摘要
目的探讨螺内酯对人清蛋白诱导的人近端肾小管上皮细胞(HK-2细胞)发生转分化的影响,以及该过程是否通过转化生长因子β1(TGF-β1)/Smads信号通路发挥作用。方法体外培养HK-2细胞,将其随机分为7组:A组(未加刺激物)、B组(加入10-7 mol/L螺内酯)、C组(加入5mg/mL清蛋白)、D组(加入5mg/mL清蛋白及10-8 mol/L螺内酯)、E组(加入5mg/mL清蛋白及10-7 mol/L螺内酯)、F组(加入5mg/mL清蛋白及10-6 mol/L螺内酯)、G组(先加10μmol/L TGF-β1拮抗剂,30min后加入5mg/mL清蛋白)。采用RT-PCR检测TGF-β1和整合素连接激酶(ILK)的mRNA表达;细胞免疫荧光检测上皮钙黏素(E-cadherin)、α-平滑肌肌动蛋白(α-SMA)的表达;ELISA法检测纤维连接蛋白(FN)的蛋白水平。结果 C组ILK mRNA,α-SMA、FN蛋白较其他各组表达明显增高(P<0.05),E-cadherin蛋白表达明显下降(P<0.05),而TGF-β1mRNA表达高于除G组以外其余各组(P<0.05)。与G组相比,C组TGF-β1mRNA表达差异无统计学意义(P>0.05),而ILK mRNA,α-SMA、FN蛋白表达明显下降(P<0.05),E-cadherin表达明显升高(P<0.05)。D、E、F组TGF-β1、ILKmRNA、α-SMA,FN蛋白表达逐渐下降,组间两两比较差异有统计学意义(P<0.05);E-cadherin的表达逐渐增高(P<0.05)。结论一定浓度的清蛋白可以诱导体外培养的HK-2细胞发生转分化;螺内酯可抑制人清蛋白诱导HK-2细胞转分化作用,且呈剂量依赖性;TGF-β1/Smads信号通路可能是此过程中的重要通路。
Objective To discuss the effects of spironolactone（SPI） on albumin（Alb） induced transdifferentiation of human kid- ney proximal renal tubular epithelial cells（HK 2 cells）, and whether the process through TGF-β1/Smads signaling pathways. Meth- ods In vitro HK-2 cells were randomly divided into the following seven groups：group A （ without stimulus ）,group B （ 10^- 7 mol/L SPI ）,group C （ 5 mg/mL Alb ）,group D （ 5 mg/mL Alb ＋ 10 8 mol/L SPI）,group E （ 5 mg/mL Alh ＋ 10^- 7 mol/L SPI） ,group F （ 5 mg/mL AIb ＋ 10^-6 moi/L SPI） ,group G （10 tlmol/L TGF-β1 antagonist） 30 min later adding 5mg/mL Alb）. Using RT-PCR to detect TGF-β1 and ILK mRNA expressions; Immunofluorescence detection in E-cadherin and α-SMA proteins ex- pression; ELISA assay for detection of FN protein levels. Results In group C,compared with other groups,ILK mRNA and α- SMA,FN proteins expression were higher than other groups （P〈 0.05）, but E-eadherin protein express declined obviously （P〈0.05） ,at the same time,TGF-β1 mRNA expression was higher than other groups except group G（P〈0.05）. Compared with the group G,TGF-β1 mRNA expression of group C was no obviously difference（P〉0.05） ,but ILK mRNA and α-SMA,E-cadherin,FN proteins expression were distinctly decreased（P〈0.05）. In group D, E, F, TGF-β1, ILK mRNA and α-SMA, FN proteins expres- sions had descended gradually（P〈0.05）. Compared with any two of them, these expressions had obviously decreased （P〈0.05）. On the contrary, E-cadherin expression was in the opposite direction（P〈0.05）. Conclusion A certain concentration of albumin can be induced transdifferentiation of HK-2 cells in vitro. Spironolactone inhibits this process in a dose-dependent manner. TGF-β1 Smads signaling pathway may be play an important role in this process.
出处
《重庆医学》
CAS
CSCD
北大核心
2012年第18期1799-1801,1804,共4页
Chongqing medicine
基金
江西省卫生厅科技计划(20121077)
关键词
螺内酯
肾小管
上皮细胞
细胞分化
转化生长因子Β
spironolactone kidney tubules
epithelial cells
cell differentiation
transforming growth factor beta
