出 处:《Neural Regeneration Research》2007年第6期369-371,共3页中国神经再生研究(英文版)
基 金:Initial Funding for Doctor from Guangdong Science and Technology Bureau, No. 06300709;Initial Funding for Doctor from Guangdong Pharmaceutical College, No. 43543096
摘 要:BACKGROUND: Extracts of ginkgo biloba leaves (EGB) and its metabolites have been reported to enhance brain function and nerve behavior. It has also been hypothesized that they can protect neurons from oxidative stress. OBJECTIVE: To investigate protective effects of EGB on peroxide (H2O2)-induced oxidative stress damage in PC12 cells. DESIGN: Observational contrast study. SETTING: Department ofPathophysiology, Guangdong Pharmacological College. MATERIALS: EGB was provided by Xi'an Fujie Biotechnological Development Company; 1640 culture medium, methylthiazolyl tetrazolium (MTT), trypsin and dimathyl sulfoxide (DMSO) by Sigma Company; PC12 cell strain by Cell Center of Medical College of Zhongshan University; calf serum by Hangzhou Sijiqing Bioengineering Company; lactate dehydrogenase (LDH) kit by Nanjing Jiancheng Bioengineering Research Institute. METHODS: The experiment was carried out in Department of Cell Biology of Guangdong Pharmacological College from June to December 2005. ①Cell culture: PC12 cells were cultured in 1640 medium containing 200 g/L fetal calf serum. The cells were diluted to 1 × 10^7 L^-1 and washed every two days. Those cells were used to experiment until they grew in logarithm on solid wall. ② Grouping and intervention: PC12 cells (1 × 10^8L^-1) were plated in 96-well plates with the density of 200 μ L/hole and divided into three groups: normal control group (routinely adding media), H2O2 group (treating with media and H2O2 for 20 hours) and EGB group (adding media, 100μmol/L EGB and 100 μmol/L H2O2). ③ MTT assay: PC12 cells (1 × 10^8L^-1) were plated in 96-well plates and divided into three groups with 8 holes for each group. Under sterile condition, cells were added with 5 g/L MTT (100μL) and cultured for 4 hours. And then, 200 μ L DMSO fluid was added and shaken for 30 minutes until blue crystal products formed were dissolved soundly ④ Experimental evaluation: Absorbance (A) at 630 nm was measured aBACKGROUND: Extracts of ginkgo biloba leaves (EGB) and its metabolites have been reported to enhance brain function and nerve behavior. It has also been hypothesized that they can protect neurons from oxidative stress. OBJECTIVE: To investigate protective effects of EGB on peroxide (H2O2)-induced oxidative stress damage in PC12 cells. DESIGN: Observational contrast study. SETTING: Department ofPathophysiology, Guangdong Pharmacological College. MATERIALS: EGB was provided by Xi'an Fujie Biotechnological Development Company; 1640 culture medium, methylthiazolyl tetrazolium (MTT), trypsin and dimathyl sulfoxide (DMSO) by Sigma Company; PC12 cell strain by Cell Center of Medical College of Zhongshan University; calf serum by Hangzhou Sijiqing Bioengineering Company; lactate dehydrogenase (LDH) kit by Nanjing Jiancheng Bioengineering Research Institute. METHODS: The experiment was carried out in Department of Cell Biology of Guangdong Pharmacological College from June to December 2005. ①Cell culture: PC12 cells were cultured in 1640 medium containing 200 g/L fetal calf serum. The cells were diluted to 1 × 10^7 L^-1 and washed every two days. Those cells were used to experiment until they grew in logarithm on solid wall. ② Grouping and intervention: PC12 cells (1 × 10^8L^-1) were plated in 96-well plates with the density of 200 μ L/hole and divided into three groups: normal control group (routinely adding media), H2O2 group (treating with media and H2O2 for 20 hours) and EGB group (adding media, 100μmol/L EGB and 100 μmol/L H2O2). ③ MTT assay: PC12 cells (1 × 10^8L^-1) were plated in 96-well plates and divided into three groups with 8 holes for each group. Under sterile condition, cells were added with 5 g/L MTT (100μL) and cultured for 4 hours. And then, 200 μ L DMSO fluid was added and shaken for 30 minutes until blue crystal products formed were dissolved soundly ④ Experimental evaluation: Absorbance (A) at 630 nm was measured a
关 键 词:PC12 cell H2O2: extracts of ginkgo biloba leaves
分 类 号:R329.28[医药卫生—人体解剖和组织胚胎学]
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