The Effects Of All-trans Retinoic Acid On TGF-?1/Notch Signaling Pathway In Injured Podocytes Induced By Adriamycin

Part I The expression of TGF-?1/Notch signaling pathway and its role in injured podocytes induced by adriamycinObjective To explore the expression of TGF-?1/Notch signaling pathway and its role in injured podocytes induced by adriamycin,by detecting the change of transforming growth factor-?1(TGF-?1),podocin,Notch1 and Jagged1 expressions under adriamycin injury in vitro.Methods Mouse podocyte clone 5(MPC5)cultured in vitro were divided into normal group and model group.Podocytes in normal group were grown in normal RPMI-1640 medium until the end,meanwhile podocytes in model group were subjected to RPMI-1640 medium which including 0.1?g/ml adriamycin for24 hours to construct injured podocytes model.Morphological changes were observed by light microscope.Expression of TGF-?1,podocin,Notch1,Jagged1 m RNA and protein were evaluated by real-time polymerase chain reaction(RTPCR)and western blot.Results(1)The cell morphology of podocytes in normal group were regular,that is,plump,fusiform and arrangement orderly,while it appeared atrophy or hypertrophy,fuzzy boundary,arrange disorderly and increased cellular debris in model group.(2)Compared with normal group,the level of m RNA and protein of TGF-?1,Notch1,Jagged1 increased significantly in model group(all P<0.05),yet the expression of podocin showed an obvious tendency of decrease(all P<0.05).(3)The m RNA expression of Notch1 and Jagged1 were positively correlated with m RNA level of TGF-?1 in model group(r=0.84,r=1,all P<0.05),and negatively correlated with m RNA level of podocin(r=-0.95,r=-0.94,all P<0.05).Conclusion In injured podocytes induced by adriamycin,the expression of TGF-?1/Notch signaling pathway was dramatically enhanced,and TGF-?1/Notch signaling pathway might be involved in podocyte damage.Part II Effects of all-trans retinoic acid on TGF-?1/Notch signaling pathway in podocytesObjective To observe the effects of all-trans retinoic acid(ATRA)on TGF-?1/Notch signaling pathway in injured podocyte induced by adriamycin(ADR)in vitro.Methods Podocytes cultured in vitro were randomly divided into four groups: normal group,model group,ATRA treatment control group and ATRA intervention group.Morphological changes were observed by using optical microscopy.The expressions of TGF-?1,podocin,Notch1,Jagged1 m RNA were evaluated by real-time polymerase chain reaction(RT-PCR)and the corresponding proteins by western blot.Results(1)No obvious changes between normal group and ATRA treatment control group were revealed,in which the plump podocytes and distinct outline were found in light microscope.While podocytes in model group showed disordered arrangement,fuzzy boundary,atrophy,hypertrophy and increased cellular debris.Of note,the podocytes in ATRA intervention group almost returned to normal.(2)In contrast with these in model group,the amounts of TGF-?1,Notch1,Jagged1 were reduced in ATRA intervention group,whereas the level of podocin increased(all P<0.05).(3)There were no significant differences displayed in the transcription and translation levels of the TGF-?1,Notch1 and Jagged1 between normal group and ATRA treatment control group,(all P>0.05).Conclusion ATRA might alleviate podocytes injury through cutting the expressions of TGF-?1,Notch1,Jagged1 and enhancing the expression of podocin in injured podocytes induced by adriamycin.