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In the co-culture system, MCECs derived from the model group exhibited significant reduced ZO-1 and Occludin expression compared to those from the Control group, whereas GHK-Cu elevated theses protein levels (Figures 4GI)

Detection of protein carbonylation Levels of carbonylated proteins were determined with the OxyBlot protein oxidation detection kit following the manufacturers protocol (Millipore, Billerica, MA, USA)

The membranes were washed using TBST 3 times, 8 min each time

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This reaction is termed the Fenton reaction (Valko et al