Effect of Norathyriol on the Epithelial-mesenchymal Transition of HK-2 Cells Induced by TGF-β1
-
摘要:
目的 研究芒果苷元对TGF-β1诱导的HK-2细胞EMT的影响。 方法 MTT法检测各浓度芒果苷元对HK-2细胞活力的影响；用TGF-β1诱导HK-2细胞成EMT模型，并给以芒果苷元干预，在TGF-β1诱导24 h时显微镜下拍照记录细胞形态变化，划痕实验法检测细胞迁移能力，Transwell法检测细胞侵袭能力，Western blot法检测纤连蛋白（fibronectin，FN）和Ⅰ型胶原（collagen type Ⅰ，Col Ⅰ）蛋白表达水平。 结果 （1）芒果苷元（10-9～10-5 mol/L）浓度下HK-2细胞活力均无明显变化（P > 0.05）；（2）与正常组相比，模型组细胞形态变长变梭，迁移和侵袭能力显著增强（ P < 0.05，0.01），FN和ColⅠ蛋白表达水平显著上调（ P < 0.05，0.01）；与模型组相比，芒果苷元组细胞能维持原来的鹅卵石形，迁移和侵袭能力显著下降（ P < 0.05，0.01），FN和ColⅠ蛋白表达水平显著下调（ P < 0.05）。 结论 芒果苷元能抑制TGF-β1诱导的HK-2细胞迁移和侵袭，阻止细胞形态的改变，下调细胞外基质成分FN和ColⅠ的蛋白表达，从而抑制EMT的发展。 Abstract:Objective To study the effects of norathyriol on epithelial-mesenchymal transition of HK-2 cells induced by TGF-β1. Methods MTT assay was used to detect the effect of norathyriol on HK-2 cell viability. The HK-2 cells were divided into normal group, model group, norathyriol （10-8, 10-7, 10-6 mol·L-1） groups and positive control （benzbromarone, 10-7 mol/L） group. The norathyriol and benzbromarone were incubated for 4 h before HK-2 cells were treated by TGF-β1 （10 ng/mL） to induce EMT. Then, the HK-2 cells were cultured for another 24 h or 48 h. Scratch assay was used to detect the migration motility of HK-2 cells. Transwell invasion assay was used to detect invasion ability of HK-2 cells. The proteins expression of fibronectin （FN） and collagen type Ⅰ （ColⅠ） were detected by western blot. Results （1） The norathyriol （10-9-10-5 mol/L）has no effects on the HK-2 cell viability, compared with the normal group （P > 0.05）. （2） Compared with the normal group, the cells morphology of the model group changed from the original cobblestone shape to a long fusiform shape, the migration rate and the invasive ability of the model group was significantly increased （ P < 0.01）. The protein expression of Col and FN in the model group was significantly increased. Compared with the model group, the cells morphology of norathyriol （10 -8, 10-7, 10-6 mol/L） and benzbromarone （10-7 mol/L） groups maintain thecobblestone shape partly, the migration rate （10-8, 10-7, 10-6 mol/L） and the invasive ability （10-6 mol/L） of norathyriol were significantly decreased （P < 0.01）; The protein expression of Col and FN were significantly down-regulated in the norathyriol （10 -6 mol/L） group and the benzbromarone group （P < 0.05, 0.01）. Conclusion Norathyriol can inhibit the migration and invasion of HK-2 cells induced by TGF-β1 and down-regulate the protein expression of fibronectin （FN） and collagen type Ⅰ （ColⅠ）. -
Key words:
- Norathyriol /
- TGF-β1 /
- HK-2 cell /
- Migration /
- Invasion /
- Epithelial-mesenchymal transition （EMT）
-
图 1 芒果苷元对HK-2细胞活力的影响（ $\bar x \pm s$，n = 3）
注：NOR：正常组；NL：芒果苷元；Ben：苯溴马隆。
Figure 1. Effect of norathyriol on HK-2 cell viability
图 2 芒果苷元对TGF-β1 诱导24 h后的HK-2细胞形态变化的影响（40×）
注：NOR：正常组；MOD：模型组；NL：芒果苷元；Ben：苯溴马隆。
Figure 2. Effect of norathyriol on the morphological changes of HK-2 cells induced by TGF - β 1 for 24 h （40×）
图 3 芒果苷元对TGF-β1诱导后HK-2细胞迁移率的影响（40×）
Figure 3. Effect of norathyriol on the migration of HK-2 cells induced by TGF - β 1 （40×）
图 4 芒果苷元对TGF-β1诱导后的HK-2细胞的迁移率的影响（ $\bar x \pm s$，n = 3）
注：NOR：正常组；MOD：模型组；NL：芒果苷元；Ben：苯溴马隆。与模型组相比，*P < 0.05；**P < 0.01；与正常组相比，##P < 0.01。
Figure 4. Effect of norathyriol on the migration of HK-2 cells induced by TGF – β1 （ $\bar x \pm s$，n = 3）
图 5 芒果苷元对TGF-β1诱导的HK-2细胞侵袭能力的影响（100×）
Figure 5. Effect of norathyriol on invasion of HK-2 cells induced by TGF - β 1（100 ×）
图 6 芒果苷元对TGF-β1诱导的HK-2细胞侵袭能力的影响（ $\bar x \pm s$，n = 3）
注：NOR：正常组；MOD：模型组；NL：芒果苷元；Ben：苯溴马隆。与模型组相比，*P < 0.05；与正常组相比，##P < 0.01，one-way ANOVA。
Figure 6. Effect of norathyriol on invasion of HK-2 cells induced by TGF - β 1 （ $\bar x \pm s$，n = 3）
图 7 芒果苷元对TGF-β1诱导的HK-2细胞ColⅠ蛋白表达水平的影响（ $\bar x \pm s$，n = 3）
注：NOR：正常组；MOD：模型组；NL：芒果苷元；Ben：苯溴马隆。与正常组相比，#P < 0.05；与模型组比，*P < 0.05，**P < 0.01，one-way ANOVA。
Figure 7. Effect of norathyriol on expression of colⅠprotein in HK-2 cells induced by TGF-β1 （ $\bar x \pm s$，n = 3）
-
[1] Liu B C,Tang T T,Lv L L,et al. Renal tubule injury:A driving force toward chronic kidney disease[J]. Kidney international,2018,93(3):568-579. doi: 10.1016/j.kint.2017.09.033 [2] Djudiai S,Boor P. Cellular and molecular mechanisms of kidney fibrosis[J]. Molecular Aspects of Medicine,2019,65:16-36. doi: 10.1016/j.mam.2018.06.002 [3] 石咏琪,王墨. 肾间质纤维化发生机制研究进展[J]. 现代医药卫生,2019,35(19):3002-3006. doi: 10.3969/j.issn.1009-5519.2019.19.020 [4] Cruz-Solbes A S,Youker K. Epithelial to mesenchymal transition (EMT) and endothelial to mesenchymal transition (EndMT):role and implications in kidney fibrosis[J]. Results and Problems in Cell Differentiation,2017,60:345-372. [5] Iwano M,Plieth D,Danoff T M,et al. Evidence that fibroblasts derive from epithelium during tissue fibrosis[J]. The Journal of Clinical Investigation,2002,110(3):341-350. doi: 10.1172/JCI0215518 [6] 遆安航,陈瑾,翟康欣,等. 连翘归尾煎对人乳腺癌MCF-7细胞增殖、迁移、凋亡和周期的影响[J]. 山西医科大学学报,2021,52(5):565-571. [7] Saral L,Michael Z,Raghu K. Partial epithelial-to-mesenchymal transition and other new mechanisms of kidney fibrosis[J]. Trends in Endocrinology and Metabolism,2016,27(10):681-695. doi: 10.1016/j.tem.2016.06.004 [8] Meng X M,Nikolic-paterson D J,Lan H Y. TGF-β:the master regulator of fibrosis[J]. Nature Reviews. Nephrology,2016,12(6):325-338. doi: 10.1038/nrneph.2016.48 [9] Xu C G,Zhu X L,Wang W,et al. Ursolic acid inhibits epithelial-mesenchymal transition in vitro and in vivo[J]. Pharmaceutical Biology,2019,57(1):169-175. doi: 10.1080/13880209.2019.1577464 [10] Fan Y,Wang X,Li Y,et al. PAK4 enhances TGF-β1-induced epithelial- mesenchymal transition through activating β-catenin signaling pathway in renal tubular epithelial cells[J]. International Journal of Clinical and Experimental pathology,2018,11(6):3026-3035. [11] Song Y,Lv S S,Wang F,et al. Overexpression of BMP-7 reverses TGF-β1-induced epithelial-mesenchymal transition by attenuating the Wnt3/β-catenin and /Smad2/3 signaling pathways in HK-2 cells[J]. Molecular Medicine Reports,2019,21(2):833-841. [12] 周栋,傅海燕. 细胞外基质的产生、重塑与肾脏纤维化[J]. 江苏医药,2015,41(8):936-938. [13] 李淑菊,詹晓丹,田锋,等. 参地补肾胶囊含药血清对高糖诱导转分HK-2细胞ZO-1、α-SMA表达的影响[J]. 中国中医药科技,2018,25(6):797-800. [14] Louis K,Hertig A. How tubular epithelial cells dictate the rate of renal fibrogenesis?[J]. World Journal of Nephrology,2015,4(3):367-373. doi: 10.5527/wjn.v4.i3.367 -
下载:
点击查看大图
计量
- 文章访问数: 4063
- HTML全文浏览量: 2226
- PDF下载量: 128
- 被引次数: 0
