世界科学技术-中医药现代化
世界科學技術-中醫藥現代化
세계과학기술-중의약현대화
WORLD SCIENCE AND TECHNOLOGY-MODERNIZATION OF TRADITIONAL CHINESE MEDICINE
2013年
8期
1725-1734
,共10页
CORM-2%神经细胞凋亡%Caspase%线粒体途径%醒脑静注射液
CORM-2%神經細胞凋亡%Caspase%線粒體途徑%醒腦靜註射液
CORM-2%신경세포조망%Caspase%선립체도경%성뇌정주사액
CORM-2%neuronal apoptosis%Caspase%mitochondrial pathway%Xingnaojing Injection
目的：探讨外源性一氧化碳释放剂（CORM-2）诱导大鼠神经细胞凋亡及醒脑静注射液对其干预作用的机制。方法：在光学显微镜下观察CORM-2对神经细胞形态的影响；通过MTT法检测CORM-2对细胞存活率的影响；应用流式细胞仪检测细胞凋亡率；以Western Blotting检测相关蛋白的表达。同时观察醒脑静注射液对神经细胞形态，细胞存活率及相关蛋白表达的影响。结果：CORM-2影响神经细胞存活率呈时间依赖性和剂量依赖性；100、200、400、800μmol·L-1 CORM-2作用于神经细胞24 h后，可使细胞存活率逐渐降低；可以诱导神经细胞凋亡及Caspase依赖性线粒体途经相关蛋白Caspase-3、Caspase-9及Cytochrome C（Cyt C）的活化，且呈剂量依赖性。选择200μmol·L-1 CORM-2对皮层神经细胞进行诱导损伤，经醒脑静注射液孵育神经细胞20 h后，10 mL·L-1及20 mL·L-1醒脑静注射液可明显降低细胞早期凋亡率，细胞Caspase-3、Caspase-9与Cyt C的表达逐渐降低。结论：CORM-2可能通过激活Caspase依赖性线粒体途径诱导神经细胞凋亡，醒脑静注射液可抑制细胞凋亡的线粒体途径从而干预CORM-2诱导的神经细胞凋亡。
目的：探討外源性一氧化碳釋放劑（CORM-2）誘導大鼠神經細胞凋亡及醒腦靜註射液對其榦預作用的機製。方法：在光學顯微鏡下觀察CORM-2對神經細胞形態的影響；通過MTT法檢測CORM-2對細胞存活率的影響；應用流式細胞儀檢測細胞凋亡率；以Western Blotting檢測相關蛋白的錶達。同時觀察醒腦靜註射液對神經細胞形態，細胞存活率及相關蛋白錶達的影響。結果：CORM-2影響神經細胞存活率呈時間依賴性和劑量依賴性；100、200、400、800μmol·L-1 CORM-2作用于神經細胞24 h後，可使細胞存活率逐漸降低；可以誘導神經細胞凋亡及Caspase依賴性線粒體途經相關蛋白Caspase-3、Caspase-9及Cytochrome C（Cyt C）的活化，且呈劑量依賴性。選擇200μmol·L-1 CORM-2對皮層神經細胞進行誘導損傷，經醒腦靜註射液孵育神經細胞20 h後，10 mL·L-1及20 mL·L-1醒腦靜註射液可明顯降低細胞早期凋亡率，細胞Caspase-3、Caspase-9與Cyt C的錶達逐漸降低。結論：CORM-2可能通過激活Caspase依賴性線粒體途徑誘導神經細胞凋亡，醒腦靜註射液可抑製細胞凋亡的線粒體途徑從而榦預CORM-2誘導的神經細胞凋亡。
목적：탐토외원성일양화탄석방제（CORM-2）유도대서신경세포조망급성뇌정주사액대기간예작용적궤제。방법：재광학현미경하관찰CORM-2대신경세포형태적영향；통과MTT법검측CORM-2대세포존활솔적영향；응용류식세포의검측세포조망솔；이Western Blotting검측상관단백적표체。동시관찰성뇌정주사액대신경세포형태，세포존활솔급상관단백표체적영향。결과：CORM-2영향신경세포존활솔정시간의뢰성화제량의뢰성；100、200、400、800μmol·L-1 CORM-2작용우신경세포24 h후，가사세포존활솔축점강저；가이유도신경세포조망급Caspase의뢰성선립체도경상관단백Caspase-3、Caspase-9급Cytochrome C（Cyt C）적활화，차정제량의뢰성。선택200μmol·L-1 CORM-2대피층신경세포진행유도손상，경성뇌정주사액부육신경세포20 h후，10 mL·L-1급20 mL·L-1성뇌정주사액가명현강저세포조기조망솔，세포Caspase-3、Caspase-9여Cyt C적표체축점강저。결론：CORM-2가능통과격활Caspase의뢰성선립체도경유도신경세포조망，성뇌정주사액가억제세포조망적선립체도경종이간예CORM-2유도적신경세포조망。
This study was aimed to investigate the mechanism of CO-releasing molecules (CORM-2) induced apoptosis and the intervention effect of Xingnaojing (XNJ) Injection in neuronal cells of rats. Optical microscope was applied to observe morphologic changes of neuronal cells. MTT assay was performed to assess the survival rates of CORM-2 on neuronal cells. Apoptosis was examined by flow cytometric analysis and the expression of relative proteins was measured by western blotting analysis. At the same time the morphologic changes, survival rates and expression of relative proteins of neuronal cells were also checked after XNJ treatment. The results showed that CORM-2 can influence survival rates in a time- and concentration-dependent manner. Survival rates decreased gradually after the cultures subjected to 24 h with 100 μmol·L-1, 200 μmol·L-1, 400 μmol·L-1 and 800 μmol·L-1 of CORM-2. It can induce neuronal apoptosis and activate Caspase-3, Caspase-9 and Cytochrome C in a concentration-dependent manner. The neuronal cells were treated with 200 μmol·L-1 of CORM-2 and then incubated with 10 mL·L-1 and 20 mL·L-1 XNJ injection for 20 h. It turned out early neuronal apoptosis decreased and the expression of Caspase-3, Caspase-9 and Cytochrome C also decreased. To sum up, CORM-2 may induce neuronal apoptosis through Caspase-dependent mitochondrial pathway, which can be intervened by XNJ Injection through inhibiting Caspase-dependent mitochondrial pathway.
