Guo Wei, Xu Hong, Chen Jing, Yang Yong, Jin Jia Wei, Fu Rui, Liu Hai Mei, Zha Xi Liang, Zhang Zhi Gang, Huang Wen Yan
Department of Nephrology, Children's Hospital of Fudan University, 183 Feng Lin Road, Shanghai, 200032, P.R. China.
Mol Cell Biochem. 2007 Jan;295(1-2):167-77. doi: 10.1007/s11010-006-9286-4. Epub 2006 Oct 17.
Prohibitin (PHB), a potential tumor suppressor, has been shown to inhibit cell proliferation by repressing E2F-mediated transcription. But little is known about the role of PHB involved in tubulointerstitial fibrosis (TIF). Here, for the first time, we found PHB protein was positively expressed at normal renal tissues, strongly down-regulated in renal biopsy specimens, and negatively correlated with the expression of alpha-smooth-muscle actin (alpha-SMA) and with the degrees of tubulointerstitial lesions. Transforming growth factor-beta1 (TGF-beta1) is the most important profibrotic cytokine in the process of TIF and capable of inducing cell phenotypic change of interstitial fibroblasts characterized by the de novo expression of alpha-SMA. Confocal microscopy showed majority of PHB is located at cytoplasm as well as at nucleus in rat kidney fibroblasts cell (NRK-49F). As we found that PHB protein and mRNA expression were down-regulated in NRK-49F cells following TGF-beta1 stimulation. We used transient transfection to over-express PHB protein and found that cells with increased PHB levels had a significant reduction in the percentage entering cell cycle and abolished de novo expression of alpha-SMA following TGF-beta1 stimulation. Therefore, over-expression of PHB suppresses renal interstitial fibroblasts proliferation and cell phenotypic change induced by TGF-beta1, which indicates PHB as a potential therapeutic target to halt the progression of TIF.
抑制素(PHB)是一种潜在的肿瘤抑制因子,已被证明可通过抑制E2F介导的转录来抑制细胞增殖。但关于PHB在肾小管间质纤维化(TIF)中的作用知之甚少。在此,我们首次发现PHB蛋白在正常肾组织中呈阳性表达,在肾活检标本中强烈下调,且与α-平滑肌肌动蛋白(α-SMA)的表达及肾小管间质病变程度呈负相关。转化生长因子-β1(TGF-β1)是TIF过程中最重要的促纤维化细胞因子,能够诱导以α-SMA从头表达为特征的间质成纤维细胞表型改变。共聚焦显微镜显示,在大鼠肾成纤维细胞(NRK-49F)中,大多数PHB位于细胞质以及细胞核中。我们发现,在TGF-β1刺激后,NRK-49F细胞中PHB蛋白和mRNA表达下调。我们通过瞬时转染使PHB蛋白过表达,发现PHB水平升高的细胞进入细胞周期的百分比显著降低,并且在TGF-β1刺激后α-SMA的从头表达被消除。因此,PHB的过表达抑制了肾间质成纤维细胞的增殖以及TGF-β1诱导的细胞表型改变,这表明PHB是阻止TIF进展的潜在治疗靶点。