잡지
CHINESE JOURNAL OF MEDICAL AESTHETICS AND COSMETOLOGY
2012年
3期
208-211
,共4页
人黑素细胞%p53%顺式咪唑啉衍生物-3%p53抑制剂%DNA氧化应激损伤
人黑素細胞%p53%順式咪唑啉衍生物-3%p53抑製劑%DNA氧化應激損傷
인흑소세포%p53%순식미서람연생물-3%p53억제제%DNA양화응격손상
Human melanocyte%p53%Nutlin-3%p53 inhibitor (PFT-α)%DNA oxidative damage
目的 通过紫外线照射(UVR)或过氧化氢(H2O2)对人黑素细胞p53的表达影响,并使用p53激活剂顺式咪唑啉衍生物-3(nutlin-3)和p53抑制剂(PFT-α)对人黑素细胞UVR或H2O2的氧化应激脱氧核糖核酸(DNA)损伤的影响,探讨p53在人黑素细胞对抗氧化应激中的作用.方法 Western印迹法测定UVR、不同浓度H2O2、nutlin-3和PFT-α处理后的人黑素细胞p53表达；单细胞电泳实验(彗星分析)测定nutlin-3或PFT-α对人黑素细胞UVR或H2O2氧化应激DNA损伤的影响；γ-H2AX免疫荧光实验测定nutlin-3对人黑素细胞UVR氧化应激DNA损伤的影响.结果 Western印迹结果显示UVR、H2O2可以增加人黑素细胞总p53的表达,并且是伴随着15丝氨酸磷酸化p53的增高而增高,nutlin-3和PFT- α分别增加和减少人黑素细胞内p53表达；彗星分析显示,预先使用nutlin-3可以明显减少UVR或H2O2造成的DNA损伤的尾素,PFT- α明显增加UVR或H2O2造成的DNA损伤的尾素,差异具有统计学意义(P＜0.05)；流式细胞仪分析显示预先使用nutlin-3可以减少UVR造成的γ-H2AX表达.结论 p53在人黑素细胞对抗UV或H2O2诱导的氧化应激中起着重要的作用.
目的 通過紫外線照射(UVR)或過氧化氫(H2O2)對人黑素細胞p53的錶達影響,併使用p53激活劑順式咪唑啉衍生物-3(nutlin-3)和p53抑製劑(PFT-α)對人黑素細胞UVR或H2O2的氧化應激脫氧覈糖覈痠(DNA)損傷的影響,探討p53在人黑素細胞對抗氧化應激中的作用.方法 Western印跡法測定UVR、不同濃度H2O2、nutlin-3和PFT-α處理後的人黑素細胞p53錶達；單細胞電泳實驗(彗星分析)測定nutlin-3或PFT-α對人黑素細胞UVR或H2O2氧化應激DNA損傷的影響；γ-H2AX免疫熒光實驗測定nutlin-3對人黑素細胞UVR氧化應激DNA損傷的影響.結果 Western印跡結果顯示UVR、H2O2可以增加人黑素細胞總p53的錶達,併且是伴隨著15絲氨痠燐痠化p53的增高而增高,nutlin-3和PFT- α分彆增加和減少人黑素細胞內p53錶達；彗星分析顯示,預先使用nutlin-3可以明顯減少UVR或H2O2造成的DNA損傷的尾素,PFT- α明顯增加UVR或H2O2造成的DNA損傷的尾素,差異具有統計學意義(P＜0.05)；流式細胞儀分析顯示預先使用nutlin-3可以減少UVR造成的γ-H2AX錶達.結論 p53在人黑素細胞對抗UV或H2O2誘導的氧化應激中起著重要的作用.
목적 통과자외선조사(UVR)혹과양화경(H2O2)대인흑소세포p53적표체영향,병사용p53격활제순식미서람연생물-3(nutlin-3)화p53억제제(PFT-α)대인흑소세포UVR혹H2O2적양화응격탈양핵당핵산(DNA)손상적영향,탐토p53재인흑소세포대항양화응격중적작용.방법 Western인적법측정UVR、불동농도H2O2、nutlin-3화PFT-α처리후적인흑소세포p53표체；단세포전영실험(혜성분석)측정nutlin-3혹PFT-α대인흑소세포UVR혹H2O2양화응격DNA손상적영향；γ-H2AX면역형광실험측정nutlin-3대인흑소세포UVR양화응격DNA손상적영향.결과 Western인적결과현시UVR、H2O2가이증가인흑소세포총p53적표체,병차시반수착15사안산린산화p53적증고이증고,nutlin-3화PFT- α분별증가화감소인흑소세포내p53표체；혜성분석현시,예선사용nutlin-3가이명현감소UVR혹H2O2조성적DNA손상적미소,PFT- α명현증가UVR혹H2O2조성적DNA손상적미소,차이구유통계학의의(P＜0.05)；류식세포의분석현시예선사용nutlin-3가이감소UVR조성적γ-H2AX표체.결론 p53재인흑소세포대항UV혹H2O2유도적양화응격중기착중요적작용.
Objective To study the effects of UVR or H2O2 on the expression of p53 in human melanocytes,and that of nutlin-3 and PFT-α on the DNA oxidative damage,and to investigate the role of p53 in the antioxidative stress.Methods The effect of UVR,H2O2,nutlin-3 and PFT-α on the expression of p53 of human melanocytes was detected by Western blot analysis,and that of nutlin-3 and PFT-α on UVR or H2O2 DNA damage assessed by single cell electrophoresis (comet assay).Determination of the effect of nutlin-3 on H2O2 DNA damage was detected by γ-H2AX immunofluorescence.Results UVR and H2O2 could induce p53 protein expression,accompanied by increased phosphorylation of p53 on serine 15 residue,and nutlin-3 and PFT-α could induce and inhibit p53 protein in human melanocytes respectively; nutlin-3 decreased the tail moment of DNA oxidative damage of UVR or H2O2 in human melanocytes,but PFT-α increased the tail moment of DNA oxidative damage of UVR or H2O2 in human melanocytes,and there were significant differences among the control and exposed groups; nutlin-3 decreased expression of γ-H2AX.Conclusions p53 plays a very important role in the antioxidative stress in melanocyte exposed to UV or H2O2.
