中国药理学通报
中國藥理學通報
중국약이학통보
Chinese Pharmacological Bulletin
2015年
11期
1510-1515
,共6页
张晓晖%曾繁典%孙智达%杨卓欣%熊益群%徐超英%刘心亮%林坚%穆桂萍%徐绍钢%刘文赫
張曉暉%曾繁典%孫智達%楊卓訢%熊益群%徐超英%劉心亮%林堅%穆桂萍%徐紹鋼%劉文赫
장효휘%증번전%손지체%양탁흔%웅익군%서초영%류심량%림견%목계평%서소강%류문혁
原花青素B2%脂多糖%心肌细胞%凋亡%NADPH氧化酶%活性氧自由基
原花青素B2%脂多糖%心肌細胞%凋亡%NADPH氧化酶%活性氧自由基
원화청소B2%지다당%심기세포%조망%NADPH양화매%활성양자유기
procyanidin B2%lipopolysacchride%car-diomyocyte%apoptosis%NADPH oxidase%reactive oxy-gen species
目的：研究原花青素B2( procyanidin B2, PCB2)对脂多糖( lipopolysacchride, LPS)诱导的心肌细胞凋亡的保护作用及其机制。方法采用原代培养新生大鼠心肌细胞, LPS诱导心肌细胞损伤模型,PCB2低、中、高剂量组分别用含有6．25、12．5、25．0μmol·L-1 PCB2的DMEM培养基持续培养24 h。采用四唑盐( MTT)比色法测定心肌细胞存活率,光泽精化学发光法测定心肌细胞NOX的活性, Western blot法分析心肌NADPH氧化酶p47phox亚基的表达,TUNEL法检测细胞凋亡,流式细胞术测定心肌细胞ROS的含量。结果 LPS诱导的细胞损伤组与正常对照组( Control)比较,心肌细胞活力明显降低( P<0．01),而心肌细胞NOX活性、p47 phox亚基的表达、凋亡细胞数量以及 ROS 含量均明显增加( P <0．01)。 PCB2处理后,低、中、高剂量组细胞活力均明显升高,心肌细胞NOX活性、p47 phox亚基的表达、凋亡细胞数量以及ROS含量均明显下降,且具有剂量依赖性( P<0．01)。结论 PCB2通过抑制NADPH氧化酶激活、p47 phox的表达以及活性氧的产生发挥对LPS诱导的心肌细胞凋亡的保护作用。
目的：研究原花青素B2( procyanidin B2, PCB2)對脂多糖( lipopolysacchride, LPS)誘導的心肌細胞凋亡的保護作用及其機製。方法採用原代培養新生大鼠心肌細胞, LPS誘導心肌細胞損傷模型,PCB2低、中、高劑量組分彆用含有6．25、12．5、25．0μmol·L-1 PCB2的DMEM培養基持續培養24 h。採用四唑鹽( MTT)比色法測定心肌細胞存活率,光澤精化學髮光法測定心肌細胞NOX的活性, Western blot法分析心肌NADPH氧化酶p47phox亞基的錶達,TUNEL法檢測細胞凋亡,流式細胞術測定心肌細胞ROS的含量。結果 LPS誘導的細胞損傷組與正常對照組( Control)比較,心肌細胞活力明顯降低( P<0．01),而心肌細胞NOX活性、p47 phox亞基的錶達、凋亡細胞數量以及 ROS 含量均明顯增加( P <0．01)。 PCB2處理後,低、中、高劑量組細胞活力均明顯升高,心肌細胞NOX活性、p47 phox亞基的錶達、凋亡細胞數量以及ROS含量均明顯下降,且具有劑量依賴性( P<0．01)。結論 PCB2通過抑製NADPH氧化酶激活、p47 phox的錶達以及活性氧的產生髮揮對LPS誘導的心肌細胞凋亡的保護作用。
목적：연구원화청소B2( procyanidin B2, PCB2)대지다당( lipopolysacchride, LPS)유도적심기세포조망적보호작용급기궤제。방법채용원대배양신생대서심기세포, LPS유도심기세포손상모형,PCB2저、중、고제량조분별용함유6．25、12．5、25．0μmol·L-1 PCB2적DMEM배양기지속배양24 h。채용사서염( MTT)비색법측정심기세포존활솔,광택정화학발광법측정심기세포NOX적활성, Western blot법분석심기NADPH양화매p47phox아기적표체,TUNEL법검측세포조망,류식세포술측정심기세포ROS적함량。결과 LPS유도적세포손상조여정상대조조( Control)비교,심기세포활력명현강저( P<0．01),이심기세포NOX활성、p47 phox아기적표체、조망세포수량이급 ROS 함량균명현증가( P <0．01)。 PCB2처리후,저、중、고제량조세포활력균명현승고,심기세포NOX활성、p47 phox아기적표체、조망세포수량이급ROS함량균명현하강,차구유제량의뢰성( P<0．01)。결론 PCB2통과억제NADPH양화매격활、p47 phox적표체이급활성양적산생발휘대LPS유도적심기세포조망적보호작용。
Aim To study the mechanisms of the pro-tective effect of procyanidin B2 ( PCB2 ) on the myocar-dial cell apoptosis induced by lipopolysaccharide ( LPS) . Methods Using the primary culture rat myo-cardial cells, myocardial cell injury model was induced by LPS. PCB2 low, medium and high dose groups, were cultured with 6. 25 , 12. 5 , 25. 0 μmol · L-1 PCB2 respectively in DMEM medium for 24 h continu-ously. Myocardial cell survival rate was determined by MTT colorimetric method. Cardiacmyocyte NOX activi-ty was determined by lucigen chemiluminescence meth-od . Western blot analysis was used to detect myocardi-al NADPH oxidase p47phox expression. TUNEL method was used to detect apoptosis and flow cytometry was used to determine the content of myocardial cells ROS. Results Compared with control group, the cell dam-age induced by LPS group myocardial cell survival rate significantly decreased ( P <0. 01 ) , and myocardial cell NOX activity, p47phox expression, apoptotic cell number and ROS content were significantly increased (P<0. 01). PCB2 low, medium and high dose groups cell survival rates were significantly elevated, myocar-dial cell NOX activity and p47phox expression, apoptotic cell number and the ROS content decreased significant-ly in a dose-dependent manner ( P <0. 01 ) . Conclu-sion PCB2 protects myocardial cell apoptosis induced by LPS via inhibiting the expression of NADPH oxidase activation, p47phox expression and reactive oxygen spe-cies generation.
