Ischemic postconditioning inhibits apoptosis in an in vitro proximal tubular cell model

Weng,Xiaodong; Wang,Lei; Chen,Hui; Liu,Xiuheng; Qiu,Tao; Chen,Zhiyuan

doi:10.3892/mmr.2015.3344

Ischemic postconditioning inhibits apoptosis in an in vitro proximal tubular cell model

Authors:
- Xiaodong Weng
- Lei Wang
- Hui Chen
- Xiuheng Liu
- Tao Qiu
- Zhiyuan Chen
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Affiliations: Department of Urology, Renmin Hospital of Wuhan University, Wuhan University, Wuhan, Hubei 430060, P.R. China
Published online on: February 12, 2015 https://doi.org/10.3892/mmr.2015.3344
Pages: 99-104
Copyright: © Weng et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

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Abstract

Ischemia‑reperfusion is a common injury of clinical ischemic disease and surgical lesions. Ischemic postconditioning (IPO) improves the ability of organs subjected to ischemia to tolerate injury. However, renal IPO studies have been based on animal models. In order to gain insights into IPO‑induced alterations at the cellular level, an in vitro model for IPO was designed using the rat proximal tubular cell line NRK‑52E. This model was established by placing NRK‑52E cells in ischemic conditions for 3 h, then exposing cells to three cycles of reperfusion for 10 min and finally to ischemic conditions for 10 min (postconditioning). The cells were cultured further in reperfusion conditions for 3, 6, 12 and 24 h. Flow cytometry and Hoechst were used to assess apoptosis. The protein expression of B‑cell lymphoma 2 (Bcl‑2), Bcl‑2‑associated X protein (Bax), caspase‑3, cleaved caspase‑3 and caspase‑8 were analyzed by western blotting. The results demonstrated that apoptosis occurred in cells subjected to ischemia/reperfusion (I/R) alone or with postconditioning following reperfusion for 24 h. Cells subjected to I/R demonstrated increased expression of Bax, cleaved caspase‑3 and caspase‑8 at the end of reperfusion. However, the levels of Bax, cleaved caspase‑3 and caspase‑8 were significantly attenuated in cells, which had undergone IPO. In conclusion, apoptosis was observed in cells subjected to 3 h of ischemia‑reperfusion injury and IPO was able to inhibit this apoptosis. IPO inhibited apoptosis by inhibiting the caspase pathway thereby exerting protective effects.

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