Prevention of renal ischemia and reperfusion injury by penehyclidine hydrochloride through autophagy activation.

Abstract

Penehyclidine hydrochloride (PHC) suppresses renal ischemia and reperfusion (I/R) injury (IRI); however, the underlying mechanism of action that achieves this function remains largely unknown. The present study aimed to investigate the potential role of autophagy in PHC-induced suppression of renal IRI, as well as the involvement of cell proliferation and apoptosis. A rat IRI model and a cellular hypoxia/oxygenation (H/R) model were established; PHC, 3-methyladenine (3-MA) and rapamycin (Rapa) were administered to the IRI model rats prior to I/R induction and to H/R cells following reperfusion. Serum creatinine was measured using a biochemistry analyzer, whereas aspartate aminotransferase (ASAT) and alanine aminotransferase (ALAT) expression levels were detected using ELISA kits. Renal tissue injury was evaluated by histological examination. In addition, microtubule-associated protein light chain 3B (LC3B) expression, autophagosome formation, cell proliferation and apoptosis were detected in the cellular H/R model. The results demonstrated that I/R induced renal injury in IRI model rats, upregulated serum creatinine, ALAT and ASAT expression levels, and increased autophagic processes. In contrast, pretreatment with PHC or Rapa significantly prevented these I/R-induced changes, whereas the administration of 3-MA enhanced I/R-induced injuries through suppressing autophagy. PHC and Rapa increased LC3B and Beclin-1 expression levels, but decreased sequestome 1 (p62) expression in the cellular H/R model, whereas 3-MA prevented these PHC-induced changes. PHC and Rapa promoted proliferation and autophagy in the cellular H/R model; these effects were accompanied by increased expression levels of LC3B and Beclin-1, and reduced p62 expression levels, whereas these levels were inhibited by 3-MA. Furthermore, PHC and Rapa inhibited apoptosis in the cellular H/R model through increasing Bcl-2 expression levels, and suppressing Bax and caspase-3 expression levels; the opposite effect was induced by 3-MA. In conclusion, PHC suppressed renal IRI through the induction of autophagy, which in turn promoted proliferation and suppressed apoptosis in renal cells.