Low-dose Cold Atmospheric Plasma Promotes Schwann Cells Proliferation By Activating PI3K/Akt/mTOR Pathway

Objective: The treatment of peripheral nerve injury(PNI)is not only a difficult problem in medicine,but also a great demand of social and economic development in China.At present,the treatment and research of PNI repair still mainly stay at the suture technology level,and the actual clinical treatment effect for nerve injury patients is often not satisfactory.Schwann cells(SCs),as important cells to maintain the stability of normal nerve internal environment and promote regeneration after PNI,play an important role in the repair of PNI.Cold atmospheric plasma(CAP)is a new method for biomedical treatment.Studies have shown that low-dose CAP can promote cell proliferation.In this study,low-dose CAP was used to treat SCs to observe the effects of CAP on SCs and to study its related mechanisms.Methods: In this study,plasma generated by dielectric barrier discharge(DBD)was used to treat SCs cultured in vitro,and the treatment time was 0s,7s,14 s,21s,and 28 s,respectively.We used ROS and RNS detection kits to detect ROS and RNS concentrations in cell culture medium.Intracellular ROS and RNS were detected by DCFH-DA probe and DAF-FMDA probe.CCK-8 was used to detect the effect of plasma on SCs proliferation.Cell cycle distribution in each group was detected by flow cytometry.Western Blot was used to detect the expression of PI3K/Akt/m TOR pathway related proteins and cell cycle related proteins.Results: The results of intracellular and extracellular ROS and RNS showed that the amount of ROS and RNS was always positively correlated with the plasma treatment time,both inside and outside the cells.CCK-8 results showed that low-dose CAP could promote the proliferation of SCs,while high-dose CAP could inhibit the growth of SCs.The results of cell cycle detection showed that appropriate CAP treatment could promote the transformation of cell cycle from G0/G1 phase to S+G2/M phase,thus promoting the proliferation of SCs.Western Blot results showed that low-dose CAP may promote the proliferation of SCs through ROS activation of PI3K/Akt/m TOR pathway and cell cycle related pathways.Conclusion: Low-dose CAP can promote the proliferation of SCs.Low-dose CAP may promote the proliferation of SCs through ROS activation of PI3K/Akt/m TOR pathway and cell cycle related pathways.