microRNA-132在人臍靜脈內皮細胞中調控作用的研究

張櫪心，陶利娟

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·實驗論著·

microRNA-132在人臍靜脈內皮細胞中調控作用的研究

張櫪心，陶利娟

Department of Ophthalmology,Hunan Children’s Hospital,Changsha 410007,Hunan Province,China

Abstract

?AIM: To evaluate the regulatory effect of microRNA-132 (miR-132) in human umbilical vein endothelial cell (HUVEC).

?METHODS: In vitro cultured human umbilical vein endothelia cells in hypoxic environment for 6h,then maintained under normal oxygen condition for 3h,6h,12h,24h.miR-132 and peroxisome-proliferator-activated receptor-γ coactivator-1α (PGC-1α) expression was detected by quantitative Real-time polymerase chain reaction and Western blot analysis.Human umbilical vein endothelial cells transfected miR-132 mimic and miR-132 inhibitor(anti-miR-132) were measured by quantitative Real-time polymerase chain reaction and Western blot.

?RESULTS: miR-132 and PGC-1α expression was significantly (P<0.01) upregulated in the hypoxic environment of cells at 3h compared with the normal oxygen condition.After cells transfection,the hypoxic environment the miR-132 and PGC-1α expression were markedly increased compared with the normal oxygen condition.The cells transfected miR-132-mimic,the expression of the miR-132 and PGC-1α were higher than that of transfected anti-miR-132 and contrast group (P<0.01).

?CONCLUSION: miR-132 level is highly expressed in the HUVEC under hypoxia and may be an effect of regulation for PGC-1α.

目的：探討miR-132在人臍靜脈內皮細胞(human umbilical vein endothelia cells，HUVEC)中的調控作用。

方法：體外低氧培養人臍靜脈內皮細胞6h后繼續常氧培養3、6、12、24h，利用Real-time PCR檢測miR-132和PGC-1α mRNA表達變化，Western-blot檢測PGC-1α 蛋白表達變化，及觀察各組與正常培養的細胞之間的表達差異。通過對人臍靜脈內皮細胞轉染miR-132模擬物與拮抗劑后，再分別置于常氧和低氧環境中培養，利用Real-time PCR檢測不同氧條件下miR-132和PGC-1α mRNA的表達變化，Western-blot檢測PGC-1α蛋白表達變化。

結果：miR-132和PGC-1α在細胞低氧培養后繼續常氧培養3h時蛋白與mRNA表達量最高，與正常培養的細胞表達差異最為明顯(P<0.01)。細胞轉染后可觀察到miR-132和PGC-1α在低氧組表達量要高于常氧組，而兩組中轉染miR-132模擬物后的表達量要高于轉染拮抗劑組(P<0.01)。

結論：miR-132在低氧時的人臍靜脈內皮細胞中表達明顯上調，對PGC-1α存在調控作用。

microRNA-132；過氧化物酶增殖激活受體-γ共激活子-1α；人臍靜脈內皮細胞；缺氧

引用：張櫪心，陶利娟.microRNA-132在人臍靜脈內皮細胞中調控作用的研究.國際眼科雜志2016;16(10):1820-1823

0　引言

微小RNA(microRNA，miRNA)是一類生物內源的非編碼小RNA，由20～24個核苷酸組成，通過與mRNA轉錄物的3’非翻譯區結合而調控基因轉錄后表達[1]。miR-132是發現較早的miRNA之一，2002年馬克斯普朗克學會的Lagos-Quintana等首次在小鼠神經組織中發現miR-132[2]，成熟的miR-132長度為22bp，由長度為66bp的前體序列進行加工而成[3]，miR-132具有進化保守性，在人、大鼠、小鼠、猿等物種中具有相同的序列與結構。作為內皮細胞特異性miRNA之一，miR-132在內皮靶向p120RasGAP誘導新血管形成被稱為血管生成開關。調查結果顯示，在人類血管生成的胚胎干細胞模型中，miR-132高度上調，這在正常內皮中是檢測不到的[4]。前期研究……