中华小儿外科杂志
中華小兒外科雜誌
중화소인외과잡지
CHINESE JOURNAL OF PEDIATRIC SURGERY
2014年
12期
890-894
,共5页
唐诗鹏%俞松%闫陶然%陆建国%卓金伟%柳望舒%于怀景
唐詩鵬%俞鬆%閆陶然%陸建國%卓金偉%柳望舒%于懷景
당시붕%유송%염도연%륙건국%탁금위%류망서%우부경
血管瘤%内皮,血管%基因表达调控
血管瘤%內皮,血管%基因錶達調控
혈관류%내피,혈관%기인표체조공
Hemangioma%Endothelium,vascular%Gene expression regulation
目的 观察MEK、ERK及p-ERK在体外培养血管瘤内皮细胞中的表达,探讨MEK/ERK信号通路在小儿血管瘤的发生、发展及消退过程中的作用.方法 用组织块结合酶消化法原代培养血管瘤内皮细胞,待细胞处于对数生长期,换无血清培养基培养48 h,使细胞同步化,然后加入0.1 ng/L雌二醇,50μmol/L普萘洛尔作为干预组,加入完全内皮细胞培养基作为对照组继续孵育24 h.采用Western blot法检测血管瘤内皮细胞MEK、ERK及p-ERK的表达水平,同期用流式细胞计数仪检测血管瘤内皮细胞细胞周期及凋亡的变化,并分析MEK、ERK及p-ERK的表达水平与血管瘤内皮细胞周期及凋亡的关系.结果 雌二醇干预后,MEK (0.14±0.01)、ERK1/2 (1.36±0.11/0.52±0.10)和p-ERK1/2 (0.12±0.03/0.85±0.10)与对照组MEK (0.08±0.00)、ERK1/2(1.16±0.06/0.29±0.04)和p-ERK1/2 (0.06±0.02/0.42±0.08)比较,差异均有统计学意义(t=10.01,2.86/3.45,3.57/5.95,P＜0.05);G0/G1期比例为(90.52±1.28)％,较对照组(94.02±0.85)％明显降低,差异有统计学意义(t=-3.98,P＜0.05);细胞总凋亡率为(0.70±0.10)％,较对照组(1.77±0.21)％明显降低,差异有统计学意义(t=-8.00,P＜0.05).普萘洛尔干预后,MEK(0.06±0.01)、ERK1/2 (0.87±0.05/0.14±0.02)和p-ERK1/2 (0.02±0.01/0.11±0.05)与对照组各项比较,差异均有统计学意义(t=-2.88,-6.79/-6.20,-3.96/-5.74,P＜0.05);G0/G1期比率为(96.42±1.16)％,较对照组明显升高,差异有统计学意义(t=-2.89,P＜0.05);细胞总凋亡率为(2.87±0.57)％,对照组明显升高,差异有统计学意义(t=3.14,P＜0.05).结论 MEK/ERK信号通路可能参与调控体外培养血管瘤内皮细胞的细胞周期和凋亡.雌激素、普萘洛尔可能通过干预MEK/ERK信号通路来调控体外培养血管瘤内皮细胞的增殖与凋亡.
目的 觀察MEK、ERK及p-ERK在體外培養血管瘤內皮細胞中的錶達,探討MEK/ERK信號通路在小兒血管瘤的髮生、髮展及消退過程中的作用.方法 用組織塊結閤酶消化法原代培養血管瘤內皮細胞,待細胞處于對數生長期,換無血清培養基培養48 h,使細胞同步化,然後加入0.1 ng/L雌二醇,50μmol/L普萘洛爾作為榦預組,加入完全內皮細胞培養基作為對照組繼續孵育24 h.採用Western blot法檢測血管瘤內皮細胞MEK、ERK及p-ERK的錶達水平,同期用流式細胞計數儀檢測血管瘤內皮細胞細胞週期及凋亡的變化,併分析MEK、ERK及p-ERK的錶達水平與血管瘤內皮細胞週期及凋亡的關繫.結果 雌二醇榦預後,MEK (0.14±0.01)、ERK1/2 (1.36±0.11/0.52±0.10)和p-ERK1/2 (0.12±0.03/0.85±0.10)與對照組MEK (0.08±0.00)、ERK1/2(1.16±0.06/0.29±0.04)和p-ERK1/2 (0.06±0.02/0.42±0.08)比較,差異均有統計學意義(t=10.01,2.86/3.45,3.57/5.95,P＜0.05);G0/G1期比例為(90.52±1.28)％,較對照組(94.02±0.85)％明顯降低,差異有統計學意義(t=-3.98,P＜0.05);細胞總凋亡率為(0.70±0.10)％,較對照組(1.77±0.21)％明顯降低,差異有統計學意義(t=-8.00,P＜0.05).普萘洛爾榦預後,MEK(0.06±0.01)、ERK1/2 (0.87±0.05/0.14±0.02)和p-ERK1/2 (0.02±0.01/0.11±0.05)與對照組各項比較,差異均有統計學意義(t=-2.88,-6.79/-6.20,-3.96/-5.74,P＜0.05);G0/G1期比率為(96.42±1.16)％,較對照組明顯升高,差異有統計學意義(t=-2.89,P＜0.05);細胞總凋亡率為(2.87±0.57)％,對照組明顯升高,差異有統計學意義(t=3.14,P＜0.05).結論 MEK/ERK信號通路可能參與調控體外培養血管瘤內皮細胞的細胞週期和凋亡.雌激素、普萘洛爾可能通過榦預MEK/ERK信號通路來調控體外培養血管瘤內皮細胞的增殖與凋亡.
목적 관찰MEK、ERK급p-ERK재체외배양혈관류내피세포중적표체,탐토MEK/ERK신호통로재소인혈관류적발생、발전급소퇴과정중적작용.방법 용조직괴결합매소화법원대배양혈관류내피세포,대세포처우대수생장기,환무혈청배양기배양48 h,사세포동보화,연후가입0.1 ng/L자이순,50μmol/L보내락이작위간예조,가입완전내피세포배양기작위대조조계속부육24 h.채용Western blot법검측혈관류내피세포MEK、ERK급p-ERK적표체수평,동기용류식세포계수의검측혈관류내피세포세포주기급조망적변화,병분석MEK、ERK급p-ERK적표체수평여혈관류내피세포주기급조망적관계.결과 자이순간예후,MEK (0.14±0.01)、ERK1/2 (1.36±0.11/0.52±0.10)화p-ERK1/2 (0.12±0.03/0.85±0.10)여대조조MEK (0.08±0.00)、ERK1/2(1.16±0.06/0.29±0.04)화p-ERK1/2 (0.06±0.02/0.42±0.08)비교,차이균유통계학의의(t=10.01,2.86/3.45,3.57/5.95,P＜0.05);G0/G1기비례위(90.52±1.28)％,교대조조(94.02±0.85)％명현강저,차이유통계학의의(t=-3.98,P＜0.05);세포총조망솔위(0.70±0.10)％,교대조조(1.77±0.21)％명현강저,차이유통계학의의(t=-8.00,P＜0.05).보내락이간예후,MEK(0.06±0.01)、ERK1/2 (0.87±0.05/0.14±0.02)화p-ERK1/2 (0.02±0.01/0.11±0.05)여대조조각항비교,차이균유통계학의의(t=-2.88,-6.79/-6.20,-3.96/-5.74,P＜0.05);G0/G1기비솔위(96.42±1.16)％,교대조조명현승고,차이유통계학의의(t=-2.89,P＜0.05);세포총조망솔위(2.87±0.57)％,대조조명현승고,차이유통계학의의(t=3.14,P＜0.05).결론 MEK/ERK신호통로가능삼여조공체외배양혈관류내피세포적세포주기화조망.자격소、보내락이가능통과간예MEK/ERK신호통로래조공체외배양혈관류내피세포적증식여조망.
Objective To observe the expression and regulation of MEK,ERK and p-ERK in hemangioma derived endothelial cells (HemECs) and examine the effects of MEK/ERK signal pathway in different phases of infantile hemangioma.Methods HemECs were isolated from freshly resected hemangioma specimens.During logarithmic phase,culture medium was changed to serum-free and cell synchronization started within 48 hours.HemECs were treated with 50 μm/L propranolol,0.1 ng/L estradiol for treatment group and culture medium with serum for control group after 24 hours.Then the protein levels of MEK,ERK and p-ERK,cell cycle distribution and apoptosis were observed.Finally the relationship between apoptosis,cell cycle and protein levels of MEK,ERK and p-ERK was analyzed.Results After estradiol treatment for 24 hours,the protein levels of MEK (0.14 ± 0.01),ERK1/2 (1.36 ± 0.11/0.52 ± 0.10),p-ERK1/2 (0.12 ± 0.03/0.85 ± 0.10) were higher than those of control group MEK(0.08 ± 0.00),ERK1/2 (1.16 ± 0.06/0.29 ± 0.04) and p-ERK1/2 (0.06 ± 0.02/0.42 ± 0.08).And there were significant differences (t =10.01,2.86/3.45,3.57/5.95,P＜0.05).The total ratio of cells in G0/G1 phase (90.52 ± 1.28)％ was lower than that of control group (94.02 ± 0.85) ％.And there were significant differences (t =-3.98,P＜0.05).The total ratio of apoptosis cells (0.70 ± 0.10) ％ declined compared to control group (1.77 ± 0.21) ％.And the difference was statistically significant (t =-8.00,P＜0.05).After with a 24-hour treatment of propranolol,the protein levels of MEK (0.06 ± 0.01),ERK1/2(0.87 ± 0.05/0.14 ± 0.01) and p-ERK1/2 (0.02 ± 0.01/0.11 ± 0.05) were lower than those of control group.And there was significant difference(t =-2.88,-6.79/-6.20,-3.96/-5.74,P＜0.05).Cell cycle were arrested in G0/G1 phase.Compared to control group,propranolol group (96.42 ± 1.16)％ was higher than control group.And there was significant difference(t =-2.89,P＜0.05).The total ratio of apoptotic cells (2.87 ± 0.57)％ increased versus control group.And the difference was statistically significant (t =3.14,P＜0.05).Conclusions MEK/ERK pathway plays an important role in G0/G1 phase arrest and apoptosis of hemangioma vascular endothelial cells in vitro.Estradiol and propranolol may regulate the proliferation and apoptosis of HernECs through MEK/ERK signaling pathway.
