| Objective:To explore the effect of1α,25-dihydroxyvitamin D3on adriamycin-inducedepithelial-mesenchymal transition of podocytes.Methods:Mouse podocytes in vitro cultured under differentiating conditions for10days weredivided into6groups which were the control group(C group),the model group(ADRgroup,0.5ug/ml), the low dose1α,25-dihydroxyvitamin D3group(LVDgroup,1nmol/L), the high dose1α,25-dihydroxyvitamin D3group(HVDgroup,10nmol/L), the valsartan group(ARB group,10umol/L)and the combinationgroup(AHVD group,high dose1α,25-dihydroxyvitamin D310nmol/L,valsartan10umol/L). Each group was continually incubated with drugs for24h and48h. Theproliferation and apoptosis of podocytes in each group was detected by MTT andflow cytometry(FCM) respectively and the expression of protein molecules whichwere related to epithelial-mesenchymal transition such as synaptopodin、P-cadherin、desmin、 FSP-1、TGF-βandβ-catenin was detected by reversetranscription polymerase chain reaction(RT-PCR). In addition, the expression ofprotein molecules such as synaptopodin、P-cadherin、desmin andβ-catenin was alsodetected by Western Blot.Results:1、MTT results showed that after intervening with drugs for24h and48h, theproliferation of podocytes in ADR group was significantly decreased (P <0.01);compared with the ADR group, the proliferation of podocytes was increasedobviously(P＜0.01)in the LVD group, HVD group, ARB group and AHVD group.2、After continually incubated with drugs for24h and48h, the expression of synaptopodin mRNA and protein were decreased in ADR group(P <0.01). Comparedwith ADR group in24h, the expression of synaptopodin mRNA was increasedsignificantly(P <0.01) in ARB group, LVD group, HVD group and AHVD group,there was no significant difference between the four drug intervention group. Inaddition, the expression of synaptopodin mRNA and protein were increased (P <0.05)in LVD group and HVD group when compared with ADR group in48h. Butcompared with ADR group in24h and48h, the expression of synaptopodin proteinwas decreased significantly in ARB group (P <0.01). When it comes to P-cadherin,the expression of P-cadherin mRNA and protein were decreased in ADR group(P<0.01) after intervening with drugs for24h and48h; then compared with ADR group,the expression of P-cadherin mRNA was increased (P <0.05) in ARB group, LVDgroup and HVD group. But at protein levels in24h, the AHVD group does not havesignificant difference (P>0.05) when compared with ADR group.3、The expression of desmin mRNA and protein were increased in ADR group(P<0.01)after intervening with drugs for24h and48h; then compared with ADR group,the expression of desmin mRNA and protein were decreased (P <0.01) in ARB group,LVD group, HVD group and AHVD group. When it comes to FSP-1, the expressionof FSP-1mRNA was increased (P <0.01) in ADR group when compared with controlgroup in24h; then compared with ADR group, the expression of FSP-1mRNA wasdecreased (P <0.01) in ARB group, LVD group, HVD group and AHVD group. Inaddition, compared with control group in48h, the expression of FSP-1mRNA wasincreased slightly in ADR group, but had no statistical significance; moreover, theexpression of FSP-1mRNA was decreased (P <0.05) in LVD group, HVD group andAHVD group when compared with ADR group.4、After continually incubated with drugs for24h and48h, the expression of β-catenin mRNA and protein were increased in ADR group(P <0.01); then comparedwith ADR group, the expression of β-catenin mRNA and protein were decreased (P<0.05) in ARB group, LVD group, HVD group and AHVD group. In our study, theexpression of TGF-βmRNA was increased (P <0.01) in ADR group when compared with control group in24h; then compared with ADR group, the expression of TGF-βmRNA was decreased significantly(P <0.01) in ARB group, LVD group, HVD groupand AHVD group. Moreover, it’s showed that the expression of TGF-βmRNA wasincreased (P <0.01) in ADR group when compared with control group in48h; thencompared with ADR group, the expression of TGF-β mRNA was decreasedsignificantly(P <0.01) in HVD group and AHVD group, the other two groups had nosignificant difference.Conclusion:1、Adriamycin can induce podocytes occur EMT, then establish EMT model ofmouse podocytes in vitro.2、1α,25-dihydroxyvitamin D3can antagonize podocyte injury by repairingsynaptopodin, P-cadherin expression and inhibiting desmin,FSP-1expression. Itstates that1α,25-dihydroxyvitamin D3can protect podocytes by inhibiting EMT inpodocytes and it has not dose-dependent and time-dependent manner.3、In this study, our experimental dose of valsartan combines with high dose1α,25-dihydroxyvitamin D3on the inhibition of podocyte EMT has not synergisticeffect.4、Our study confirms that1α,25-dihydroxyvitamin D3suppresses podocytesoccurring EMT by inhibiting the expression ofβ-catenin and TGF-βwhich aresignal transduction factors of podocytes EMT signal transduction pathway. |
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