Necrostatin-1 protects against ischemia/reperfusion injury by inhibiting receptor-interacting protein 1 in a rat flap model

Abstract

The failure of reconstructive surgeries remains a challenge for plastic surgeons. Ischemia reperfusion (I/R) injury is considered to be one of the major problems in flap surgery. Necroptosis is a recently discovered and caspase-3-independent programed necrosis. Necrostatin-1 (Nec-1) is a specific inhibitor of necroptosis. Reports indicate that Nec-1 provides protection in ischemic models, such as brain, kidney, and heart. The aim of this study is to investigate the influence of Nec-1 on the I/R process in rat abdominal skin flaps. Methods: Twenty male Sprague-Dawley rats, weighing 280-320 g, were randomly divided into three groups. The extended epigastric skin flap (6 cm × 9 cm) of rats was used. Three hours of complete ischemia was performed using a clamp, and the clamp was then removed to reperfusion the flap. Twenty-four hours after the onset of the reperfusion, the rats were assessed for flap survival and perfusion analysis. One sample (1 cm × 1 cm) was taken for H&E, TUNEL, electron microscopy, IHC staining for RIP-1, and ELISA analysis for caspase-3 activity. Results: Compared to the CTL group, the flap in the Nec-1 group showed a higher survival rate and better blood perfusion. In histological observation, skin flap in the Nec-1 group showed less inflammatory infiltration than the CTL group. The AI in the CTL group was higher than that in the Nec-1 group and showed typical morphological changes of apoptotic cells. In IHC study, RIP-1 expression was higher in the CTL group. But there was no significant difference between the two groups in caspase-3 activity detection.

Cyclic helix B peptide promotes random‐pattern skin flap survival via TFE3‐mediated enhancement of autophagy and reduction of ROS levels

“…This newly identified programmed necrosis is called ‘necroptosis’ (Vandenabeele et al, 2010). Furthermore, some studies have detected necroptosis in the skin flaps in agreement with the data of RNA sequencing analysis obtained in the present study (Geng et al, 2020; Liu, Zhang, Dong, et al, 2019; Liu, Zhang, Liu, et al, 2019). Thus, we measured the levels of necroptosis by western blotting, immunohistochemistry and immunofluorescence.…”

Evidence of necroptosis in osteoarthritic disease: investigation of blunt mechanical impact as possible trigger in regulated necrosis

“…Degterev et al identified Necrostatin 1 (Nec-1) as a reliable small-molecule inhibitor of necroptotic cell death 7,13 . Moreover, Nec-1 has additionally been found to prevent RIP1-dependent apoptosis and autophagy after traumatic injury 14,15 .…”

The Effect of Necrostatin -1 and Enoxaparin Molecules on Random Pattern Flap Viability

“…Receptor interacting protein kinase-1 (RIPK-1) and receptor interacting protein kinase-3 (RIPK-3) activity is required for TNFR activation (Degterev et al, 2005;Xie et al, 2013). In recent studies, necroptotic cells have been shown in ischaemicreperfusion created in rat skin flap models and it has been reported that necroptosis could have a role in flap necrosis (Liu et al, 2019a). Due to this role of RIPK-1 and RIPK-3 in necroptosis, it has been considered that inhibition of these enzymes could be a good strategy Black Sea Journal of Health Science against necroptosis.…”