Effect of hydrogen-rich saline on TNF-α, NF-κB and apoptosis in skin flap after ischemia/reperfusion injury

Abstract

Objective To study the protective effect of hydrogen-rich saline (HRS) on apoptosis in skin flap after ischemia/reperfusion injury.Methods Total 18 Sprague-Dawley rats were randomly divided into three groups:a HRS treated group and two physiological saline treated groups (controls 1,2).The rats were anesthetized and an extended abdominal skin flap (6 cm × 9 cm) was elevated in each animal.Ischemia was induced by clamping the left right pedicle for 3 h,then HRS was administered intraperitoneally 10 min before reperfusion,and physiological saline was injected in control groups 1 and 2.In the control group 2,the flaps were elevated without occluding the artery and vein.Five days postoperation,apoptosis,TNF-α level in flap were measured with ELISA,NF-κB in nucleus was determined by Westernblot.Results Apoptotic rate represented (39.72±8.09) ％in HRS group and (69.43±13.27) ％ in control group 1,respectively.Treatment with HRS resulted in a marked reduction in apoptotic rate.TNF-α level was (516.408±38.674) pg/ml in the control group 1,a significant reduced TNF-α was measured in HRS group,accounting for (269.136 ±24.530) pg/ml.Moreover,NF-κB activation was significantly down-regulated by HRS.In control group 2,no significant apoptosis was observed because of non-blood occlusion,and there was no marked elevation of TNF-α and NF-κB.Conclusions HRS can protect skin from ischemia/reperfusion injury,attenuate apoptosis in flaps,which may be associated with the inhibition of TNF-α and NF-kb elevation. Key words: Hydrogen-rich saline(HRS); Skin flap; Apoptosis; TNF-κB; TNF-α