Effect of Sedum sarmentosum Bunge Extract on TGF-β1-induced renal epithelial-to-mesenchymal transition and matrix accumulation
1.Department of Laboratory Medicine, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325015; 2.Wenzhou Key Laboratory of Surgery, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325015
Abstract：Objective: To investigate the protective effects of Sedum sarmentosum Bunge Extract (SSBE) on TGF-β1-induced epithelial-to-mesenchymal transition (EMT) and matrix accumulation in renal tubular epithelial cells (NRK-52E). Methods: NRK-52E cells were divided randomly into: control group treated with only solvent, TGF-β1 group treated with TGF-β1 at the concentrations with 5 µg/L and SSBE group treated with TGF-β1 plus SSBE at the concentrations with 10 and 100 mg/L. The morphology of the NRK-52E cells was observed under inverted/phase contrast microscope. Immunofluorescent analysis was performed to detect the expression of epithelial marker α-smooth muscle actin (α-SMA), mesenchymal marker E-cadherin, and matrix component type III collagen. Gene expression of α-SMA, bone morphogenic protein-7 (BMP-7), tight junction protein-1 (Tjp1), Col1a1, Col3a1, MMP-2, and TIMP-2 were also quantified by real-time RT-PCR. Results: In TGF-β-treated NRK-52E cells, fibrosis-like phonotype was obviously increased. TGF-β1 increased the expression of α-SMA, and decreased the expression of E-cadherin and Tjp1. Also, TGF-β1 enhanced the expression of type I and III collagens. Treatment with SSBE at the concentrations with 10 and 100 mg/L inhibited TGF-β1-induced fibrosis-like phenotype of NRK-52E cells, accompanied with down-regulated expression of α-SMA, Col1a1 and Col1a1 and up-regulated expression of E-cadherin and Tip1. In addition, SSBE increased the expression of BMP-7 and the ratio between MMP-2 and TIMP-2. Conclusion: SSBE treatment reduce TGF-β1-induced fibrosis-like reaction in renal tubular epithelial cells through inhibiting EMT and matrix accumulation.
[1] Liu Y. Cellular and molecular mechanisms of renal fibrosis [J]. Nat Rev Nephrol, 2011, 7(12): 684-696.
[2] Liu Y. New insights into epithelial-mesenchymal transition in kidney fibrosis[J]. J Am Soc Nephrol, 2010, 21(2): 212-222.
[3] Bai Y, Lu H, Hu L, et al. Effect of Sedum sarmentosum BUNGE extract on aristolochic acid-induced renal tubular epithelial cell injury[J]. J Pharmacol Sci, 2014, 124(4): 445-
456.
[4] Xu Y, Wan J, Jiang D, et al. BMP-7 counteracts TGF-beta1-induced epithelial-to-mesenchymal transition in human renal proximal tubular epithelial cells[J]. J Nephrol, 2009, 22(3):403-410.
[5] Anglani F, Ceol M, Mezzabotta F, et al. The renal stem cell system in kidney repair and regeneration[J]. Front Biosci,2008, 13: 6395-6405.
[6] 胡丽萍, 陆红, 白永恒, 等. 小G蛋白Rac1在马兜铃酸诱导肾小管上皮细胞损伤中的作用及意义[J]. 温州医学院学报, 2013, 43(12): 775-778.
[7] Lian YG, Zhou QG, Zhang YJ, et al. VEGF ameliorates tubulointerstitial fibrosis in unilateral ureteral obstruction mice via inhibition of epithelial-mesenchymal transition[J]. Acta Pharmacol Sin, 2011, 32(12): 1513-1521.
[8] Bai Y, Lu H, Zhang G, et al. Sedum sarmentosum Bunge extract exerts renal anti-fibrotic effects in vivo and in vitro[J]. Life Sci, 2014, 105(1-2): 22-30.
[9] Yoon JS, Chae MK, Jang SY, et al. Antifibrotic effects of quercetin in primary orbital fibroblasts and orbital fat tissue cultures of Graves’ orbitopathy[J]. Invest Ophthalmol Vis Sci, 2012, 53(9): 5921-5929.
