国际麻醉学与复苏杂志
國際痳醉學與複囌雜誌
국제마취학여복소잡지
INTERNATIONAL JOURNAL OF ANESTHESIOLOGY AND RESUSCITATION
2015年
5期
428-431
,共4页
脂多糖%脐静脉内皮细胞%凋亡%右美托咪定
脂多糖%臍靜脈內皮細胞%凋亡%右美託咪定
지다당%제정맥내피세포%조망%우미탁미정
Lipopolysaccharide%Human umbilical vein endothelial cells%Apoptosis%Dexmedetomidine
目的 评价右美托咪定对脂多糖(lipopolysaccharide,LPS)诱导的血管内皮细胞凋亡的影响.方法 参照随机数字表法将人脐静脉内皮细胞HUVEC-12随机分为4组(每组20孔):正常对照组(C组)、右美托咪定组(D组)、LPS组(L 组)、LPS+右美托咪定组(L+D组),培养24 h后:四甲基偶氮唑盐微量酶反应比色法(MTT法)和流式细胞术分别检测细胞活力和细胞凋亡率,黄嘌呤氧化酶法和硫代巴比妥酸法(thiobarbituric acid,TBA)测定各组细胞超氧化物歧化酶(superoxidedismutase,SOD)的活性和丙二醛(malonaldehyde,MDA)的含量,Western blot法检测细胞多聚腺苷酸二磷酸-1(Poly-ADP Ribosy polymerase-1,PARP-1)蛋白裂解片段的表达.结果 L组和L+D组细胞活力分别是0.95±0.08和1.08±0.10(P<0.05),与C组比较,分别降低36%和27%;L组和L+D组细胞凋亡率分别是(14.7±1.8)%和(8.8±1.1)%(P<0.05),分别是C组的2.6倍和1.1倍;L组和L+D组细胞SOD活性分别是(99±6) U/mg和(182±9) U/mg(P<0.05),与C组比较,分别降低53%和14%;L组和L+D组细胞MDA含量分别是(29.9±1.8) nmol/mg和(19.3±2.1) nmol/mg(P<0.05),分别是C组的1.6倍和79%;L组和L+D组细胞内PARP-1片段(相对分子质量89×103)表达分别是1.152±0.095和0.564±0.045 (P<0.05),分别是C组的4.8倍和1.8倍;D组上述指标的差异无统计学意义(P>0.05).结论 右美托咪定可有效减少LPS诱导下脐静脉内皮细胞的凋亡,其机制可能与抑制氧化应激、下调PARP-1裂解片段的表达有关.
目的 評價右美託咪定對脂多糖(lipopolysaccharide,LPS)誘導的血管內皮細胞凋亡的影響.方法 參照隨機數字錶法將人臍靜脈內皮細胞HUVEC-12隨機分為4組(每組20孔):正常對照組(C組)、右美託咪定組(D組)、LPS組(L 組)、LPS+右美託咪定組(L+D組),培養24 h後:四甲基偶氮唑鹽微量酶反應比色法(MTT法)和流式細胞術分彆檢測細胞活力和細胞凋亡率,黃嘌呤氧化酶法和硫代巴比妥痠法(thiobarbituric acid,TBA)測定各組細胞超氧化物歧化酶(superoxidedismutase,SOD)的活性和丙二醛(malonaldehyde,MDA)的含量,Western blot法檢測細胞多聚腺苷痠二燐痠-1(Poly-ADP Ribosy polymerase-1,PARP-1)蛋白裂解片段的錶達.結果 L組和L+D組細胞活力分彆是0.95±0.08和1.08±0.10(P<0.05),與C組比較,分彆降低36%和27%;L組和L+D組細胞凋亡率分彆是(14.7±1.8)%和(8.8±1.1)%(P<0.05),分彆是C組的2.6倍和1.1倍;L組和L+D組細胞SOD活性分彆是(99±6) U/mg和(182±9) U/mg(P<0.05),與C組比較,分彆降低53%和14%;L組和L+D組細胞MDA含量分彆是(29.9±1.8) nmol/mg和(19.3±2.1) nmol/mg(P<0.05),分彆是C組的1.6倍和79%;L組和L+D組細胞內PARP-1片段(相對分子質量89×103)錶達分彆是1.152±0.095和0.564±0.045 (P<0.05),分彆是C組的4.8倍和1.8倍;D組上述指標的差異無統計學意義(P>0.05).結論 右美託咪定可有效減少LPS誘導下臍靜脈內皮細胞的凋亡,其機製可能與抑製氧化應激、下調PARP-1裂解片段的錶達有關.
목적 평개우미탁미정대지다당(lipopolysaccharide,LPS)유도적혈관내피세포조망적영향.방법 삼조수궤수자표법장인제정맥내피세포HUVEC-12수궤분위4조(매조20공):정상대조조(C조)、우미탁미정조(D조)、LPS조(L 조)、LPS+우미탁미정조(L+D조),배양24 h후:사갑기우담서염미량매반응비색법(MTT법)화류식세포술분별검측세포활력화세포조망솔,황표령양화매법화류대파비타산법(thiobarbituric acid,TBA)측정각조세포초양화물기화매(superoxidedismutase,SOD)적활성화병이철(malonaldehyde,MDA)적함량,Western blot법검측세포다취선감산이린산-1(Poly-ADP Ribosy polymerase-1,PARP-1)단백렬해편단적표체.결과 L조화L+D조세포활력분별시0.95±0.08화1.08±0.10(P<0.05),여C조비교,분별강저36%화27%;L조화L+D조세포조망솔분별시(14.7±1.8)%화(8.8±1.1)%(P<0.05),분별시C조적2.6배화1.1배;L조화L+D조세포SOD활성분별시(99±6) U/mg화(182±9) U/mg(P<0.05),여C조비교,분별강저53%화14%;L조화L+D조세포MDA함량분별시(29.9±1.8) nmol/mg화(19.3±2.1) nmol/mg(P<0.05),분별시C조적1.6배화79%;L조화L+D조세포내PARP-1편단(상대분자질량89×103)표체분별시1.152±0.095화0.564±0.045 (P<0.05),분별시C조적4.8배화1.8배;D조상술지표적차이무통계학의의(P>0.05).결론 우미탁미정가유효감소LPS유도하제정맥내피세포적조망,기궤제가능여억제양화응격、하조PARP-1렬해편단적표체유관.
Objective To study the effect of dexmedetomidine on lipopolysaccharide (LPS)-induced apoptosis in human umbilical vein endothelial cells.Methods Human umbilical vein endothelial cells were randomly divided into four group (n=20):normal control group (group C),dexmedetomidine group (group D),LPS group (group L),LPS plus dexmedetomidine group(group L+D).The cell viability and apoptosis was measured by MTT assay and flow cytometry respectively after 24 h of cell culture.The superoxide dismutase (SOD) activity and malonaldehyde(MDA) content was measured by xanthine oxidase method and thiobarbituric acid (TBA) test respectively.The expression of cleaved Poly-ADP Ribosy polymerase-1 (PARP-1) protein was detected by Western blot.Results The cell survival rate in group L and group L+D were 0.95±0.08 and 1.08±0.10(P<0.05),which were decreased by 36% and 27% respectively when compared with group C.The apoptotic rate in group L and group L+D were(14.70±1.8)% and (8.80±1.1)% (P<0.05),which were increased by 2.6 times and 1.1 times respectively when compared with group C.SOD activity in group L and group L+D were (99±6) U/mg and (182±9) U/mg (P<0.05),which were decreased by 53% and 14% respectively when compared with group C.MDA content in group L and group L+D were (29.9±1.8) nmol/mg and (19.3±2.1) nmol/mg(P<0.05),which were increased by 1.6 times and 79% respectively when compared with group C.The expression of PARP-1 protein fragment in group L and group L+D were 1.152±0.095 and 0.564±0.045 (P<0.05),which were increased by 4.8 times and 1.8 times respectively when compared with group C.No significant difference was found in group D when compared with group C (P>0.05).Conclusions Dexmedetomidine can effectively reduce LPS-induced apoptosis in human umbilical vein endothelial cells by inhibiting oxidative stress and down-regulating expression of PARP-1 protein fragment.