CAJ | 학술논문

目的应用高密度寡核苷酸(Oligo)基因芯片技术研究槲皮素对缺血缺氧损伤的星形胶质细胞基因表达的影响.方法体外原代培养星形胶质细胞分为缺血缺氧组和缺血缺氧+槲皮素处理组.2组细胞厌氧培养4h后缺血缺氧+槲皮素处理组加入含50 μmol/L槲皮素的培养液,缺血缺氧组加入等量培养液,培养24 h后应用基因表达谱芯片筛选2组细胞表达差异的基因并用实时荧光定量PCR检测进行验证.结果基因表达谱芯片分析显示缺血缺氧+槲皮素处理组与缺血缺氧组比较表达差异的基因共180个,其中上调基因49个,下调基因131个；实时荧光定量PCR结果显示缺血缺氧+槲皮素处理组细胞与缺血缺氧组比较148个基因的表达发生变化,差异均有统计学意义(P＜0.05),其中上调基因34个,下调基因114个.实时荧光定量PCR与基因表达谱芯片结果的符合率为82.2％(148/180).结论基因表达谱芯片分析有助于从分子水平全面了解槲皮素对缺血缺氧的星形胶质细胞的作用机制,也为进一步研究槲皮素和星形胶质细胞在缺血缺氧脑损伤中的作用奠定了基础.
목적 응용고밀도과핵감산(Oligo)기인심편기술연구곡피소대결혈결양손상적성형효질세포기인표체적영향.방법 체외원대배양성형효질세포분위결혈결양조화결혈결양+곡피소처리조.2조세포염양배양4h후결혈결양+곡피소처리조가입함50 μmol/L곡피소적배양액,결혈결양조가입등량배양액,배양24 h후응용기인표체보심편사선2조세포표체차이적기인병용실시형광정량PCR검측진행험증.결과 기인표체보심편분석현시결혈결양+곡피소처리조여결혈결양조비교표체차이적기인공180개,기중상조기인49개,하조기인131개；실시형광정량PCR결과 현시결혈결양+곡피소처리조세포여결혈결양조비교148개기인적표체발생변화,차이균유통계학의의(P＜0.05),기중상조기인34개,하조기인114개.실시형광정량PCR여기인표체보심편결과 적부합솔위82.2％(148/180).결론 기인표체보심편분석유조우종분자수평전면료해곡피소대결혈결양적성형효질세포적작용궤제,야위진일보연구곡피소화성형효질세포재결혈결양뇌손상중적작용전정료기출.
Objective To study the effect of quercetin on gene expression in glucose-oxygen deprived astrocytes using large-scale oligo microarray technology.Methods Astrocytes were primarily cultured in vitro and divided into ischemia and hypoxia group and ischemia and hypoxia plus quercetin treatment(50 μmol/L)group; ischemia and hypoxia cells from these 2 groups were induced by anaerobic culture for 4 h,and then,the nutrient solutions were added into each group,respectively.Twenty-four h after that,cDNA microarray was employed to select the differentially expressed genes in the 2 groups,and these genes were confirmed by quantitative RT-PCR.Results The cDNA microarray indicated that the expressions of 180 genes had significant changes at the mRNA level between ischernia and hypoxia group and ischemia and hypoxia plus quercetin treatment group,of which 49 genes were up-regulated and 131 were down-regulated.One hundred and forty-eight differentially expressed genes were confirmed by RT-PCR,including 34 up-regulated genes and 114 down-regulated genes,which showed that 82.2％(148/180)genes that matched with the results of cDNA microarray.Conclusion Gene expression profiling by large-scale oligo microarray provides good understanding of the molecular mechanism of quercetin in glucose-oxygen deprived astrocytes,and laids the foundation for investigating the influence of quercetin and astrocytes in hypoxic-ischemic brain damage.