摘要
目的研究转化生长因子β1(transforming growth factor β1,TGFβ1)对人肾小管上皮细胞株(human kidney proximal tubular epithelial cell line,HKCs)表达糖原合成酶激酶3β(glycogen synthase kinase-3β,GSK-3β)的影响。方法将体外培养的的HKCs随机分为4组:正常对照组(C组):无血清培养基(free serum medium,FSM)培养;Ta组、Tb组和Tc组:用含不同浓度TGFβ1(Ta组:5ng/ml,Tb组:10ng/ml,Tc组:20ng/ml)的FSM培养。12h后,分别用RT-PCR、Western blot检测HKCs GSK-3β的mRNA、总蛋白表达量以及蛋白磷酸化水平。结果(1)GSK-3β mRNA和总蛋白在各组间表达量差异无统计学意义(P>0.05),和C组相比各T组GSK-3β蛋白磷酸化水平显著升高(P<0.01);(2)Tb组与Tc组相比,GSK-3β磷酸化水平更高(P<0.01);(3)Tb组与Tc组相比,GSK-3β磷酸化水平差异无统计学意义(P>0.05)。结论TGFβ1虽不影响HKCs的GSK-3β mR-NA和总蛋白表达量,但能提高GSK-3β蛋白磷酸化水平,后者可能参与TGFβ1诱导的HKCs转分化过程。
Objective To investigate the effects of transforming growth factor β1 （TGFβ1）on the expression of glycogen synthase kinase-3β（GSK-3β） in human kidney proximal tubular epithelial cell line（HKCs）. Methods HKCs cultured in vitro were divided into four groups at random：control （C） group： the HKCs were cultured with free serum medium（FSM） ; Ta, Tb and Tc groups： the HKCs were cultured with FSM containing different concentrations of TGFβ1 （Ta ： 5ng/ml, Tb：10ng/ml, Tc ： 20ng/ml） respectively. After treatment for 24h,the expressions of GSK-3β mRNA, protein and phospho-GSK-3β （Ser 9） were examined by reverse transcription-polymerase chain reaction （RT-PCR） and Western blot. Results （1） There were no significant difference among the groups in the levels of GSK-3β mRNA and protein（P〉0.05） ; but T groups statistically increased than C group（P〈0. 01） ;（2）Tb group had more significant phosphorylation of GSK-3β than Ta group（ P〈0.01 ） ; （3）There was not visible difference of phosphorylated GSK-3β between Tb group and Tc group. Conclusion Although the treatment of TGFβ1 has no visible effect on the expression of GSK-3β mRNA and protein, it may increase the level of phosphorylated GSK-3β when the concentrations of TGFβ1 raising. It may be one of the mechanisms that TGFβ1 induces TEMT in HKCs.
出处
《重庆医学》
CAS
CSCD
2007年第5期414-416,共3页
Chongqing medicine
基金
重庆市自然科学基金资助课题(2006BB5096)
关键词
肾小管上皮细胞
转化生长因子Β1
糖原合成酶激酶3Β
human kidney proximal tubular epithelial cell line
transforming growth factor-β1
glycogen synthase kinase-3β
