| 摘要: |
| [目的]建立人肝星状细胞（LX-2）与人肾小管上皮细胞（HK-2）共培养体系的理论模型，研究LX-2对HK-2转分化的影响。[方法]以Transwell小室建立体外LX-2细胞和HK-2细胞间接共培养体系。应用细胞免疫组织化学法检测HK-2细胞平滑肌肌动蛋白（α-SMA）表达；用酶联免疫吸附法（ELISA法）检测各组细胞培养液中转化生长因子-β1（TGF-β1）的含量；逆转录-聚合酶链反应法（RT-PCR法）检测HK-2细胞TβRⅠmRNA、TβRⅡmR-NA、Smad2 mRNA、Smad3 mRNA、Smad7 mRNA的表达。[结果]除了空白组，形态学上其他两组均可看到HK-2细胞胞浆染棕黄色，不同数量细胞由立方铺路石样转变为梭形长条状，细胞肥大；HK-2细胞高表达α-SMA（P<0.01）；HK-2细胞上清液中TGF-β1含量明显高于空白组（P<0.01）；HK-2细胞TβRⅠ、Smad2、Smad3基因表达明显上调，而TβRⅡ、Smad7基因表达明显下调。[结论]Transwell共培养法可诱导HK-2细胞转分化，其机制可能与TGF-β1/Smad信号通路有关。 |
| 关键词: 肾小管上皮细胞转分化 Transwell小室 LX-2细胞 HK-2细胞 细胞共培养 |
| DOI：10.11656/j.issn.1673-9043.2016.05.09 |
| 分类号: |
| 基金项目:天津市教委课题项目（20070307）。 |
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| LX-2 and HK-2 cell co-culture model and the influence of its cells |
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WANG Yao-guang1, ZENG You-jia2, ZHOU Hui-jie3
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1.First Teaching Hospital of Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China;2.Shenzhen City Hospital of traditional Chinese medicine, Shenzhen 518033, China;3.Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China
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| Abstract: |
| [Objective] To establish human hepatic stellate cells (LX-2) and renal tubular epithelial cells (HK-2) co-culture system theory model, research LX-2 cells influence on transdifferentiation of HK-2 cells.[Methods] By Transwell Chambers establish human hepatic stellate cells in vitro (LX-2 cells) and human renal tubular epithelial cells (HK)-2 cells indirectly co-culture system. By cellular immune histochemical method to detect α-SMA of HK-2 cells; With ELISA method to detect the TGF-beta 1 in cell cultures of the content; By RT-PCR method to detect of T beta RⅠmRNA, T beta R Ⅱ mRNA, Smad2 mRNA, Smad3 mRNA and Smad7 mRNA expression of HK-2 cell.[Results] Except the blank group, the two other groups on morphology can see HK-2 dye cell cytoplasm tan, different number of cells from cubic paving stone sample into spindle long strips, cell hypertrophy; HK-2 high expression of α-SMA cells (P<0.01); TGF-beta 1 content in the HK-2 cells supernatant fluid was obviously higher than blank group (P<0.01). TβRⅠ, Smad2 and Smad3 gene expression increase obviously, while TβRⅡ, Smad7 gene expression significantly lowered in HK-2 cells.[Conclusion] Transwell culture method can be induced to HK-2 cell trans-differentiation, its mechanism may be related to the TGF beta 1/Smad signaling pathways. |
| Key words: renal tubular epithelial cell trans-differentiation Transwell LX-2 cell HK-2 cell cell co-culture |
