中国生化药物杂志
中國生化藥物雜誌
중국생화약물잡지
CHINESE JOURNAL OF BIOCHEMICAL PHARMACEUTICS
2014年
7期
27-30
,共4页
徐庆国%谈金强%宋希林%陈沅
徐慶國%談金彊%宋希林%陳沅
서경국%담금강%송희림%진원
葛花总黄酮%阿霉素%脂质过氧化%细胞凋亡%Bax%Bcl-2
葛花總黃酮%阿黴素%脂質過氧化%細胞凋亡%Bax%Bcl-2
갈화총황동%아매소%지질과양화%세포조망%Bax%Bcl-2
flos puerariae flavonoid%adriamycin%myocardial lipid peroxidation%apoptosis%Bax%Bcl-2
目的:探讨葛花总黄酮对阿霉素中毒性心肌炎的保护作用及其机制。方法健康昆明种小鼠96只,雄雌不拘,分为6组,每组16只。正常对照组(control):每天经口灌胃(ig)生理盐水(NS)20 mL/kg,并于实验第2天起,腹腔注射(ip)NS 10 mL/kg,隔天1次,共7次;阿霉素模型对照组(model):每天ig 20 mL/kg NS,同时于实验第2天起,ip阿霉素3 mg/kg,隔天1次,共7次;阿霉素+葛花总黄酮小剂量组(50 mg/kg);阿霉素+葛花总黄酮中剂量组(100 mg/kg);阿霉素+葛花总黄酮大剂量组(200 mg/kg);维生素E阳性对照组(Vit E):造模同阿霉素模型对照组,同时每天ig Vit E 0.04 g/kg。阿霉素+葛花总黄酮各组的造模方法同阿霉素模型对照组,同时每天分别ig葛花总黄酮相应剂量,各组实验连续进行15 d。采用HE染色检测心肌组织结构变化,采用免疫组织法检测心肌组织凋亡相关蛋白Bax和Bcl-2表达情况,并进行组织学检查,观察心肌结构变化。结果与正常对照组比较,阿霉素(3 mg/kg,ip,7次)可致小鼠血清肌酸激酶(CK)、谷草转氨酶(GOT)、乳酸脱氢酶(LDH)和iNOS 活力均显著升高(P<0.01),同时使心肌丙二醛(MDA)含量明显上升(P<0.01),而超氧化物歧化酶(SOD)活力则显著下降(P<0.01)。另外,小鼠心肌细胞凋亡数明显升高,细胞凋亡率达(40.5±5.2)%,凋亡相关蛋白Bax和Bcl-2含量均明显升高,与正常对照组比较差异有显著性(P<0.01),但Bcl-2/Bax值降低。葛花总黄酮(50、100、200 mg/kg,15 d)和Vit E组剂量依赖性的逆转阿霉素所致的上述改变,表现为降低小鼠血清CK、GOT、LDH和iNOS活力,降低心肌MDA含量和增加SOD活力,降低促凋亡基因Bax蛋白表达,使Bcl-2/Bax值增加,尤其以高剂量组作用最为明显。光镜和电镜结果对心肌形态学的观察也证实了葛花总黄酮对阿霉素中毒性心肌炎保护作用。结论阿霉素可以诱导心肌细胞凋亡,引起实验小鼠中毒性心肌炎。葛花总黄酮能拮抗阿霉素所致的心肌损伤,其作用机制与增强心肌SOD活力和抗心肌脂质过氧化、降低凋亡基因Bax蛋白表达等有关。
目的:探討葛花總黃酮對阿黴素中毒性心肌炎的保護作用及其機製。方法健康昆明種小鼠96隻,雄雌不拘,分為6組,每組16隻。正常對照組(control):每天經口灌胃(ig)生理鹽水(NS)20 mL/kg,併于實驗第2天起,腹腔註射(ip)NS 10 mL/kg,隔天1次,共7次;阿黴素模型對照組(model):每天ig 20 mL/kg NS,同時于實驗第2天起,ip阿黴素3 mg/kg,隔天1次,共7次;阿黴素+葛花總黃酮小劑量組(50 mg/kg);阿黴素+葛花總黃酮中劑量組(100 mg/kg);阿黴素+葛花總黃酮大劑量組(200 mg/kg);維生素E暘性對照組(Vit E):造模同阿黴素模型對照組,同時每天ig Vit E 0.04 g/kg。阿黴素+葛花總黃酮各組的造模方法同阿黴素模型對照組,同時每天分彆ig葛花總黃酮相應劑量,各組實驗連續進行15 d。採用HE染色檢測心肌組織結構變化,採用免疫組織法檢測心肌組織凋亡相關蛋白Bax和Bcl-2錶達情況,併進行組織學檢查,觀察心肌結構變化。結果與正常對照組比較,阿黴素(3 mg/kg,ip,7次)可緻小鼠血清肌痠激酶(CK)、穀草轉氨酶(GOT)、乳痠脫氫酶(LDH)和iNOS 活力均顯著升高(P<0.01),同時使心肌丙二醛(MDA)含量明顯上升(P<0.01),而超氧化物歧化酶(SOD)活力則顯著下降(P<0.01)。另外,小鼠心肌細胞凋亡數明顯升高,細胞凋亡率達(40.5±5.2)%,凋亡相關蛋白Bax和Bcl-2含量均明顯升高,與正常對照組比較差異有顯著性(P<0.01),但Bcl-2/Bax值降低。葛花總黃酮(50、100、200 mg/kg,15 d)和Vit E組劑量依賴性的逆轉阿黴素所緻的上述改變,錶現為降低小鼠血清CK、GOT、LDH和iNOS活力,降低心肌MDA含量和增加SOD活力,降低促凋亡基因Bax蛋白錶達,使Bcl-2/Bax值增加,尤其以高劑量組作用最為明顯。光鏡和電鏡結果對心肌形態學的觀察也證實瞭葛花總黃酮對阿黴素中毒性心肌炎保護作用。結論阿黴素可以誘導心肌細胞凋亡,引起實驗小鼠中毒性心肌炎。葛花總黃酮能拮抗阿黴素所緻的心肌損傷,其作用機製與增彊心肌SOD活力和抗心肌脂質過氧化、降低凋亡基因Bax蛋白錶達等有關。
목적:탐토갈화총황동대아매소중독성심기염적보호작용급기궤제。방법건강곤명충소서96지,웅자불구,분위6조,매조16지。정상대조조(control):매천경구관위(ig)생리염수(NS)20 mL/kg,병우실험제2천기,복강주사(ip)NS 10 mL/kg,격천1차,공7차;아매소모형대조조(model):매천ig 20 mL/kg NS,동시우실험제2천기,ip아매소3 mg/kg,격천1차,공7차;아매소+갈화총황동소제량조(50 mg/kg);아매소+갈화총황동중제량조(100 mg/kg);아매소+갈화총황동대제량조(200 mg/kg);유생소E양성대조조(Vit E):조모동아매소모형대조조,동시매천ig Vit E 0.04 g/kg。아매소+갈화총황동각조적조모방법동아매소모형대조조,동시매천분별ig갈화총황동상응제량,각조실험련속진행15 d。채용HE염색검측심기조직결구변화,채용면역조직법검측심기조직조망상관단백Bax화Bcl-2표체정황,병진행조직학검사,관찰심기결구변화。결과여정상대조조비교,아매소(3 mg/kg,ip,7차)가치소서혈청기산격매(CK)、곡초전안매(GOT)、유산탈경매(LDH)화iNOS 활력균현저승고(P<0.01),동시사심기병이철(MDA)함량명현상승(P<0.01),이초양화물기화매(SOD)활력칙현저하강(P<0.01)。령외,소서심기세포조망수명현승고,세포조망솔체(40.5±5.2)%,조망상관단백Bax화Bcl-2함량균명현승고,여정상대조조비교차이유현저성(P<0.01),단Bcl-2/Bax치강저。갈화총황동(50、100、200 mg/kg,15 d)화Vit E조제량의뢰성적역전아매소소치적상술개변,표현위강저소서혈청CK、GOT、LDH화iNOS활력,강저심기MDA함량화증가SOD활력,강저촉조망기인Bax단백표체,사Bcl-2/Bax치증가,우기이고제량조작용최위명현。광경화전경결과대심기형태학적관찰야증실료갈화총황동대아매소중독성심기염보호작용。결론아매소가이유도심기세포조망,인기실험소서중독성심기염。갈화총황동능길항아매소소치적심기손상,기작용궤제여증강심기SOD활력화항심기지질과양화、강저조망기인Bax단백표체등유관。
Objective To investigate the protective effect of flos puerariae flavonoid on adriamycin (ADR)-induced toxic myocarditis and its mechanisms from morphological,biochemical and molecular levels.Methods 96 healthy Kunming male mice were randomly divided into 6 groups:normal control group,ADR model control group,ADR+low dose of flos puerariae flavonoid group(50 mg/kg),ADR +middle dose of flos puerariae flavonoid group(100 mg/kg),ADR +high dose of flos puerariae flavonoid group(200 mg/kg),and Vit E positive control group(40 mg/kg),16 in each group.The drugs were orally administered for consecutive 15 d and the model of toxic myocarditis was induced by intraperitoneal injection of ADR(3 mg/kg)in mice from day 2,one time every other day,for 7 times Colorimetry was used to measure the changes of marker enzymes about myocardial injury and inducible nitric oxide synthase(iNOS)activity in serum and tissue;immunohistochemical method was adopted to detecte the expression of myocardial apoptosis related proteins Bax and bcl-2;HE staining was conducted to observe the pathological changes of cardiac structure.Results Compared with normal control group,ADR(3 mg/kg,ip,7 times)induced the elevation of serum creatine kinase (CK),lactate dehydrogenase (LDH),aspartate transaminase(GOT)and iNOS activity increased significantly in mice(P<0.01).Meanwhile myocardial superoxide dismutase(SOD)activity decreased, and the malondialdehyde(MDA)content increased(P<0.01).Myocardial cell apoptosis in mice increased significantly,and the apoptosis rate was(40.5 ± 5.2)%;the expressions of Bax and Bcl-2 were significantly increased(P<0.01),However,the Bcl-2/Bax ratio decreased.The flos puerariae flavonoid (50,100,200 mg/kg,ig,15 d)and Vit E positive control group could reverse the changes induced by ADR,decrease serum CK,LDH,GOT and iNOS activities,increased myocardial SOD activity,lower MDA content and the expression of bax protein,and elevated Bcl-2/Bax ratio,in a dose-dependent manner.Light microscopy confirmed that flos puerariae flavonoid significantly alleviated the changes of myocardial microstructure.Conclusion ADR could induce myocardial cell apoptosis and lead toxic myocarditis in experimental mice.The flos puerariae flavonoid has protective effect on ADR-induced myocardial injury and the mechanism may be related to elevating myocardial SOD activity and anti-lipid peroxidation,inhibiting the expression of Bax protein and adriamycin-induced cardiomyocyte apoptosis.