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a

b c
Figure 1: The evaluation of the abdominal skin flaps 72 h following ischemia‑reperfusion. Black zones represent the necrotic areas. Red, yellow, and the
adjacent blue areas represent surviving areas with rich blood perfusion. (a) Representative photographs of abdominal skin flap microcirculation in the five
groups are shown; (b) the survival rate of the total flap area. Flap survival rates were markedly higher in the hyperbaric oxygen, hydrogen‑rich saline, and
hyperbaric oxygen + hydrogen‑rich saline groups; (c) the average blood perfusion of total skin flaps. The hyperbaric oxygen + hydrogen‑rich saline group has
the greatest blood perfusion among the surgery groups. IR: ischemia reperfusion, HBO: postoperative hyperbaric oxygen, HRS: hydrogen‑rich saline

Apoptotic index evaluation HBO + HRS (0.36 ± 0.17) groups IR (P < 0.001).
Apoptotic cell death was observed with TUNEL Significant differences between the HBO + HRS group and
staining [Figure 2a]. The number of apoptotic cells increased the HBO and HRS groups were also found (HBO + HRS vs.
in the IR group (45.85% ± 6.64%) as compared with the HRS, P < 0.05; HBO + HRS vs. HBO, P < 0.01). However,
sham group (8.57% ± 4.12%). Apoptotic cell number there was no statistical difference between the HBO and
was reduced with HBO preconditioning, HRS and HRS HRS groups [Figure 3].
and HBO preconditioning used cooperatively. HRS and
HBO preconditioning used cooperatively were more Western blot for pASK1 and Bcl‑2/Bax
efficient in reducing cell apoptotic death than that of The protein expression of pASK1, Bcl‑2 and Bax proteins
HBO preconditioning or HRS used independently. The was analyzed by Western blot [Figure 4a]. pASK1
AI in HRS was 32.69% ± 6.80%, in the HBO group was expressed the highest level in the IR group (0.25 ± 0.04)
30.80% ± 7.13%, and in the HBO + HRS group was compared with other groups and was significantly
20.24% ± 6.90%. There were significant differences decreased in the HBO + HRS group (0.13 ± 0.05) as
between each surgical group, (IR vs. HBO, P < 0.001; IR vs. compared to the IR (0.25 ± 0.04), HRS (0.17 ± 0.04) and
HRS, P < 0.001; IR vs. HBO + HRS, P < 0.001; HBO vs. HBO (0.18 ± 0.03) groups. Statistical differences were
HBO + HRS, P < 0.01; and HRS vs. HBO + HRS, P < 0.001) observed in HBO + HRS vs. IR, P < 0.001; HRS vs. IR,
with the exception of HBO versus HRS [Figure 2b]. P < 0.001; HBO vs. IR, P < 0.001; HBO + HRS vs. HBO,
P < 0.05; and HBO + HRS vs. HRS, P < 0.05 [Figure 4b].
Caspase‑3 activity
Compared to the IR group (1.25 ± 0.26), the The ratio between the level of Bcl‑2 to Bax expression
caspase‑3 relative activity was significantly lower increased in the HBO (2.06 ± 0.49), HRS (2.90 ± 0.65)
in the HBO (0.59 ± 0.12), HRS (0.53 ± 0.15), and and HBO + HRS (3.27 ± 0.42) groups. The Bcl‑2/Bax

Plast Aesthet Res || Vol 2 || Issue 6 || Nov 12, 2015 335