中华急诊医学杂志
中華急診醫學雜誌
중화급진의학잡지
CHINESE JOURNAL OF EMERGENCY MEDICINE
2009年
1期
56-59
,共4页
张万福%胡大海%吕根法%王耘川%朱雄翔%高峰
張萬福%鬍大海%呂根法%王耘川%硃雄翔%高峰
장만복%호대해%려근법%왕운천%주웅상%고봉
胰岛素%烧伤%内皮细胞%凋亡%内皮型一氧化氮合酸
胰島素%燒傷%內皮細胞%凋亡%內皮型一氧化氮閤痠
이도소%소상%내피세포%조망%내피형일양화담합산
Insulin%Bum%Endothelial cell%Apoptosis%Endothelial nitric oxide synthase(eNOS)
目的 探讨胰岛素对烧伤血清诱导血管内皮细胞凋亡的影响及相关机制.方法 体外常规培养内皮细胞系ECV304,随机分为正常对照组(n=6),体积分数15%(V/V)正常大鼠血清培养;烧伤血清刺激组(n=6),体积分数15%(V/V)自制烧伤大鼠血清培养,血清采自背部30%TBsAIII 烫伤大鼠模型;胰岛素处理组(n=6),体积分数15%(V/V)自制烧伤大鼠血清中加入10-7M/L胰岛素培养;刺激6 h后采用原位末端标记法(TUNEL)观察内皮细胞凋亡、免疫组织化学染色和蛋白印迹法枪测抗凋亡蛋白bcl-2及eNOS的蛋白表达变化,数据以表示,均数比较采用单凶素方差分析,以P<0.05为差异具有统计学意义.结果 与正常对照组比较,烧伤血清促进内皮细胞捌亡(18.5%±3.1%),免疫组化染色bcl-2平均吸光度低(0.14±0.02),蛋白印迹实验显示bel-2(0.36 4±0.12)和p-eNOS(0.55±0.28)表达降低(P<0.01).相对十烧伤血清刺激组,胰岛素处理组细胞凋亡显著减少(9.6 4±2.8%),免疫组化染色bcl-2平均吸光度升高(0.21±0.03),蛋白印迹实验显示bcl.2(0.94 4-0.25)和p-eNOS(0.89±0.16)表达明显恢复(P<0.01).eNOS在各组中无明显变化.结论 胰岛素明显抑制烧伤血清诱导下的内皮细胞凋亡并增加扰凋亡蛋白bcl-2的表达,此作用町能与eNOS的磷酸化有关.
目的 探討胰島素對燒傷血清誘導血管內皮細胞凋亡的影響及相關機製.方法 體外常規培養內皮細胞繫ECV304,隨機分為正常對照組(n=6),體積分數15%(V/V)正常大鼠血清培養;燒傷血清刺激組(n=6),體積分數15%(V/V)自製燒傷大鼠血清培養,血清採自揹部30%TBsAIII 燙傷大鼠模型;胰島素處理組(n=6),體積分數15%(V/V)自製燒傷大鼠血清中加入10-7M/L胰島素培養;刺激6 h後採用原位末耑標記法(TUNEL)觀察內皮細胞凋亡、免疫組織化學染色和蛋白印跡法鎗測抗凋亡蛋白bcl-2及eNOS的蛋白錶達變化,數據以錶示,均數比較採用單兇素方差分析,以P<0.05為差異具有統計學意義.結果 與正常對照組比較,燒傷血清促進內皮細胞捌亡(18.5%±3.1%),免疫組化染色bcl-2平均吸光度低(0.14±0.02),蛋白印跡實驗顯示bel-2(0.36 4±0.12)和p-eNOS(0.55±0.28)錶達降低(P<0.01).相對十燒傷血清刺激組,胰島素處理組細胞凋亡顯著減少(9.6 4±2.8%),免疫組化染色bcl-2平均吸光度升高(0.21±0.03),蛋白印跡實驗顯示bcl.2(0.94 4-0.25)和p-eNOS(0.89±0.16)錶達明顯恢複(P<0.01).eNOS在各組中無明顯變化.結論 胰島素明顯抑製燒傷血清誘導下的內皮細胞凋亡併增加擾凋亡蛋白bcl-2的錶達,此作用町能與eNOS的燐痠化有關.
목적 탐토이도소대소상혈청유도혈관내피세포조망적영향급상관궤제.방법 체외상규배양내피세포계ECV304,수궤분위정상대조조(n=6),체적분수15%(V/V)정상대서혈청배양;소상혈청자격조(n=6),체적분수15%(V/V)자제소상대서혈청배양,혈청채자배부30%TBsAIII 탕상대서모형;이도소처리조(n=6),체적분수15%(V/V)자제소상대서혈청중가입10-7M/L이도소배양;자격6 h후채용원위말단표기법(TUNEL)관찰내피세포조망、면역조직화학염색화단백인적법창측항조망단백bcl-2급eNOS적단백표체변화,수거이표시,균수비교채용단흉소방차분석,이P<0.05위차이구유통계학의의.결과 여정상대조조비교,소상혈청촉진내피세포팔망(18.5%±3.1%),면역조화염색bcl-2평균흡광도저(0.14±0.02),단백인적실험현시bel-2(0.36 4±0.12)화p-eNOS(0.55±0.28)표체강저(P<0.01).상대십소상혈청자격조,이도소처리조세포조망현저감소(9.6 4±2.8%),면역조화염색bcl-2평균흡광도승고(0.21±0.03),단백인적실험현시bcl.2(0.94 4-0.25)화p-eNOS(0.89±0.16)표체명현회복(P<0.01).eNOS재각조중무명현변화.결론 이도소명현억제소상혈청유도하적내피세포조망병증가우조망단백bcl-2적표체,차작용정능여eNOS적린산화유관.
Objective To investigate the effeets of insulin on the apoptosis of vascular endothelial cells cuinduced by burn$eruln in order to explore its possible mechamsm.Method Cultured human ECV304 cells were randomly divided into 33x)ups:control group,the ECV304 cells hured by 15%(V/V)rat normal,qertlm(t=6);bum semm group,the ECV3()4 cells simulated by 15%(V/V)self-made burn semm collected from rats with 30%TBSA full-thickness burns on the,back(n=6);and burn Serum+insulin group.the ECV304 cells cultared by insulin(10-7mol/L)and 15%(V/V) seIf-made rat bum serum(n=6).The transferase mediated nick end labeling(TUNEL) method was employed to measure the apoptosis of endothelial cells at 6 hours after stimulation.Meanwhile.immanohistochemical technique and Western blotting were used to determine the protein expressions of bcl-2 and eNOS.Data are expressed ills mean ±SEM.Statistical comparison was made using oneway analysis of vtriance.Significance was accepted at P<0.05.Results Compswith the control group,bum$erunl induced the apoptusis(18.5±3.1%)and down-regalated bcl·2(O.36±0.12)and p-eNOS(O.55±0.28)protein expressions of HUVECs(P<0.01).Burn 9AJ'unl+insulin significantly decreased the apoptosis(9.6 4-2.8%)and up-regulated bcl-2(0.944-0.25)and p-eNOS(0.89±0.16)protein expressions ofHU-VECs in comparison with the bum serllm group(P<0.01).eNOS showed no significant differences in three groups.Conclusions Insulin could markedly inhibit the apoptosis and up-regalate bcl-2 protein expression of HUVECs induced by bum serum,and its mec,harfism might involve the protein expression ofphosphorylated eNOS.
