Yongming Tao*, Jia Wang**,#, Yanhong Liu*, Jianxi Pan***, Zhixiang Ma*, Wenjun Zhen***
*Department of Hand and Foot Surgery, Lanzhou Chinese Medicine Orthopaedics Hospital, Lanzhou, 730030, Gansu Province, China - **Department of Rheumatology, Affiliated Hospital of Gansu University of Traditional Chinese Medicine, Lanzhou 730020, Gansu Province, China - ***Department of Orthopaedics, Lanzhou Hospital of Traditional Chinese Medicine, Lanzhou 730030, Gansu Province, China
Objective: To investigate the protective effect of thioredoxin-1 (Trx-1) on ischemia reperfusion flap and analyze the possible mechanism.
Methods: Using human immortalized keratinocytes (HaCAT cells) as study cells, the optimal CoCl2 concentration and time was used to induce cell hypoxia in vitro. Cells were divided into hypoxia non-intervention group (CoCl2 was added into the cell culture medium to induce cytochemical hypoxia), hypoxia intervention group (CoCl2 was added into the cell culture medium to induce cytochemical hypoxia and rhTrx-1 was added for drug intervention), and negative control group (conventional cell culture medium was added to the cell culture medium).Cell viability was studied by CCK-8 method. Total protein was extracted and its concentration was determined. Expression levels of apoptosis-related proteins were analyzed by Western Blotting.
Results: The concentration of CoCl2 was 80μmol/L, and the cell activity was about 50% of that of the control group when treated with HaCAT cells for 24h, which was used as the concentration of follow-up experiments. After CoCl2 was applied to HaCAT cells for 24h, the expression levels of apoptosis-related proteins ASK1 and c-PARP in the non-hypoxia intervention group were significantly higher than those in the control group, and the expression levels ofTrx-1 were significantly lower than those in the control group (P < 0.05).The expression levels of apoptosis related proteins ASK1 and c-PARP in the hypoxia intervention group were significantly lower than those in the non-hypoxia intervention group, and the expression levels ofTrx-1 were significantly higher than those in the non-hypoxia intervention group (P < 0.05).
Conclusion: Trx-1 preconditioning can reduce the ischemia reperfusion injury of the flap, and its mechanism may be related to the scavenging of reactive oxygen species and the inhibition of ASK-MAPK cell apoptosis signaling pathway, soTrx-1 can be used as the drug intervention target to reduce the ischemia reperfusion injury of the flap.
Thioredoxin-1, Antioxidant, ASK-MAPK, Ischemia-Reperfusion.