title: [Impact of advanced glycosylation end products-modified human serum albumin on migration of epidermal keratinocytes: an in vitro experiment].
abstract: OBJECTIVE
  To explore the impact of advanced glycosylation end products (AGE)-modified human serum albumin (AGE-HSA) on keratinocyte migration and the mechanism thereof.
  
  
  METHODS
  AGE-HSA was prepared in vitro. Epidermal keratinocytes from Sprague-Dawley rats' back were cultured and treated with AGE-HSA of the terminal concentrations of 0, 30, 60, 90, 120, and 150 microg/ml for 1, 3, 5, and 7 days respectively. MTT method was used to detect the keratinocyte adhesion rate, expressed by absorbance. Keratinocyte migration ability was assessed by scratch wound healing assay and Transwell assay. Expression of integrin alpha3 was determined by flow cytometry. Scanning electron and inverted microscopes were used to observe the pseudopodium and microfilament of the keratinocytes. Immunofluorescence staining was used to detect the form of F-actin in the cells.
  
  
  RESULTS
  The adhesion rates of the keratinocyte cultured with AGE-HSA for 12 and 24 hours were (0.112 +/- 0.022) and (0.173 +/- 0.012) respectively, both significantly lower than those of the control group [(0.122 +/- 0.004) and (0.267 +/- 0.024) respectively, both P < 0.05)]. Scratch wound healing assay showed that the amount of migrating cells in the AGE-HSA group was (7 +/- 4)/HP, significantly less than that of the control group [(61 +/- 11)/HP, P < 0.05)], and Transwell assay showed that the amount of migrating cells in the AGE-HSA group was (72 +/- 18)/HP, significantly less than that of the control group [(288 +/- 52)/HP, P < 0.05]. The expression rate of keratinocyte integrin alpha3 in the AGE-HSA group was (3.2 +/- 1.2)%, significantly lower than that in the control group [(36.6 +/- 11.2)%, P < 0.05]. The spreading of cell body, and the formation of pseudopodium and microfilament of the AGE-HSA group were all depressed in comparison with the control group.
  
  
  CONCLUSIONS
  Keratinocyte migration is inhibited by AGE accumulation in high glucose condition. The mechanism may be the abnormality in the integrin inside-out signaling pathway and AGE-RAGE signaling pathway.
authors.affiliation: First Affiliated Hospital of Guangxi Medical University, Nanning 530021, China.
authors.forename: Zhen-qiang
authors.initials: ZQ
authors.lastname: Song
authors.forename: Run-xiu
authors.initials: RX
authors.lastname: Wang
authors.forename: De-min
authors.initials: DM
authors.lastname: Yu
authors.forename: Peng-hua
authors.initials: PH
authors.lastname: Wang
authors.forename: Shu-liang
authors.initials: SL
authors.lastname: Lu
authors.forename: Ming
authors.initials: M
authors.lastname: Tian
authors.forename: Ting
authors.initials: T
authors.lastname: Xie
authors.forename: Fei
authors.initials: F
authors.lastname: Huang
authors.forename: Guo-zhi
authors.initials: GZ
authors.lastname: Yang
mesh_terms: D000418:Albumins / Q000378:metabolism / Q000494:pharmacology*; D000818:Animals; D002465:Cell Movement; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000166:cytology* / Q000187:drug effects; D000078404:Epidermal Cells; D017127:Glycation End Products, Advanced / Q000494:pharmacology*; D006801:Humans; D015603:Keratinocytes / Q000166:cytology* / Q000187:drug effects; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D012709:Serum Albumin / Q000494:pharmacology*; D000075462:Serum Albumin, Human
chemical_list: D000418:Albumins; D017127:Glycation End Products, Advanced; D012709:Serum Albumin; C109748:advanced glycation end products-human serum albumin; D000075462:Serum Albumin, Human