Department of Orthopedic Surgery, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou Medical University, Wenzhou, Zhejiang, China.
Department of Hand and Plastic Surgery, The Second affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, The Second School of Medicine, Wenzhou Medical University, Wenzhou, Zhejiang, China.
J Surg Res. 2020 Nov;255:575-582. doi: 10.1016/j.jss.2020.03.044. Epub 2020 Jul 7.
Necrosis of the perforator flap is a critical problem. Fasudil, an inhibitor of Rho-associated coiled-coil containing kinase, has antiapoptosis activity and attenuates oxidative stress in many diseases. We characterized the effects of fasudil through intraperitoneal injection on perforator flap survival and identified its possible mechanism.
Rats were divided into a control group (without surgery), a flap group (only surgery), and a fasudil group (surgery plus fasudil). Perforator flaps were made on the backs of the rats. The expression of vascular endothelial growth factor, the protein kinase B (PKB/Akt), endothelial nitric oxide synthase, Bax, Bcl-2, Beclin-1, P62, and LC3 II/LC3 I was determined by Western blot at day 3 after surgery. Nitric oxide (NO) components, superoxide dismutase, and malondialdehyde were also measured at day 3. The survival rate and laser Doppler perfusion imaging were performed at day 7 after surgery.
The group with fasudil treatment exhibited the higher survival rates and angiogenesis levels. Fasudil also induced the activation of Akt/eNOS/NO pathway detected by the Western blot and NO expression kit. Furthermore, Western blot results showed fasudil-attenuated apoptosis through a raised Bcl-2/Bax rate and enhanced autophagy levels through raised beclin-1, decreased p62, and the elevated rate of LC3 II/LC3 I. Finally, fasudil increased superoxide dismutase and decreased malondialdehyde.
In conclusion, fasudil treatment decreased necrosis of perforator flaps possibly by affecting the Akt/eNOS/NO pathway, attenuating apoptosis and activating autophagy.
穿支皮瓣的坏死是一个关键问题。法舒地尔是一种 Rho 相关卷曲螺旋激酶抑制剂，具有抗细胞凋亡作用，并能减轻许多疾病中的氧化应激。我们通过腹腔注射法舒地尔来研究其对穿支皮瓣存活的影响，并确定其可能的机制。
将大鼠分为对照组（无手术）、皮瓣组（仅手术）和法舒地尔组（手术加法舒地尔）。在大鼠背部制作穿支皮瓣。术后第 3 天，通过 Western blot 检测血管内皮生长因子、蛋白激酶 B（PKB/Akt）、内皮型一氧化氮合酶、Bax、Bcl-2、Beclin-1、P62 和 LC3 II/LC3 I 的表达。还在术后第 3 天测量了一氧化氮（NO）成分、超氧化物歧化酶和丙二醛。术后第 7 天行皮瓣成活率和激光多普勒灌注成像检查。
法舒地尔治疗组的皮瓣成活率和血管生成水平较高。Western blot 和 NO 表达试剂盒检测到，法舒地尔还诱导了 Akt/eNOS/NO 通路的激活。此外，Western blot 结果显示，法舒地尔通过提高 Bcl-2/Bax 比率、增强自噬水平（通过降低 p62 和提高 LC3 II/LC3 I 比率）来减轻皮瓣的凋亡。最后，法舒地尔增加了超氧化物歧化酶，降低了丙二醛。
总之，法舒地尔治疗可能通过影响 Akt/eNOS/NO 通路、减轻凋亡和激活自噬来减少穿支皮瓣的坏死。
