国际中医中药杂志
國際中醫中藥雜誌
국제중의중약잡지
International Journal of Traditional Chinese Medicine
2015年
9期
803-807
,共5页
九龙藤总黄酮%肌细胞,心肌%过氧化氢%氧化性应激
九龍籐總黃酮%肌細胞,心肌%過氧化氫%氧化性應激
구룡등총황동%기세포,심기%과양화경%양화성응격
BCF%Myocytes%cardiac%Hydrogen peroxide%Oxidative stress
目的 观察九龙藤总黄酮对过氧化氢(H2O2)诱导乳鼠心肌细胞氧化应激损伤的保护作用.方法分离SD大鼠乳鼠心肌细胞,培养72 h后随机分为空白对照组,模型组,九龙藤总黄酮高、中、低剂量组,舒血宁注射液组.除空白对照组外,其余各组细胞经H2O2(100μg/ml)处理.九龙藤总黄酮高、中、低剂量组给予含240、120、60μg/ml九龙藤总黄酮的培养液干预,舒血宁注射液组给予含100μg/ml舒血宁注射液的培养液,空白对照组和模型组给予普通培养基培养.干预6 h后,通过倒置显微镜观察各组细胞形态学变化,通过四甲基偶氮唑蓝(MTT)法检测各组细胞存活率;检测各组细胞培养液中天冬氨酸转氨酶(AST)、磷酸肌酸激酶(CPK)、乳酸脱氢酶(LDH)水平;测定各组细胞中超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、过氧化氢酶(CAT)、丙二醛(MDA)水平;通过流式细胞术测定细胞凋亡率,并通过Western blot方法检测心肌细胞中半胱氨酸天冬氨酸蛋白酶-3(caspase-3)表达.结果与模型组比较,九龙藤总黄酮中、高剂量组心肌细胞培养液中AST[(28.8±6.1)U/ml、(24.5±5.3)U/ml比(36.2± 6.7)U/ml]、CPK[(1.8±0.4)U/ml、(1.5±0.3)U/ml 比(2.5±0.4)U/ml]、LDH[(805.2±160.9)U/L、(671.5± 128.7)U/L比(916.5±168.4)U/L]水平降低(P<0.05或P<0.01);心肌细胞中SOD[(84.8±17.4)U/mg、(95.3±18.2)U/mg比(55.7±13.1)U/mg]、CAT[(23.4±3.1)U/mg、(26.3±3.5)U/mg比(15.2±3.0)U/mg]水平升高(P<0.05 或 P<0.01),MDA[(8.1±1.7)nmol/mg、(6.8±1.5)nmol/mg 比(11.1±2.3)nmol/mg]水平降低(P<0.05或P<0.01),caspase-3[(1.64±0.16)、(1.30±0.12)比(2.06±0.25)]表达、细胞凋亡率[(24.2±5.5)%、(13.4±3.9)%比(51.2±9.1)%]显著降低(P<0.05或P<0.01);九龙藤总黄酮高剂量组细胞中 GSH-Px[(3.6±0.9)U/mg 比(2.4±0.7)U/mg]活性显著升高(P<0.05).结论 九龙藤总黄酮可有效改善 H2O2诱导氧化应激损伤乳鼠心肌细胞形态、提高细胞存活率、改善抗氧化酶活性、下调 caspase-3表达、降低细胞凋亡率,对H2O2诱导乳鼠心肌细胞氧化应激损伤具有剂量依赖性的保护作用.
目的 觀察九龍籐總黃酮對過氧化氫(H2O2)誘導乳鼠心肌細胞氧化應激損傷的保護作用.方法分離SD大鼠乳鼠心肌細胞,培養72 h後隨機分為空白對照組,模型組,九龍籐總黃酮高、中、低劑量組,舒血寧註射液組.除空白對照組外,其餘各組細胞經H2O2(100μg/ml)處理.九龍籐總黃酮高、中、低劑量組給予含240、120、60μg/ml九龍籐總黃酮的培養液榦預,舒血寧註射液組給予含100μg/ml舒血寧註射液的培養液,空白對照組和模型組給予普通培養基培養.榦預6 h後,通過倒置顯微鏡觀察各組細胞形態學變化,通過四甲基偶氮唑藍(MTT)法檢測各組細胞存活率;檢測各組細胞培養液中天鼕氨痠轉氨酶(AST)、燐痠肌痠激酶(CPK)、乳痠脫氫酶(LDH)水平;測定各組細胞中超氧化物歧化酶(SOD)、穀胱甘肽過氧化物酶(GSH-Px)、過氧化氫酶(CAT)、丙二醛(MDA)水平;通過流式細胞術測定細胞凋亡率,併通過Western blot方法檢測心肌細胞中半胱氨痠天鼕氨痠蛋白酶-3(caspase-3)錶達.結果與模型組比較,九龍籐總黃酮中、高劑量組心肌細胞培養液中AST[(28.8±6.1)U/ml、(24.5±5.3)U/ml比(36.2± 6.7)U/ml]、CPK[(1.8±0.4)U/ml、(1.5±0.3)U/ml 比(2.5±0.4)U/ml]、LDH[(805.2±160.9)U/L、(671.5± 128.7)U/L比(916.5±168.4)U/L]水平降低(P<0.05或P<0.01);心肌細胞中SOD[(84.8±17.4)U/mg、(95.3±18.2)U/mg比(55.7±13.1)U/mg]、CAT[(23.4±3.1)U/mg、(26.3±3.5)U/mg比(15.2±3.0)U/mg]水平升高(P<0.05 或 P<0.01),MDA[(8.1±1.7)nmol/mg、(6.8±1.5)nmol/mg 比(11.1±2.3)nmol/mg]水平降低(P<0.05或P<0.01),caspase-3[(1.64±0.16)、(1.30±0.12)比(2.06±0.25)]錶達、細胞凋亡率[(24.2±5.5)%、(13.4±3.9)%比(51.2±9.1)%]顯著降低(P<0.05或P<0.01);九龍籐總黃酮高劑量組細胞中 GSH-Px[(3.6±0.9)U/mg 比(2.4±0.7)U/mg]活性顯著升高(P<0.05).結論 九龍籐總黃酮可有效改善 H2O2誘導氧化應激損傷乳鼠心肌細胞形態、提高細胞存活率、改善抗氧化酶活性、下調 caspase-3錶達、降低細胞凋亡率,對H2O2誘導乳鼠心肌細胞氧化應激損傷具有劑量依賴性的保護作用.
목적 관찰구룡등총황동대과양화경(H2O2)유도유서심기세포양화응격손상적보호작용.방법분리SD대서유서심기세포,배양72 h후수궤분위공백대조조,모형조,구룡등총황동고、중、저제량조,서혈저주사액조.제공백대조조외,기여각조세포경H2O2(100μg/ml)처리.구룡등총황동고、중、저제량조급여함240、120、60μg/ml구룡등총황동적배양액간예,서혈저주사액조급여함100μg/ml서혈저주사액적배양액,공백대조조화모형조급여보통배양기배양.간예6 h후,통과도치현미경관찰각조세포형태학변화,통과사갑기우담서람(MTT)법검측각조세포존활솔;검측각조세포배양액중천동안산전안매(AST)、린산기산격매(CPK)、유산탈경매(LDH)수평;측정각조세포중초양화물기화매(SOD)、곡광감태과양화물매(GSH-Px)、과양화경매(CAT)、병이철(MDA)수평;통과류식세포술측정세포조망솔,병통과Western blot방법검측심기세포중반광안산천동안산단백매-3(caspase-3)표체.결과여모형조비교,구룡등총황동중、고제량조심기세포배양액중AST[(28.8±6.1)U/ml、(24.5±5.3)U/ml비(36.2± 6.7)U/ml]、CPK[(1.8±0.4)U/ml、(1.5±0.3)U/ml 비(2.5±0.4)U/ml]、LDH[(805.2±160.9)U/L、(671.5± 128.7)U/L비(916.5±168.4)U/L]수평강저(P<0.05혹P<0.01);심기세포중SOD[(84.8±17.4)U/mg、(95.3±18.2)U/mg비(55.7±13.1)U/mg]、CAT[(23.4±3.1)U/mg、(26.3±3.5)U/mg비(15.2±3.0)U/mg]수평승고(P<0.05 혹 P<0.01),MDA[(8.1±1.7)nmol/mg、(6.8±1.5)nmol/mg 비(11.1±2.3)nmol/mg]수평강저(P<0.05혹P<0.01),caspase-3[(1.64±0.16)、(1.30±0.12)비(2.06±0.25)]표체、세포조망솔[(24.2±5.5)%、(13.4±3.9)%비(51.2±9.1)%]현저강저(P<0.05혹P<0.01);구룡등총황동고제량조세포중 GSH-Px[(3.6±0.9)U/mg 비(2.4±0.7)U/mg]활성현저승고(P<0.05).결론 구룡등총황동가유효개선 H2O2유도양화응격손상유서심기세포형태、제고세포존활솔、개선항양화매활성、하조 caspase-3표체、강저세포조망솔,대H2O2유도유서심기세포양화응격손상구유제량의뢰성적보호작용.
Objective To investigate the protective effects of Bauhinia championii Benth flavones(BCF) on oxidative stress of neonatal rat cadiocytes induced by H2O2.Methods Cadiocytes of neonate rat was cultivated for 72 hours and divided into six groups: a normal control group, a H2O2 group, BCF(60, 120 and 240μg/ml)+H2O2 groups and aShuxueninginjection (100μg/ml)+H2O2 group (n=8). 6 hours after the drugs were given, the morphology changes was observed and the survival rate was detected; the content of AST, CPK, LDH in culture medium were detected; the activity of SOD, CAT, GSH-Px and the content of MDA in cardiomyocytes were also determinted; the apoptosis rate were detected, and the expression of caspase-3 in cardiomyocytes was detected by Western blot.Results Compared with the H2O2 group, the activity of AST(28.8 ± 6.1 U/ml, 24.5 ± 5.3 U/mlvs. 36.2 ± 6.7 U/ml), CPK(1.8 ± 0.4 U/ml, 1.5 ± 0.3 U/mlvs. 2.5 ±0.4 U/ml), LDH(805.2 ± 160.9 U/L, 671.5 ± 128.7 U/Lvs. 916.5 ± 168.4 U/L) in culture medium were significantly decreased (P<0.05,P<0.01), the activity of SOD(84.8 ± 17.4 U/mg, 95.3 ± 18.2 U/mgvs. 55.7 ± 13.1 U/mg), CAT(23.4 ± 3.1 U/mg, 26.3 ± 3.5U/mgvs. 15.2 ± 3.0 U/mg) in cardiomyocytes were significantly increased and the content of MDA(8.1 ± 1.7 nmol/mg, 6.8 ± 1.5 nmol/mgvs. 11.1 ± 2.3 nmol/mg) were significantly decreased (P<0.05,P<0.01), the expression of c caspase-3(1.64 ± 0.16, 1.30 ± 0.12vs. 2.06 ± 0.25) and the apoptosis rate (24.2% ± 5.5%, 13.4% ± 3.9%vs. 51.2% ± 9.1%) were significantly decreased (P<0.05,P<0.01); the activity of GSH-Px (3.6 ± 0.9 U/mg vs. 2.4 ± 0.7 U/mg) in cardiomyocytes of BCF 240μg/ml treatment group was increased (P<0.05).Conclusions BCF could effectively improve the morphology of neonatal rat cadiocytes induced by H2O2, increase the survival rate, improve the activity of antioxidase, down-regulate the expression of caspase-3 and decrease the apoptosis rate, suggesting that BCF had dose-dependent protective effects on oxidative stress of neonatal rat cadiocytes induced by H2O2.
