Everolimus Alleviates Renal Allograft Interstitial Fibrosis by Inhibiting Epithelial-to-Mesenchymal Transition Not Only via Inducing Autophagy but Also via Stabilizing IκB-α

Chronic allograft dysfunction (CAD) is the major cause of late graft loss in long-term renal transplantation. In our previous study, we found that epithelial-mesenchymal transition (EMT) is a significant event in the progression of renal allograft tubulointerstitial fibrosis, and impaired autophagic flux plays a critical role in renal allograft fibrosis. Everolimus (EVR) has been reported to be widely used to prevent the progression of organ fibrosis and graft rejection. However, the pharmacological mechanism of EVR in kidney transplantation remains to be determined. We used CAD rat model and the human kidney 2 (HK2) cell line treated with tumor necrosis factor-alpha (TNF-alpha) and EVR to examine the role of EVR on TNF-alpha-induced EMT and transplanted renal interstitial fibrosis. Here, we found that EVR could attenuate the progression of EMT and renal allograft interstitial fibrosis, and also activate autophagy in vivo. To explore the mechanism behind it, we detected the relationship among EVR, autophagy level, and TNF-alpha-induced EMT in HK2 cells. Our results showed that autophagy was upregulated upon mTOR pathway inhibition by EVR, which could significantly reduce expression of TNF-alpha-induced EMT. However, the inhibition of EVR on TNF-alpha-induced EMT was partly reversed following the addition of autophagy inhibitor chloroquine. In addition, we found that TNF-alpha activated EMT through protein kinase B (Akt) as well as nuclear factor kappa B (NF-kappa B) pathway according to the RNA sequencing, and EVR's effect on the EMT was only associated with I kappa B-alpha stabilization instead of the Akt pathway. Together, our findings suggest that EVR may retard impaired autophagic flux and block NF-kappa B pathway activation, and thereby prevent progression of TNF-alpha-induced EMT and renal allograft interstitial fibrosis.

Automated summary

This study found that everolimus can attenuate the progression of renal allograft interstitial fibrosis and activate autophagy in vivo. It was observed that everolimus reduces expression of TNF-alpha-induced EMT through upregulation of autophagy. In addition, everolimus affects EMT by stabilizing I kappa B-alpha instead of the Akt pathway.