中华老年心脑血管病杂志
中華老年心腦血管病雜誌
중화노년심뇌혈관병잡지
CHINESE JOURNAL OF GERIATRIC CARDIOVASCULAR AND CEREBROVASCULAR DISEASES
2014年
6期
633-636
,共4页
艾丽菲热·买买提%陈红%任景怡
艾麗菲熱·買買提%陳紅%任景怡
애려비열·매매제%진홍%임경이
动脉粥样硬化%微RNAs%转染%内皮细胞%细胞凋亡%多基因族%转录激活因子3
動脈粥樣硬化%微RNAs%轉染%內皮細胞%細胞凋亡%多基因族%轉錄激活因子3
동맥죽양경화%미RNAs%전염%내피세포%세포조망%다기인족%전록격활인자3
atherosclerosis%microRNAs%transfection%endothelial cells%apoptosis%multigene fami-ly%activating transcription factor 3
目的：研究微小RNA（microRNA ，miR）-106b是否参与动脉粥样硬化相关的血管新生，明确miR-106b在内皮细胞中参与血管新生的作用机制。方法内皮细胞培养并转染miR-106b ，分为miR-106b组、空白对照组和阴性对照组。提取RNA、反转录及实时定量PCR检测miR-106b相对表达量以明确转染效率。观察各组内皮细胞在凝固的基质胶中管腔形成情况。TUNEL法检测转染后细胞凋亡状况并进行靶基因的预测，采用Western blot法检测筛选的蛋白相对表达量变化。结果与空白对照组和阴性对照比较，miR-106b组在基质胶中管腔形成明显减少；与阴性对照组比较，miR-106b组管腔比值、信号转导及转录激活因子3 mRNA相对表达量及蛋白表达明显降低（P＜0．05，P＜0．01）。阴性对照组与miR-106b组细胞凋亡率比较，差异无统计学意义（1．19％ vs 3．39％，P＞0．05）。结论 miR-106b在内皮细胞抑制血管新生可能是通过下调信号转导及转录激活因子3，与血管内皮生长因子无直接关系。
目的：研究微小RNA（microRNA ，miR）-106b是否參與動脈粥樣硬化相關的血管新生，明確miR-106b在內皮細胞中參與血管新生的作用機製。方法內皮細胞培養併轉染miR-106b ，分為miR-106b組、空白對照組和陰性對照組。提取RNA、反轉錄及實時定量PCR檢測miR-106b相對錶達量以明確轉染效率。觀察各組內皮細胞在凝固的基質膠中管腔形成情況。TUNEL法檢測轉染後細胞凋亡狀況併進行靶基因的預測，採用Western blot法檢測篩選的蛋白相對錶達量變化。結果與空白對照組和陰性對照比較，miR-106b組在基質膠中管腔形成明顯減少；與陰性對照組比較，miR-106b組管腔比值、信號轉導及轉錄激活因子3 mRNA相對錶達量及蛋白錶達明顯降低（P＜0．05，P＜0．01）。陰性對照組與miR-106b組細胞凋亡率比較，差異無統計學意義（1．19％ vs 3．39％，P＞0．05）。結論 miR-106b在內皮細胞抑製血管新生可能是通過下調信號轉導及轉錄激活因子3，與血管內皮生長因子無直接關繫。
목적：연구미소RNA（microRNA ，miR）-106b시부삼여동맥죽양경화상관적혈관신생，명학miR-106b재내피세포중삼여혈관신생적작용궤제。방법내피세포배양병전염miR-106b ，분위miR-106b조、공백대조조화음성대조조。제취RNA、반전록급실시정량PCR검측miR-106b상대표체량이명학전염효솔。관찰각조내피세포재응고적기질효중관강형성정황。TUNEL법검측전염후세포조망상황병진행파기인적예측，채용Western blot법검측사선적단백상대표체량변화。결과여공백대조조화음성대조비교，miR-106b조재기질효중관강형성명현감소；여음성대조조비교，miR-106b조관강비치、신호전도급전록격활인자3 mRNA상대표체량급단백표체명현강저（P＜0．05，P＜0．01）。음성대조조여miR-106b조세포조망솔비교，차이무통계학의의（1．19％ vs 3．39％，P＞0．05）。결론 miR-106b재내피세포억제혈관신생가능시통과하조신호전도급전록격활인자3，여혈관내피생장인자무직접관계。
Objective To study whether miR-106b is involved in endothelial cells-mediated angio-genesis .Methods miR-106b cultured and transfected with endothelial cells was divided into miR-106b group ,blank control group and positive control group .RNA was extracted from miR-106b .The transfection efficiency was confirmed by inverse transcription .Endothelial cell tubes formed in matrigel were observed .Apoptosis of transfected miR-106b was assayed by TUNEL assay .Target genes of miR-106b were detected .Expressions of miR-106b and candidate genes were detected by RT-PCR and Western blot ,respectively .Results The number of endothe-lial cell tubes formed in matrigel was significantly less in miR-106b group than in blank control group and positive control group .The ratio of formed tubes ,signal transduction and mRNA ex-pression level were significantly lower in miR-106b group than in positive control group ( P<0.05 ,P<0.01) .No significant difference was found in apoptosis of transfected miR-106b among the 3 groups (1.19% vs 3 .39% ,P>0 .05) .Conclusion miR-106b inhibits endothelial cells-medi-ated angiogenesis by down-regulating the signal transduction and STAT 3 ,which is not directly re-lated with VEGFA .
