Recombinant Human Kallistatin Repairs Cell Injury Induced By Contact Lens Care Solution

In recent years,most literature reports versatile contact lens solution may hurts cornea.It is reported that,versatile contact lens solution may cause corneal epithelium oxidative stress response and inflammatory reaction which will cause cell apoptosis,and versatile contact lens solution cause cell inflammatory reaction through activate NF-?B signaling pathway.Animal experiments also show that versatile contact lens solution causes obvious rabbit cornea tissue damage and reduce resistance to bacteria of cornea.Kallistatin?Kal?is a kind of serine protease inhibitor with pleiotropic effects it can involve in regulating multiple cellular pathways,facilitate vascular repair,reduce hypertension,inflammation,oxidative stress,cell apoptosis,etc.,and it can inhibit the NF-?B signaling pathways,plays the role of anti-inflammatory.SERPINA3 K,the family of Kal has anti-inflammatory,anti-angiogenesis and antioxidant activity on HCE cell.Base on Kal pleiotropy and SERPINA3K's biological activity on the corneal epithelium cells,we surmise that Kal may have a good repair effect on damage of HCE.Major research contents:?1?Preparation of Kal albumen: To gain high expression strain through screening recombinant human Kal?rhKal?yeast strains built in laboratory,do inducible expression with fermentation technology,to further separate and purify rhKal protein,SDS-PAGE detection and Quantities ELISA rhKal with high purity and high concentration.?2?The five kinds of commercially available contact lens care solution?including antiseptic except hydrogen peroxide?toxicity verify on HCE cell: with 3 h effect time,with the CCK-8 method screening concentration of care solution make HCE vigor declined significantly and record its concentration,then observe the cell morphology under this concentration,detect levels of cell super oxide anion and cell apoptosis.?3?RhKal's repair effect to HCE damage caused by care solution: applying rhKal protein to five cell damage models caused by care solution,to study the biological activity of oxidation resistance,resistance to apoptosis;?4?Repair effect of RhKal on injury of HCE and human conjunctival epithelial cells?HConEpiC?caused by H₂O₂: with 3 h effect time,with H₂O₂ to build damage models of HCE and HConEpi C,study the repair effect of rhKal protein on the damage.The results manifestRhKal has significant repair effect on HCE cell damage caused by five kinds of contact lens care solution,can reduce intracellular superoxide anion,inhibit cell apoptosis caused by care solution,and it has a certain dose dependent.RhKal also has significant repair effect on cell damage of HCE and HConEpiC caused by H₂O₂,can effectively inhibit cell apoptosis.This study shows that rhKal has to repair effect on damage caused by the contact lens care solution,and laid a solid foundation for developing a damage repair fluid for contact lens care solution.