Li Yongjian, Zu Xuyu, Zhang Guiying, Xiao Rong, Wen Haiquan
Department of Dermatology, First Affiliated Hospital of Nanhua University, Hengyang Hunan, China.
Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2011 Oct;36(10):1012-6. doi: 10.3969/j.issn.1672-7347.2011.10.014.
To observe the effect of hypoxia inducible factor -1α (HIF-1α) small interfering RNA (siRNA) on the expression of HIF-1α and inducible nitric oxide synthase (iNOS) in HaCaT cells under hypoxia.
HaCaT cells were divided into 4 groups: the normal control group (without any treatment), the hypoxia group (under hypoxia for 24 h), the liposome control group (HaCaT cells transfected with liposome before hypoxia treatment), the RNA interference group (HaCaT cells transfected with siRNA sequences then under hypoxia for 24 h). Real-time PCR and Western blot were utilized to determine HIF-1α and iNOS mRNA and protein expression in HaCaT cells.
There was no significant difference of the mRNA expression of HIF-1α between the hypoxia group and the normoxia group (P>0.05), but the protein expressions of HIF-1α was increased in the hypoxic group than that in the normoxia group (P<0.05). Both the mRNA and protein expression of iNOS were increased in hypoxic conditions than that in the normoxia (P<0.05). Decreases were more significant in the mRNA and protein expression of HIF-1α and iNOS in the RNA interference group than that in the liposome control group in HaCaT cells (P<0.05).
Hypoxia increased HIF-1α and iNOS expression in HaCaT cells and inhibition of HIF-1α expression decreased iNOS expression.
观察缺氧诱导因子-1α（HIF-1α）小干扰RNA（siRNA）对缺氧条件下HaCaT细胞中HIF-1α及诱导型一氧化氮合酶（iNOS）表达的影响。
将HaCaT细胞分为4组：正常对照组（未作任何处理）、缺氧组（缺氧24小时）、脂质体对照组（缺氧处理前用脂质体转染HaCaT细胞）、RNA干扰组（转染siRNA序列后缺氧处理24小时）。采用实时荧光定量PCR和蛋白质免疫印迹法检测HaCaT细胞中HIF-1α和iNOS的mRNA及蛋白表达。
缺氧组与常氧组HIF-1α的mRNA表达差异无统计学意义（P>0.05），但缺氧组HIF-1α蛋白表达高于常氧组（P<0.05）。缺氧条件下iNOS的mRNA和蛋白表达均高于常氧组（P<0.05）。HaCaT细胞中，RNA干扰组HIF-1α和iNOS的mRNA及蛋白表达较脂质体对照组降低更显著（P<0.05）。
缺氧可增加HaCaT细胞中HIF-1α和iNOS的表达，抑制HIF-1α表达可降低iNOS表达。
