| 摘要: |
| [目的] 探究栀子苷（GE）对高糖诱导的肾小管上皮细胞转分化中转化生长因子-β和Smad相关蛋白（TGF-β/Smad）通路的影响。[方法] 实验分为对照组、高糖组、GE 1、10、50、100 μmol/L组。对照组HK2细胞在5.5 mmol/L低糖DMEM培养液中培养，除对照组外，其余各组均于25 mmol/L高糖DMEM培养液中培养，GE 1、10、50、100 μmol/L组同时添加GE，使GE终浓度分别为1、10、50、100 μmol/L，干预48 h。显微镜下观察HK2细胞形态；实时荧光定量聚合酶链反应（qRT-PCR）检测细胞中纤连蛋白（FN）、E-钙黏素（E-cad）、Ⅳ型胶原（COLⅣ）mRNA水平；酶联免疫吸附法（ELISA）检测上清液中白细胞介素（IL）-1β、IL-6、肿瘤坏死因子-α （TNF-α）水平；蛋白免疫印迹法（Western blot）检测细胞中转化生长因子-β1（TGF-β1）、Smad3、磷酸化Smad3 （p-Smad3）、α-平滑肌肌动蛋白（α-SMA）蛋白水平。[结果] 对照组细胞呈多边形或卵圆形，分布均匀；高糖组细胞部分呈现长梭形、胞核呈梭形变；随着GE剂量的升高，细胞形态逐渐恢复正常。与对照组相比，高糖组细胞中FN、COLⅣ mRNA水平，TGF-β1、p-Smad3/Smad3、α-SMA蛋白水平，上清液中IL-1β、IL-6、TNF-α水平升高，细胞中E-cad mRNA水平降低（P＜0.05）；与高糖组相比，GE 1 μmol/L组细胞中FN mRNA水平，TGF-β1、p-Smad3/Smad3、α-SMA蛋白水平，上清液中IL-1β、IL-6、TNF-α水平降低（P＜0.05），GE 10、50、100 μmol/L组细胞中FN、COLⅣ mRNA水平，TGF-β1、p-Smad3/Smad3、α-SMA蛋白水平，上清液中IL-1β、IL-6、TNF-α水平降低，细胞中E-cad mRNA水平升高（P＜0.05）。[结论] GE可以抑制高糖诱导的肾小管上皮细胞转分化中TGF-β/Smad通路。 |
| 关键词: 栀子苷 肾小管上皮细胞 转分化 转化生长因子-β和Smad相关蛋白通路 |
| DOI：10.11656/j.issn.1672-1519.2022.08.21 |
| 分类号:R285.5 |
| 基金项目:天津市教委科研计划项目（2017KJ154）。 |
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| Effect of geniposide on TGF-β/Smad pathway in high glucose-induced trans-differentiation of renal tubular epithelial cells |
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LIANG Dong1,2, YANG Hongtao1,2
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1.First Teaching Hospital of Tianjin University of Traditional Chinese Medicine, Tianjin 300381, China;2.National Clinical Research Center for Chinese Medicine Acupuncture and Moxibustion, Tianjin 300381, China
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| Abstract: |
| [Objective] To investigate the effects of geniposide (GE) on transforming growth factor-β and Smad related protein (TGF-β/ Smad) pathway in high glucose-induced trans-differentiation of renal tubular epithelial cells. [Methods] The experiment was divided into control group,high glucose group and GE 1,10,50,100 μmol/L groups. HK2 cells in the control group were cultured in 5.5 mmol/L low glucose DMEM medium. Except the control group,the other groups were cultured in 25 mmol/l high glucose DMEM medium,GE 1 ,10,50,100 μmol/L groups were added with GE at the same time,and the final concentration of GE was 1,10,50, 100 μmol/L respectively for 48 h. The morphology of hK2 cells was observed under microscope,the mRNA levels of fibronectin (FN),E-cadherin (E-cad) and collagen type Ⅳ (COLⅣ) were detected by real-time fluorescence quantitative PCR (qRT-PCR),the levels of interleukin (IL)-1β,IL-6 and tumor necrosis factor-α (TNF-α) in the supernatant were detected by enzyme-linked immunosorbent assay (ELISA), the protein levels of TGF-β1,Smad3,and α-smooth muscle actin (α-SMA) were detected by Western blot. [Results] The cells in the control group were polygonal or oval in shape and evenly distributed；in the high glucose group,some of the cells were spindle shaped and the nucleus was spindle shaped,with the increase of GE dose,the cell morphology gradually returned to normal. Compared with those in the control group,the mRNA levels of FN and COLⅣ,the protein levels of TGF-β1,p-Smad3/Smad3,α-SMA protein,and the levels of IL-1β,IL-6 and TNF-α in the supernatant in high glucose group were higher,and the mRNA level of E-cad was lower (P<0.05). Compared with those in the high glucose group,the mRNA level of FN,the protein levels of TGF-β1,p-Smad3/Smad3,α-SMA protein,and the levels of IL-1β,IL-6 and TNF-α in the supernatant in GE 1 μmol/L group were lower (P<0.05),in GE (10,50,100) μmol/L group, the mRNA levels of FN and COLⅣ,the protein levels of TGF-β1,p-Smad3/Smad3,α-SMA protein,and the levels of IL-1β,IL-6 and TNF-α in the supernatant were lower,and the mRNA level of E-cad in the cells was higher (P<0.05). [Conclusion] GE can inhibit TGF- β/Smad pathway in high glucose-induced trans-differentiation of renal tubular epithelial cells. |
| Key words: geniposide renal tubular epithelial cell trans-differentiation transforming growth factor-β and Smad related protein pathway |
