| Objective To identify the role of EPO in the C3a induced epithelial mesenchymal transition and the effect to renal fibrosis of unilateral uretreal obstructire rats.Methods1. In vitro experiment: The HK-2 cells were divided into 6 groups namely control group, 10U/ml EPO group, 3ng/mlTGF-βgroup, 3ng/ml TGF-β+10U/ml EPO group, 0.1μM C3a group and 10U/ml EPO + 0.1μM C3a group. The expressions ofα-SMA, E-cadherin and C3 mRNA were investigated by applying RT-PCR, Western Blot and immunofluorescence, respectively.2. In Vivo experiment: The model of UUO rats was established, and all animal subjects were assigned to four groups: sham operation group, UUO control group, Low dose EPO(100U/Kg EPO) treatment group and high dose EPO (1000U/Kg EPO)treatment group. The subjects were given an injection of EPO from 3days after operation to the 14th day. All animal subjects were given methane to induce anesthesia and consequently executed. The pathological change of renal was observed, and immunohistochemistry was performed to measure the expression of SMA, E-cadherin and C3 in the renal of rats.Results1. In vitro experiment: We observed the up-regulated expressions ofα-SMA mRNA and protein in HK-2 cells after intervention of C3a and TGFβ. In the contrast, the expressions of E-cadherin mRNA and protein were down-regulated. Meanwhile, the expression of C3 mRNA and protein was enhanced. However, the effects of C3a and TGFβwere inhibited after the intervention of EPO.2. In vivo experiment: With the treatment of EPO in the UUO rats, the expression of C3 andα-SMA in renal was significantly down-regulated and the up-regulated expression of E-cadherin was also observed.Conclusion1. EPO is capable of suppressing the epithelial mesenchymal transition induced by C3a and TGFβ.2. The enhanced expression of C3 can be observed in UUO rats, and EPO was also capable of inhibit both the expression of C3 and EMT,the inhibition of EPO may be mediated through the expression of C3.
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