摘要
目的:探讨丹皮酚(Paeonolum,Pae)对TGF-β_1诱导HK-2细胞转分化的抑制作用及机制。方法:将细胞分为5组:无血清对照组、TGF-β_1组、TGF-β_1+10 mg/L Pae组、TGF-β_1+50 mg/L Pae组、TGF-β_1+100 mg/L Pae组。TGF-β_1作用24 h,后加入相应药物继续培养48 h,用倒置显微镜观察各组细胞形态学变化,Western blot法检测E-cadherin和Vimentin蛋白表达的变化,RT-PCR法检测Twist mRNA表达变化。结果:①倒置显微镜下观察:10μg/L TGF-β_1刺激72 h后,HK-2细胞形态发生明显改变,变成长梭形,且细胞间的缝隙变大,Pae各个剂量组作用后,细胞的形态趋向于铺路石状,高剂量组变化最显著。②Western blot结果显示:丹皮酚组能剂量依赖地提高E-cadherin蛋白表达,降低Vimentin蛋白表达。③RT-PCR结果显示丹皮酚能剂量依赖地降低HK-2细胞的Twist mRNA表达。结论:丹皮酚能显著抑制TGF-β_1诱导HK-2细胞转分化萁作用机制可能与抑制Twist mRNA表达有关。
Objective： To explore the effects and mechanism of Paeonolum on TGF- β1- induced epithelial mesenchymal transition in HK- 2 cells. Methods： HK- 2 cells were divided into five groups as follows： control group,TGF- β1 group,TGF- β1＋ 10 mg / L pae,TGF- β1＋ 50 mg / L pae,TGF- β1＋ 100 mg / L pae groups.Using inverted microscope to observe the morphological changes of cells in every group; expressions of E- cadherin and Vimentin protein were detected by Western blot method; expression of Twist mRNA was detected by RT- PCR. Results： 1The cells morphology were changed into fibroblast with TGF- β1（ 10 μg/L） under inverted microscope,and Paeonolum can maintain the morphological stability of HK- 2 cell. 2Western blot result showed that Paeonolum could raise the expression of E- cadherin and decrease the expression of Vimentin. 3RT- PCR result showed that Paeonolum could inhibit the expression of Twist mRNA on HK- 2 cells. Conclusion： Paeonolum can inhibit the renal interstitial fibrosis in vitro,possibly by inhibiting the expression of Twist mRNA.
出处
《中国中医药科技》
CAS
2016年第3期282-284,共3页
Chinese Journal of Traditional Medical Science and Technology
基金
浙江省自然科学基金项目(LQ16H280002)
浙江省中医药管理局项目(2015ZA073)
关键词
丹皮酚
@HK-2细胞转分化
基因表达
Paeonolum
epithelial mesenchymal transition in Hk-2cells
gene expression
